ヒト・イヌの悪性腫瘍におけるがん抑制因子Protein Phosphatase 2A再活性化の抗がん効果および分子機構の解明

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(1)Protein Phosphatase 2A. 2020. 3.

(2) Protein Phosphatase2A. 2020. 3.

(3) .............................................................................................. 1 -. Protein phosphatase2A................................................................................... 2. -. PP2A. ................................................................................................... 4. -. ......................................................................................................... 7 perphenazine. ............................................... 8. -. ..................................................................................... 9. -. ....................................................................................................... 10 - -. PPZ. T. .................................................................................................................... 10 - -. PPZ. T. ................................................................................................................ 12 - -. PPZ. T. Akt. ................................................................................................................ 14 - -. PPZ. T. ERK1/2. ........................................................................................................ 17 - -. PPZ. ERK1/2 - -. Akt. PP2A. Akt ......................................................................... 20 T. ................................................................................................................ 24 -. ............................................................................................................... 26.

(4) - -. .................................................. 26. - -. PPZ. .................................................. 27. - -. PPZ. ...... 30. - -. PPZ. ERK1/2. - -. ........................................... 30 .............................................................................. 31. ............................................................................................................... 32 SET. -. ................................................... 33. ............................................................................................. 34 - -. PP2A. - -. SET. - -. SET. - -. .................................................... 34 ................................................... 35. PP2A SET. - -. SET. .................................................... 35 .................................................................. 36. ................................................................................................ 37 ......................................................................................................... 38. - -. SET. - -. SET. ........... 38. ................................................................................................................................ 41 - -. -. SET. ....... 43. - -. ERK1/2. - -. SET. ........ 45 .................. 47. ................................................................................................................. 49.

(5) - -. SET. ........... 49. - -. SET .................................................................................................................... 50. - -. SET. FTY720. cisplatin. ........ 51. ................................................................................................................. 52 SET. PP2A. -. ................................................. 53 ..................................................................................... 54. - -. PP2A/B56. - -. SET. - -. ..................................................................... 54 PP2A. ................................................................ 54. ................................................................................................ 55. -. ......................................................................................................... 56 - -. PP2A/B56. - -. .............................. 56 B56g3. - - - - -. B56g3 B56g3. - -. ............. 59 ............. 62. B56g3. - - -. .............................. 59. ........... 64 ..................................... 69. Xanthohumol. KN-93. ................................................................................................................................ 73 - -. KN-93. ........................ 75. - -. KN-93. .................................... 77.

(6) -. ................................................................................................................. 80 - - -. PP2A. .............................................................. 80. SET. PP2A. - -. B56. - -. Xanthohumol. ............................ 81 ................................................. 83. KN-93. ........................................................................................................................ 84 -. ................................................................................................................. 85 ........................................................................................................ 86. -. PP2A. -. SET. -. PP2A. ........................................... 87 ......................................................................... 90 ................................... 91 ............................................................................ 93. -. ..................................................................................................... 94. -. DNA. ...................... 94. - -. ................................................................................ 94. - -. .... 95. - -. ................................................ 96. - -. ................................ 96. -. ............................................................. 97. -. ......................................... 98 - -. .................................................................................... 98.

(7) - -. pull down assay .............................................................. 99 - -. FLAG M2. .............................................. 99. - -. GST pull down assay ........................................................................... 100. -. ............................................................... 100. -. ....................................................................................... 100. -. NanoBiT. ................................................................................................... 101. - -. NanoBiT. ......................................................... 101. - -. NanoBiT. A549. ................................................... 103. - -. NanoBiT. A549. .................. 103. - -. In vitro NanoBiT. ............................................................................. 104. -. ................................................................................... 104. -. ................................................................................................... 105 ...................................................................................................................... 108. .............................................................................................................................. 133.

(8) 1.

(9) -. Protein phosphatase 2A. 98% 400 protein kinase. [28, 34] PPP PPM Asp phosphatase. 40 [17]. protein phosphatase. Protein phosphatase 1 PP1. protein phosphatase 2A PP2A. PP5. type 2A protein phosphatase. PP4 PP6. PPP [130]. PP2A. 1%. PP1. 90%. [155]. PP2A [70] PP2A C. PP2Ac. A. PP2A. B. A ABC PR72/PR130. B B. B B55/PR55. B B56/PR61. PR93/PR110. 20 [125]. 2. A. B.

(10) B55 B56 C. B. PR72. A. 70. 72. G5pr. PR93 PP2A. Stm3. Stm. Stm4. A Protein phosphatase 2A PP2A PP2A. A B C. C. A. B 20. AC. 3.

(11) -. PP2A. hTERT telomerase catalytic subunit H-RAS SV40-ER [57] SV40-ER protein SV40-LT SV40-LT. small-t protein SV40-ST. p53 Rb. SV40-ST. PP2A hTERT H-RAS SV40-LT. SV40-ST [57] PP2A SV40-ST PP2A B56g3 B56g [117]. [27]. PR72. PP2A PP2A PP2A p53. Rb. cMyc [75] PP2A. PP2A [8, 9, 12, 16, 21, 22, 28, 43, 44,. 4.

(12) 69]. B. PP2A 1/3 HEAT. [113] P179R. PP2A A P179R. AC. B55. B56. [28, 146, 147] PP2A. CIP2A PME-1. SET. CIP2A B56g. cMyc. [141] CIP2A [135, 138] PP2A. PME-1. PP2Ac PP2A. [149] PME-1. ERK1/2 [110, 140] SET PP2A cMyc. ERK1/2 E2F1 [43, 69, 84, 88] SET [29, 43]. PP2A. PP2A. 5. Akt.

(13) SET. PME-1. CIP2A P cMyc. C. P Akt. B A. PP2A. P ERK1/2 P E2F1. Onco-protein. B PP2A PP2A. PP2A PP2A. SET PME-1 CIP2A. 6.

(14) PP2A PP2A PP2A. perphenazine PPZ PP2A. SET. PP2A. 7.

(15) perphenazine. 8.

(16) PP2A. PP2A PP2A. PP2A. PP2A perphenazine PPZ. [39, 54, 60, 107, 137] PPZ. T. PP2A A. PP2A [55]. T. SET. PP2A. [49]. PP2A. PPZ. T. PPZ. PP2A T. PPZ PPZ. 9. PPZ.

(17) - -. PPZ. T. T. T. PPZ. T. HUT78. CCRF-CEM. SKW3 A. T. Jurkat. TALL1. PPZ. T. PPZ. T. UL-1. Ema. PPZ. B. PPZ. T. PPZ PBMC. PPZ. PBMC Ema C. PPZ. PBMC PPZ. T. 10. Ema. UL-1 UL-1. PPZ.

(18) A. B 100. 120. 80. 80. 100. 60. 60. 40 20. HUT78 CCRF-CEM Jurkat TALL-1 SKW-3. 0 PPZ 0. 10. 20 20. 30 40. 0 PPZ. 50 µM. 60. *. 40 20. UL-1 Ema. 0. 0. 2.5 5.0 7.5 10 12.5 µM. PBMC Ema. UL-1. T. T. CCRF-CEM. *. 80. 40. PPZ A. C. 100. HUT78 Jurkat. 72. T. TALL1. SKW3. CCK8. PPZ. PPZ. 100%. N=6 B. T. Ema. 72. CCK8. 100%. N=6. C. PBMC. Ema. UL-1. PPZ PPZ. UL-1. PPZ. CCK8 100%. 72 PPZ. PPZ. N=4 *. 11. Ð< 0.05 PBMC.

(19) - -. PPZ. T. PP2A [49, 139]. PPZ. T HUT78. UL-1. PPZ. cleaved active. caspase 3. A-D. PPZ. cleaved caspase 3. Annexin V-FITC Apoptosis Kit Bio Vison PPZ. HUT78. FACS. UL-1 PPZ. Annexin V. PPZ E F. PPZ. HUT78. UL-1. 12.

(20) A. B. C. PPZ cleaved caspase 3 VCP. E. HUT78 0 12 24. hr. 100. 300. F. 200. 100. 100 0. 104 105 105.4 Annexin V. PPZ. 12. UL-1 0 12. 300 hr. 200 100 0 PPZ. 24 hr. 0. 12. hr. UL-1 PPZPPZ+. 300. 200. 103. 104 Annexin V. 105. T. A-D HUT78 A B 12. 0. 400 PPZPPZ+. 0 102.7. PPZ cleaved caspase 3 VCP. 200. 0 PPZ. *. 400. 300. HUT78. 400. D. *. 400. PPZ. 12 24. cleaved active caspase 3 A C. PPZ. UL-1 C D. PPZ. western blotting 100%. B D. N = 3 * Ð<. 0.05 PPZ E F HUT78 E Annexin V. UL-1 F. PPZ N = 2-3. 13. 12. FACS.

(21) - -. PPZ. T. HUT78. Akt. UL-1. PPZ. Akt Thr308. A-F. PPZ. Akt. Ser473. Akt Thr308. PDK1. [4] Akt Ser473. [123] PDK1. mTORC2. Ser241. [25]. mTORC2. Akt Ser473 mTORC2. UL-1. 15-30. PPZ. [51] HUT78. 15. PPZ. PDK1 NDRG1 PDK1. NDRG1. G-L PPZ. NDRG1. PP2A. PDK1. Akt. 14. mTORC2.

(22) B. C. 120 100. A PPZ p-Akt Thr308. 0. 100. 80. HUT78 15 30 60 120 min. 60 40. p-Akt Ser473. 120. *. *. 80. *. 60 40. 20. VCP. 0 PPZ 0 15 30 E. 60. 120min F. 120. D PPZ p-Akt Thr308. 0. p-Akt Ser473 VCP. 80 60. HUT78 PPZ p-PDK1 p-NDRG1 VCP. J. 120min. *. 80. *. 60 40. 20. 20 60. H 160 140 120 100 80 60 40 20 0 PPZ K 120. I. L. 160 140 120 100 80 60 40 20 0 PPZ 120 100 80. 60. 60. p-NDRG1. 40. 40. VCP. 20. 20. 0 PPZ. 0 PPZ. 15. **. 0 PPZ 0 15 30. 120min. 80. PPZ p-PDK1. 60. *. 120. 100. UL-1. 0 PPZ 0 15 30. 40. 0 PPZ 0 15 30 G. *. 100. 100 UL-1 15 30 60 120 min. **. 20. * 60. * 120min.

(23) PPZ A-F HUT78 15-120. T A-C. Akt PPZ. UL-1. Akt Thr308. blotting. PPZ. Ser473 A D. Akt Thr308 B E. D-F. 0 min. western. 100%. Akt Ser473 C F. N = 3 * Ð<. 0.05 0 min G-L HUT78 G-I PDK1. PPZ. UL-1 J-L. NDRG1 G J. NDRG1 I L. 0 min. PPZ. 15. western blotting. 100%. PDK1 H K. N=3 16.

(24) - -. PPZ. T. ERK1/2. Akt. ERK1/2 HUT78 ERK1/2. PP2A. UL-1. PPZ. PPZ. A-D. PPZ. ERK1/2 HUT78. UL-1. ERK1/2. MEK1/2. MEK1/2 [5] HUT78. Ser217. UL-1. MEK1/2. PPZ HUT78. Ser221. PPZ. 15. MEK1/2 ERK1/2. E-H PPZ. MEK1/2. B-Raf. C-Raf [93] I-N. HUT78. PPZ UL-1. B-Raf C-Raf B-Raf. PPZ. PP2A. MEK1/2 ERK1/2. 17.

(25) A. B. PPZ. 0. 15. HUT78 30 60 120 min. p-ERK1/2 VCP. C. D. PPZ p-ERK1/2. 0. 15. UL-1 30 60 120 min. VCP. E. 120 100 80 60 40 20 0 PPZ 120 100 80 60 40 20 0 PPZ. F. PPZ p-MEK1/2 VCP. *. PPZ p-B-Raf p-C-Raf VCP. UL-1 PPZ p-B-Raf p-C-Raf VCP. 0 1530. * 60. 120 min. * 120 min H UL-1. PPZ p-MEK1/2. K 120 100 80 60 40 20 0 PPZ. M. L. *. 120 100 80 60 40 20 0 PPZ. HUT78. 60. VCP. J. I. 0 1530. *. G 120 100 80 60 40 20 0 PPZ. HUT78. ** *. N 120 100 80 60 40 20 0 PPZ. *. 120 100 80 60 40 20 0 PPZ. 18. 120 100 80 60 40 20 0 PPZ. *.

(26) PPZ. T. A-D HUT78. A B. 15-120. ERK1/2 PPZ. UL-1. ERK1/2 A C. C D. PPZ. western blotting. 0 min 100%. ERK1/2. B D. N = 3 * Ð< 0.05 0 min E-H HUT78. E F. PPZ. MEK1/2 E G. 0 min. UL-1. G H. PPZ. 15. western blotting. 100%. MEK1/2. F H. N=. 3 * Ð< 0.05 0 min I-N HUT78. I-K. PPZ. B-Raf. C-Raf I L. C-Raf. UL-1. K N. L-N. PPZ. 15. western blotting. 0 min 100%. B-Raf. N = 3 * Ð < 0.05 0 min. 19. J M.

(27) - -. PPZ. PP2A. PPZ. Akt. Akt Ser473. ERK1/2 Thr202/Tyr204 A B. [47]. ERK1/2. PP2A. okadaic acid OA. PP2A. HUT78 PPZ. Akt. PP1. [64, 157]. PP1. tautomycin TAU [37] PP2B UL-1 TAU. PP2A. PP2B. ERK1/2. C D. OA. PP2A. ERK1/2. HUT78. PPZ. OA. PPZ ERK1/2. UL-1. Akt Ser473. ERK1/2. PPZ. PP2A. CsA. ERK1/2 [119, 159]. PP2B. PP1. cyclosporin A CsA [79] PPZ. PPZ PP2C. ERK1/2 ERK1/2. DUSP1/6. PP2C. BCI [97]. sanguinarine SA [1] DUSP1/6 HUT78. ERK1/2. UL-1 E F. SA. PPZ. 15-30. BCI. PPZ. ERK1/2 Na3VO4. PPZ. ERK1/2 G H. 20. Na3VO4. PPZ.

(28) ERK1/2. PPZ. ERK1/2. 21.

(29) A PPZ p-Akt Ser473. B. HUT78 Control 0 15 30. 0. OA 15 30 min. PPZ p-Akt Ser473. p-ERK1/2. p-ERK1/2. VCP. VCP. C. D. HUT78. Control PPZ 0 15 30 p-ERK1/2. TAU 0 15 30. CsA 0 15 30 min. PPZ p-ERK1/2. UL-1. PPZ p-ERK1/2. VCP. E. UL-1 Control OA 0 15 30 0 15 30 min. Control 0 1 2. TAU 1 2. 0. 0. CsA 1 2 hr. VCP HUT78 Control Sanguinarine 0 15 30 0 15 30 min. F. HUT78. PPZ p-ERK1/2. Control 0 15 30. BCI 15 30 min. 0. VCP. Long exposure. ERK1/2. HUT78. H. VCP. 140. #. *. 120 G PPZ p-ERK1/2. 100. HUT78 Na3VO4 Control 0 15 30 0 15 30 min. 80 60 40. VCP. 20 0 PPZ. 22. 0. 15 30 Control. 0. 15 30 min Na3VO4.

(30) PPZ. PP2A. Akt. A B HUT78. UL-1. PPZ HUT78. ERK1/2. ocadaic acid OA 100 nM. UL-1. 15-30. ERK1/2. Akt Ser473. western blotting. C D HUT78. UL-1. tautomycin TAU PPZ HUT78. ERK1/2 E HUT78. N=3 cyclosporin A CsA 10. UL-1. 15-30. western blotting. N=3. sanguinarine. 15-30. PPZ. ERK1/2. western blotting. N=. 2 F HUT78. BCI ERK1/2. G H HUT78 15-30. 1. PPZ western blotting. Na3VO4. 1. ERK1/2 G. Na3VO4. 15-30 N=2 PPZ. western blotting PPZ 0 min 100%. */# Ð<0.05 23. H. N=3.

(31) - -. Akt. T. PPZ. T. Akt. ERK1/2. Akt. Akt inhibitor VIII ERK1/2. FR180204 MEK1/2 HUT78. FR180204. UL-1. Akt inhibitor VIII. U0126. Akt. inhibitor VIII. T. FR180204. A-D. HUT78. U0126. UL-1 E F HUT78. U0126 UL-1. ERK1/2 H. U0126. U0126. Akt ERK1/2. CREB U0126. G. MEK1/2. ERK1/2 CREB HUT78 UL-1. Akt. HUT78. 24. Akt. UL-1. U0126. Akt.

(32) C. A. E. 100 80. 120. HUT78. 140. HUT78. 120. 120. 100. 100. 80. 80. 60. 0. 10. 30. 20. 20 M. B. 0 FR180204 0. 10. 140. UL-1. 30. UL-1. 100 80. 0 U0126. M. 300. 120. 250. 100. 200. 80. 60. 0 Akt inhibitor VI. 0. G. 10. 30. 0 FR180204 0 HUT78. 120 HUT78. U0126 p-Akt. 10. VCP. M. 0 U0126. U0126 p-Akt. *. *. 5. 10. M. UL-1. 80. p-ERK. 60. p-CREB. 40. 20. VCP. 20. U0126. UL-1. M. 100. 40. 0. 0. 120 UL-1. 100. 60. p-CREB. 30. H. 80. p-ERK. 10. 50. 20 M. 5. 100. 40. *. *. 20. *. 150. 60. 40. 0. F. D 120. *. 40. 40. *. 20 0 Akt inhibitor VI. 60. 60. *. 40. HUT78. 0. Ð. U0126. Ð. Akt A-F HUT78 A B. A C E. FR180204. UL-1 C D. B D F. Akt inhibitor VIII 0-30. U0126. E F. CCK8. 72 100%. N = 4 * Ð< 0.05 G H HUT78. G UL-1 ERK1/2. H. U0126 CREB. N = 1-3. 25. 24. Akt Ser473 western blotting.

(33) PPZ T [55]. PPZ. PP2A A. PP2A PP2A. T PPZ. PP2A. Akt. - HUT78. T. T. PPZ T. SKW3. MEK1/2. HUT78. MEK1/2 MEK1/2. HUT78. PPZ. UL-1 UL-1. HUT78. ERK1/2. Akt MCF7. MSTO-211H. 26. ERK1/2. Ema.

(34) MEK1/2. Akt. HUT78. [3, 101] MEK/ERK. MEK1/2. Akt. Akt UL-1. MEK1/2. Akt UL-1 MEK1/2. MEK1/2. ERK1/2. CREB. MEK/ERK. MEK1/2 Akt Akt. - -. PPZ. [41, 82, 152] Caspase 3 poly ADP-ribose polymerase PARP [46] cleaved active caspase 3. PPZ PPZ PP2A. SET. PP2A. FTY720. cleaved caspase 3. 27. OP449.

(35) [26, 49] PP2A. PP2A PP2A. [63, 90]. LB100. PP2A. FTY720. temozolomide. PP2A. SET. PP2A. [118] PPZ. T. Akt [111] PI3K/Akt [18, 50]. Akt. HUT78. UL-1 Akt idelalisib. pan-PI3K [150] PI3K/Akt. PPZ. Akt Thr308. 28. Ser473. copanlisib.

(36) PPZ. Akt. Akt Thr308. Ser473. mTORC2. PDK1. [4, 123] Akt. TSC1/2. [68]. Thr308. Akt Ser473. FOXO1 [68]. PPZ. PDK1. Akt Akt. mTORC2. PP2A. PPZ. PP2A. B56g. Akt. [7, 116, 143] PPZ. PP2A. Akt. Akt. Bcl-2 Bcl-2 Bax. Bcl-2. Bcl-xL. Bad [2]. Bcl-xL. Bax. Bad [38, 94, 145]. PPZ. Bcl-2. 29. Bcl-2. Bcl-2.

(37) - -. PPZ PPZ. D1 D2 [107]. D2 D3 D4. D1 D5 [11] T. [15, 158] D1 D5 ERK1/2. [102]. protein kinaseA PKA D2 D3 D4. ERK1/2. PKA. [14] PP2A. Ras. D2. Akt. PP2A. Akt. [112]. Raji Namalwa Daudi Jijoye dopamine. [15]. C-Raf B-Raf. MEK1/2 ERK1/2 PPZ. ERK1/2 PPZ. [107] D2. D2. PPZ PPZ. Akt. Akt. PPZ. - -. PPZ. ERK1/2. ERK1/2 MEK1/2. Bad. 30.

(38) FOS. CREB [8, 13, 87]. ERK1/2. HUT78. UL-1. PPZ. ERK1/2 MEK1/2. ERK1/2. PP1. PP2A PP2B PP2C DUSP1/6. [64, 76, 78, 157, 159] PPZ. ERK1/2. PPZ. ERK1/2 Na3VO4. PPZ. ERK1/2. ERK1/2. DUSP4 [78]. shRNA. PPZ. DUSP4 ERK1/2. - PPZ [59, 86]. PPZ. 31.

(39) PP2A small molecular activator of PP2A SMAP. [58, 73, 91, 113, 122, 134] SMAP. [122] SMAP. PPZ. Akt ERK1/2 PP2A. PPZ. T PP2A. Akt. SMAP SMAP T. 32.

(40) SET. 33.

(41) - -. PP2A. SET. SET I2PP2A TAF-1 [85]. ÍÛÌóÝßÒ ÍÛÌóÒËÐîïì SET. ERK1/2. Akt. [34, 100, 114, 115, 131]. [120]. 94% [148] PP2A. [120] [77]. [16] [43]. 34.

(42) - -. SET. SET. PP2A BCR-ABL. JAK2/PI3K/PKC. SET. SET. PP2A [100, 103]. SET. PP2A. Akt ERK1/2. cMyc [45]. p53 [131]. E2F1 [43] SET p70S6K [72]. SET SET. PP2A PP2A. PP2A. - -. SET. PP2A. Apolipoprotein E ApoE 178 277 [31] COG112 SET. OP449 COG449 PP2A. 35. ApoE. SET [83, 129].

(43) [45, 99]. FTY720 FTY720. SET. PP2A. [103] FTY720 [32, 98, 105, 115] T. OP449 [42, 49, 72]. FTY720. SET. PP2A. - -. SET T. SET. SET B. PP2A. [30, 33, 88, 126,. 142] SET. PP2A. Akt. Bid [80]. SET. 36.

(44) [53]. - SET. PP2A. SET SET. 37.

(45) - -. SET POS. HM-POS. SET. PP2A. SET. [100] SET. POS. PP2Ac. HM-POS SET. PP2Ac. A B POS. HM-POS. SET. SET. shRNA C D. shSET. SET SET SET. POS. HM-POS. E SET. POS F G. SET. POS. HM-POS. 38. HM-POS.

(46) A. B. C. 120 100 80 60 40 20 0. SET PP2Ac VCP. 120 100 80 60 40 20 0. D POS HM-POS shNT shSET shNT shSET. SET VCP. E. F. 1000 800. shNT shSET. 800. 600. ö. 400 200 0. 1000. POS. ö Day1 Day2 Day3 Day4. 120. shNT shSET. 600 shNT shSET HM-POS. 200 0. ö NT SET. POS. shSET. 120 100. 80. 80. 60 40 0. Day1 Day2 Day3 Day4. 39. 120 100 80 60 40 20 0. 100. ö. 20. shNT. HM-POS. POS. ö NT SET. G. POS. HM-POS. 400. 120 100 80 60 40 20 0. 60. HM-POS. ö. 40 20. shNTshSET. 0. shNTshSET.

(47) SET A B POS. HM-POS A B. SET PP2Ac. POS. SET. PP2Ac. 100. N=3. C D POS. HM-POS. shNT. shSET. SET. C. 100. D. E POS. HM-POS Day1-4. Day1. F G POS. N = 3 * Ð< 0.05 shNT shNT. shSET. CCK8. 100. shNT N = 6 * Ð < 0.05. HM-POS. shNT. shNT. shSET F. 100 N = 6 * Ð< 0.05 shNT. 40. shNT. shNT G.

(48) - -. SET. SET. PP2A. ERK1/2. mTORC1. Akt. [45, 72, 131] SET POS. SET HM-POS. ERK1/2 SET. ERK1/2. p70S6K SET ERK1/2. mTORC1 A-H. POS. ERK1/2. mTORC1. 41. HM-POS.

(49) A. B. POS shNT shSET. p-ERK1/2 ERK1/2 VCP C. POS shNT shSET. p-MEK1/2 MEK1/2 VCP E. POS shNT shSET. p-p70S6K p70S6K VCP G. 120 100 80 60 40 20 0. ö NT SET. 120 100 80 60 40 20 0. p-ERK1/2 ERK1/2 VCP NT SET D. 140 120 100 80 60 40 20 0. NT SET. 140 120 100 80 60 40 20 0. shNT shSET. p-S473Akt Akt. MEK1/2 VCP. NT SET F. 300 250 200 150 100 50 0. NT SET. 180 160 140 120 100 80 60 40 20 0. HM-POS shNT shSET. p-p70S6K p70S6K VCP. NT SET H. 140 120 100 80 60 40 20 0 140 120 100 80 60 40 20 0. VCP. HM-POS shNT shSET. p-MEK1/2. POS. p-T308Akt. HM-POS shNT shSET. NT SET. 120 100 80 60 40 20 0. 140 120 100 80 60 40 20 0. 120 100 80 60 40 20 0. ö NT SET. NT SET. ö NT SET. 120 100 80 60 40 20 0. 140 120 100 80 60 40 20 0. 120 100 80 60 40 20 0. ö NT SET. ö NT SET. NT SET. HM-POS shNT shSET. p-T308Akt p-S473Akt Akt. NT SET. 120 100 80 60 40 20 0. 140 120 100 80 60 40 20 0 140 120 100 80 60 40 20 0. VCP. NT SET. NT SET. 120 100 80 60 40 20 0. NT SET. NT SET. SET POS. A C E G. HM-POS. B D F H. ERK1/2 (A B) MEK1/2 (C D). shNT. 100. * Ð< 0.05 shNT. 42. shNT. shSET. p70S6K (E F) Akt (G H). N=3.

(50) - -. SET. SET. PP2A. E2F1. cMyc. [43, 45, 127] SET. SET. POS HM-POS. E2F1. cMyc. A-F. POS. HM-POS. SET. SET. ERK1/2. ERK1/2. mTORC. 43. E2F1.

(51) POS shNT shSET. HM-POS shNT shSET. POS shNT shSET. HM-POS shNT shSET. POS shNT shSET. HM-POS shNT shSET. p-E2F1 E2F1 VCP. p-cMyc cMyc VCP. SET POS. A C E. HM-POS. E2F1 A B. B D F. C D. shNT. shSET. cMyc E F shNT. 100. N = 3 * Ð < 0.05 shNT. 44.

(52) - -. ERK1/2. POS. ERK1/2 POS. ERK1/2 POS. HM-POS. ERK1/2. FR180204 POS. HM-POS POS. ERK1/2. A-C HM-POS POS. ERK1/2. 45.

(53) A. B. C POS. ö. 120. ö. 120. 100. 100. 80. 80 FR180204. 60. 100 µM HM-POS. 60. 40. 40. 20. 20. 0 FR180204. 100 µM POS. 0 FR180204. 100 µM HM-POS. 100 µM. FR180204. 100 µM POS. 100 µM HM-POS. ERK1/2 A POS. HM-POS. FR180204. CCK8 100% B C POS. 72. FR180204 N = 6 * Ð< 0.05 FR180204 HM-POS. FR180204 B. 100%. 10 FR180204 C N = 6 * Ð<. 0.05 FR180204. 46.

(54) - -. SET SET SET. POS. HM-POS. FTY720. FTY720. FTY720. A cisplatin POS. HM-POS. FTY720 cisplatin FTY720. B. 47. cisplatin.

(55) A. B POS. POS 120. 120. 100. 100. 80. 80. 60. 60. 40. 40. 20. 20. 0 FTY720 0. 1. 3. 5. 0 Cisplatin FTY720. 10 µM. HM-POS. 120. 120. 100. 100. 80. 80. 60. 60. 40. 40. 20. 20. 0 FTY720 0. 1. 3. 5. 0 Cisplatin FTY720. 10 µM. ö #. 1 µM. 1 µM 5 µM. 5 µM HM-POS. ö. #. 1 µM 5 µM. 1 µM 5 µM. SET A POS. HM-POS. CCK8. 72. FTY720. 100% B POS. FTY720. N = 6 * Ð< 0.05 FTY720 HM-POS. FTY720. cisplatin. CCK8 100%. N = 6 */# Ð< 0.05. 48. 72.

(56) - -. SET. POS. HM-POS. SET SET. PP2A. [42, 49, 66]. POS. HM-POS. SET. PP2Ac. SET g SET. PP2A. 94%. [148]. [16]. SET [43] PP2A. [120] [77]. SET shRNA. SET SET SET. HM-POS. POS POS. [10]. 49.

(57) SET. POS. HM-POS. HM-POS SET. SET. [96]. HM-POS. SET. SET SET SET. PP2A. SET. - -. SET MEK/ERK. PI3K/Akt. [21] KRAS. EGFR [52]. HM-POS. ERK1/2. E2F1. mTOR ERK1/2. POS. PP2A [43, 72, 76] PP2A. SET ERK1/2. p70S6K E2F1 SET. 50. PP2A.

(58) SET. HM-POS. cMyc. 11. SET. E2F1. cMyc [43]. POS. E2F1. cMyc SET ERK1/2 SET. - -. SET. FTY720. cisplatin SET 11. SET. [43]. SET. SET Cisplatin DNA. 51. DNA.

(59) [35] SET FTY720. SET. cisplatin. doxorubicin. SET [72]. -. 27. [128] SET ERK1/2. p70S6K E2F1 SET. FTY720. cisplatin SET. SET. 52.

(60) SET. PP2A. 53.

(61) - -. PP2A/B56. PP2A. B. 20 B. 18. B56. HEAT. [147] B56. B56g3. B56g PP2A [27, 117, 141]. PP2A/B56. cMyc. Akt. p53 [75]. SMAP B56. A [122]. - -. SET. SET. PP2A A. PP2A/B56. PP2A. PP2Ac. PP2A. PP2A. [104]. B SET. SET. PP2A. PR72. 54.

(62) [43] p70S6K. SET. ERK1/2. E2F1 E2F1. ERK1/2. PP2A/B56g1. [43] p70S6K. PP2A/B56g. PP2A/B56g [19, 56, 76]. SET. PP2A. PP2A/B56. SET. PP2A. SET. SET. PP2A. - PP2A. SET. PP2A. SET. PP2A. SET SET PP2A SET. PP2A. 55.

(63) - -. SETa. PP2A/B56 SET. PP2A. B. FLAG. 293T. SET. B. A B. B56. SETa. B55. PR72. B56a B56b B56g2 B56g3 B56d B56e. PP2A SETa. B56 SmBiT. SET. B. 293T C. SmBiT 6. SETa. B56a. B56g3 B56g2 SETa GST-SETa. B56a His-B56a D. SETa. pull B56a. 56.

(64) A. C °<0.001. FLAG SET. 180 160 140 120 100 80 60 40 20 0 SmBiT. 37. PP2Ac FLAG. 75 50. SET. 37 37. PP2Ac. 75 50 100. FLAG VCP. B. ns (°=0.482). 37. °=0.004. 75 GSTPull Down. D. 300. Input. GST. 250 200. His. 150 100. 50 75. 50 GST. 0 FLAG. 25. 57.

(65) SETa. PP2A/B56. A B 293T. FLAG. B. FLAG M2. B. SET A. B56g3. SET. 100. C 293T SETa. FLAG B. SmBiT. B. 24 100. N=4 D. GST. GST-SETa. His-B56a. GST-SETa. His-B56a. N=4. 58. N = 3 Empty.

(66) - -. SETa. - - -. B56a. SETa. B56g3 B56a. B56g3. SETa B56 B56g3. B56a. SETa. B56. HEAT. A B C. B56a B56g3. N. SETa WT C-H. B56a. B56g3. N. C. SETa. N. PP2Ac. C B56a. PP2Ac. C. SETa PP2Ac. PP2Ac B56g3. SETa. C. SETa B56a. 59. B56g3. PP2Ac SETa.

(67) A FLAG-B56 WT. 59 77 91 110 119 131 408423 434 455 486 111. 1- 408 1- 423 1- 455. 486. 59. 408. 59. 423. 59. 455. B FLAG-B56 3 41 52 68 80 92 102 385396 409430 524 WT 41 524 92 1- 383 1- 408 1- 441. 41. 524 383. 41. 408. 41. 441. C. F. FLAG-B56 SET PP2Ac. 37 37 75. FLAG 50 SET. 37. PP2Ac. 37 75. FLAG VCP. 50 100 75. FLAG-B56 3. 60.

(68) SETa A B. B56a B56a. B56g3. B56g3. SETa. PP2Ac. WT C-H 293T F-H. FLAG. B56a. FLAG M2. B56g3. B56a B56g3. SET FLAG. C-E. C F WT. SETa D G. N=3. 61. PP2Ac E H. 100.

(69) - - -. B56a. B56g3. SET. SETa. CD. ED A. B56a. B56g3 SETa. AD. B AD. B56g3 SETa. SETa. B56. 62. B56a. SETa.

(70) A GFP-SET. 225 290 31 78 79 CD ED AD 31 78 CD 79. ED. 225 226. 290 AD. 31 78 79 225 CD ED 79. B. ED. C. 293T GFPc-SET. 226. 290 AD. 293T GFPc-SET. 100. GFP. 100. GFP 25. 25. 37. PP2Ac. 75. FLAG. 37. PP2Ac. 75. FLAG. 100. 100. GFP. GFP 25. A SETa. B56g3. CD. AD CE ED. B C 293T. CD. AD. EA. FLAG. B56a B. SETa WT CD ED AD CE EA B56a. 75. SETa B56a. ED. 100. VCP. 75. B56g3. 75. FLAG. 100. VCP. 37. PP2Ac. 75. FLAG. B56a. 25. 37. PP2Ac. B56g3 N=3 63. WT B56g3 C FLAG M2. SETa. ED. GFP.

(71) - -. SETa. B56a. B56g3. B56g3. SETa. SET SET. [95] SET. CD. B56g3 SETa. SETa DD. [95]. SETa DD. A. SETa DD B. SETa WT. SETa DD C. SETa DD. B56g3. SETa. SETa DD. B56g3 D E. SETa B56a. B56g3. SETa. SETa. B56a. SETa B. B56a F. SETa DD. B56a. SETa DD B55. PR72. SETa DD G A549. B56g2. B56a. SETa. B56a. B56a SETa WT. SETa DD H. 64. SETa.

(72) B56a. B56g3. PP2A/B56. 65.

(73) D. B. A SET DD. CD. ED. EmpWT WTWT EmpWT DD. AD. 78. 100. GFP. °<0.001 °<0.001. 25 50. FLAG. 100. 100. 80. GFP E. GFP. 75. 120 °<0.001. 80. 40 FLAG WT WT Empty WT. WT DD. DD WT. DD DD. 60. 25 50. 20. SmBiT-SET. 100. 100. 60. 0. 75. FLAG. °<0.001. 120. C. FLAG Empty GFP-SET WT DD WT DD 100 GFP 75 PP2Ac. VCP. 40. 100. 20. 75. 0 DD WT SmBiT Empty Empty. DD. WT. G. F FLAG Empty PR72 GFP-SET WT DD WT DD WT DD WT DD WT DD WT DD 100 GFP 75 PP2Ac. 100. °<0.001. 80 60. FLAG GFP. H. A549 Emp Emp. °<0.001. ns (° °=0.014. 120. °=0.015. 100 80 60 40 20 0 FLAG GFP. 120. Emp Emp. WT. 66. 75. 40. 50 100. 20. 75. 0 WT DD SmBiT Empty Empty. WT. DD.

(74) SETa. 2. B56a. B56g3. A SETa DD B 293T. FLAG. SETa DD. SETa WT. GFP. FLAG M2. SETa WT Empty. SETa WT SETa WT. SETa DD N=3. C 293T. DD SmBiT-SETa WT. DD. 24 100 N=3 D 293T. FLAG. SETa DD SETa WT Empty E 293T. B56g3. GFP. FLAG M2. SETa WT B. SETa DD N=3 DD. SmBiT-B56g3. 100 N=4 67. 24.

(75) F 293T PR72. FLAG. B. GFP. B55a B55d B56a B56g2. SETa WT. SETa DD. B. M2. SETa WT Empty. FLAG SETa DD N. =3 G 293T. DD. SmBiT-B56a. 100 N=4 H A549. FLAG-B56a. GFP-SETa WT. DD. 14 Empty. 100. N=4. 68. 24.

(76) - -. SETa. SETa SETa DD. B56a. B56g3. CD [95]. SETa SmBiT-SETa. A549-nSETa/cSETa-A2. A549. C6. 15. A. A549-nSETa/cSETa-C6 KN-93. MG132. I3 M. KN-93 52. 29. 28. KN-93. MG132. B. KN-93 C-E. PPI SmBiT-. 22 LSm-BiT. LSm-BiT. A549-LSm-G1. 69. PPI A549 KN-93.

(77) F KN-93. LSm-BiT 8. KN-93 9. A549-nSETa/cSETa-C6. SETa LSm-BiT. 25. A549-nSETa/cSETa-C6. 32 A549-nSETa/cSETa-C6. 70.

(78) 71.

(79) SETa A A549. SmBiT-SETa. A549-nSETa/cSETa-A2. C6 DMSO. 100. B A549-nSETa/cSETa-C6. KN-93. MG132. I3 M. DMSO. 100. N=4. C-E A549-nSETa/cSETa-C6. C. D KN-93 0.5-10. E DMSO. 100. N = 4-6. D * Ð <= 0.001. E * Ð<= 0.005 F A549. LSm-BiT. A549-LSm-G1. 5. KN-93 DMSO. 100. 4 72. N=.

(80) - -. KN-93. SETa/B56a SETa/B56g3. SETa. SETa-B56a A549. SETa-B56g3. SmBiT-B56a. A549-nSETa/nB56a. A549-nSETa/nB56g3. KN-93 KN-93 A B. SETa-B56a. SETa-B56g3. SETa. B56a. 73. B56g3. B56g3.

(81) A 120 100. 120. °<0.001 °<0.001. 100. 80. 80. 60. 60. 40. 40. 20. 20. 0. 0. KN-93 A B A549. °=0.002 °<0.001. SETa-B56a SETa-B56g3 SmBiT-B56a. A A549-nSETa/nB56a. SmBiT-B56g3 B:A549-nSETa/nB56g3. KN-93. DMSO. 100. N=4 74.

(82) - -. KN-93. SETa. SETa DGAT VCP [61, 124, 132] KN-93. [108, 144]. SETa A549-nSETa/cSETa-C6. KN-93. KN-92. W-7. KN-62 A-D. KN-92. SETa. KN-93 W-7 KN-62. SETa KN-93. Fyn Hck Lck MLCK. TrkA Lck MLCK. [108] TrkA. Fyn Hck SETa. CHR6494 CHR6494 KN-93 KN-93. 75. SETa. SETa SETa.

(83) KN-93. SETa. A-E A549-nSETa/cSETa-C6. A KN-92. B W-7. C KN-62. D. E CHR6494 DMSO 0.023. 100. N=4. C * Ð= 0.002 76. B * Ð=.

(84) - -. KN-93. KN-93. SETa. SETa A549-nSETa/cSETa-C6 KN-93. A-C. KN-93. SETa KN-92. SETa. KN-93 KN-93. LSm-BiT D. KN-93. SETa SETa LSm-BiT. E F. SETa SETa. 77.

(85) 78.

(86) KN-93. SETa. A A549-nSETa/cSETa-C6 KN-93 DMSO. 100. N = 4 * Ð< 0.01. B A549-nSETa/cSETa-C6 KN-93. -. 100. N = 4 * Ð< 0.01. C A549-nSETa/cSETa-C6 KN-92 DMSO. 100. N=4. D A549-LSm-G1 KN-93. DMSO. 100. N = 4 * Ð< 0.05. E A549-nSETa/cSETa-C6. DMSO. 100. N=4. F A549-LSm-G1. DMSO. 100. N=4. 79.

(87) - -. SETa. PP2A. SET [85]. ÍÛÌóÝßÒ ÍÛÌóÒËÐîïì PP2Ac. B. PP2A I2PP2A. PP2A SET. [6, 84] PP2Ac. 95~112 133~146. SETb. 165~182. SETb. PP2Ac. [133] SET. SET. PP2Ac. PP2A SET. È»²±°« ´¿»ª PR48. 1~18 151~172. B55d B56g. [62]. B56a. B55a. SET. PR72. [43] SETa. B56a. B56. B56g3 B56g2. SET. SETa. B56. SET [118, 151]. B56a. 80. [48] B56g2.

(88) [67] B56g3. [92]. B56a B56g3. SETa. B56 SETa. B56. PP2Ac. B56g3 PP2Ac B56a. - -. C. PP2Ac. SETa. B56g3. SET. SETa. PP2A. SET. SET. SETb importin [151]. PKC. [103, 154]. CD SETa. 37. SETb. 11. N. 1. 1 24 SETa. v6.5.7 2019. CK2. N. 25. SETa. SETa. 81.

(89) SET. PP2A. [103] FTY720. PP2A SETb. CD. SETb SET. Gln29 Lys77. PP2Ac. [36]. PP2A FTY720. SETa. N FTY720. SET. SETa. SETb. SETa N SETb. FTY720 FTY720. PLA. SETb. PLA. in vivo. FTY720. SET in vitro. A549. SET. PP2Ac. B56a SETa WT. 82.

(90) SETa DD SETa. PP2A/B56a. A549 B56g3. B56g [117]. SETa. B56g3. SETa. KN-93. A549. KN-93 SETa SETa. KN-93. - -. SETa. B56. B56a B56g3 B56a. SETa AD SETa. SETa. AD. EA. CD. B56a. SETa DD. N. B56a. B56g3. SETa. B56a. SETa. SETa AD. B56a. AD B56g3. B56a SETa CD. 83. CE.

(91) SETa. AD. B56. SETa. SETa SETa. B56. SETa. - -. KN-93. SETa. SETa KN-93. SETa [108, 144]. W-7 KN-62. SETa. in vitro. KN-93 SETa. KN-93. SETa. SETa LSm-BiT. 60%. 25%. SETa LSm-BiT. SETa SETa. 84. 50%.

(92) MEK/ERK SETa PP2A. [71]. SETa SETa. PP2A/B56 B56a B56g3. KN-93. SETa SETa SETa. SETa-B56a SETa-B56g3. PP2A/B56 PP2A/B56. SETa. 85.

(93) 86.

(94) -. PP2A PP2A. PP2A. PID. PAD. SET-PP2A. FTY720. SMAP. PAD. PP2A. SMAP. OP449. PP2A A. PP2A. PID. PPZ. LB100. HEAT /. SMAP. SMAP. P179R [58] SMAP. Akt [134]. SMAP. PP2A A. PP2A B56 [122] PPZ. SMAP. PP2A SMAP. ERK1/2 ERK1/2. PP2A SMAP. SETa. PP2A/B56. PP2A B56a. B56g. CIP2A [141] SMAP. CIP2A. 87. PP2A. SET.

(95) FTY720. PP2A. SETb. [36] FTY720. S1P. [20] FTY720 SPHK2. S1P. [1]. S1P FTY720 [24]. SET. FTY720. [103] SPHK2. FTY720. [105]. FTY720. OSU-2S. S1P. CM-1231 PP2A [139]. FTY720 SETa. PP2A/B56. KN-93. SETa SETa. SETa SETa. SETa KN-93. SETa. 88.

(96) PID. LB100. PP2A. LB100. PP2A [81] PP2A. PP2A [136]. LB100. PP2A. PP2A B. PP2A B55a. cMyc Thr58. 66cl4. [153]. B55a. Thr41 [81, 156] GSK3b. PR130 [40]. LB100. PP2A PP2A LB100 PP2A/B55a [81]. LB100. PP2A. PP2A PP2A. PP2A SMAP. SET. PAD. 89.

(97) -. SET SET. [16, 43,. 49, 65, 66, 72]. SET BCR-ABL. JAK2. SET. [100] RUNX1 GATA2. Myc SP1. SET. SET mRNA [109]. Myc. RUNX1. [89, 106] Wnt. ÕïçóÉ²¬ï. COX2-PGE2. ÕïçóÝî³Û. Ù¿² mRNA. [43]. ÕïçóÉ²¬ï SET. ÕïçóÝî³Û. mRNA. SET mRNA. Ù¿². SET Ù¿² SET mRNA. SET. SET. SET SET. 90. SET.

(98) SET [43]. SET E3 /. proteolysis targeting chimeric molecules PROTACs [121] KN-93. SET. PROTACs. SET. A549. SETb. CK2. Ser9. SET. -. PP2A SET. [43] SET. 91.

(99) SET. OP449 [43] SET. aurora kinase [74]. OP449. SET SET. 92.

(100) 93.

(101) T. HUT78. SKW3. Jurkat. T. TALL1. Ema. T. UL-1. PBMC. HM-POS. POS. AA Thermo Scientific FBS SIGMA. A549. CCRF-CEM. 293T. RPMI1640. Takara Bio. 1%AA 10% FBS. Eagle s medium DMEM SIGMA MCO-5AC SANYO. 10% fetal. 37°C 5% CO2 SCA-80D ASTEC HBS 25 mM HEPES 150. mM NaCl pH 7.4 acid EDTA. -. 0.05% trypsin 0.53 mM ethylenediamine tetraacetic. HBS. DNA. - N. 3×FLAG. B56b. PR72. cDNA. N. 3×FLAG PCR InFusion HD Cloning Kit Takara Bio. 94.

(102) C. GFP. DD. DD Cloning Kit. PCR. pLVGFPc. N. N. InFusion HD GST. 8×His. pLVSIN PCR. pGEX4T1. Cloning Kit SmBiT. N. InFusion HD. pHEX1. LgBiT. C. WT PCR pBiT1.1-C[TK/LgBiT]. InFusion HD Cloning Kit. pBiT2.1-C[TK/SmBiT]. pBiT1.1-N[TK/LgBiT]. pBiT2.1-N[TK/SmBiT] shRNA pLVmC. ClaI/MluI. T4 DNA ligase Takara Bio. shRNA Non-target shRNA shNT. 5 -CAACAAGATGAGAGCACCA-3. SET target shRNA shSET. 5 -GGATGAAGGTGAAGAAGAT-3. - XL10GOLD Stratagene 0.5 µL. HST08 Takara Bio 10 µL. 30 42°C. Major Science. 45. 50 µL. 95. LB.

(103) Broth. 50 µL. super optimal broth with catabolite repression SOC Medium. 37°C. BioShaker BR-22FP Taitec. 100 µg/mL ampicillin. LB agar. 37°C LB Broth 100 µg/mL ampicillin. 200 rpm. 37°C 4,000 rpm. 2420 KUBOTA. 10. Plasmid Mini. Kit I Omega. - 12. 2×105 cell/well. 293T. Opti-MEM. 1.0 µg. 2.5 µL. 30. 200 µL. Polyethylenimine MAX PEI Polysciences 1 mL. 10% FBS 1%AA. DMEM. 24. - 293T. 4×105 cell/well packaging plasmid psPAX2. plasmid pMD2.G 30. 2.5 µL 1.5 mL. PEI. VSVG coat protein Opti-MEM. 10 FBS 1 AA. 1.5 mL. 40. 96. DMEM.

(104) 0.22 µm 12. shRNA. BL21 DE3 Codon Plus RP Agilent Technologies 50 uL 30 LB Broth 100 µg/mL ampicillin 35 µg/mL chloramphenicol 37°C 250 rpm. 200 mL. Broth 100 µg/mL ampicillin 35 µg/mL chloramphenicol. LB. 1L. 37°C 200 rpm. IPTG isopropyl 0.2 mM. 37°C 200 rpm. 3,000 rpm 15. 4°C. recombinant buffer A 20 mM Tris HCl pH 7.5 150 mM NaCl recombinant buffer B. 20 mM Tris HCl pH 7.5 150 mM NaCl 0.1. mM EGTA 5 mM benzamidine 1 mM PMSF 1 mM DTT 0.1% Tween 20 20 mM Tris HCl pH 7.5 150 mM NaCl 0.1 mM EGTA 5 mM benzamidine 1 mM PMSF 20 mM imidazole 0.1%Tween 20 10 ml. lysozyme 4°C. 12,000 rpm. 10. glutathione sepharose. KUBOTA GE Healthcare. 97. HisPur Ni-NTA Thermo.

(105) Scientific. 4°C 30. imidazole. GST/His. PD-10 GE Healthcare. recombinant storage buffer 20 mM Tris HCl. pH 7.5 150 mM NaCl 0.01% Tween 20 coomassie brilliant blue CBB CBB. CBB. 50% methanol 10% acetic acid. -. CBB. 0.1% coomassie brilliant blue 7.5% acetic acid 5% methanol. Western blotting. - 1,500 rpm Lysate buffer 50 mM Tris-HCl pH 8.0 5 mM EDTA 5 mM EGTA 1 mM Na3VO4 20 mM sodium pyrophosphate 1% Triton X-100 1 tablet/50 mL Roche Complete protease inhibitor. HBS. lysate buffer °C 15,000 rpm. - -. 15. Western blotting DC protein assay kit Bio-Rad. Lowry. 10-40 µg. 98. SDS.

(106) 8-15%. Bio-Rad Wako. 0.5%. 3%. PVDF. Bio-Rad. PVDF TBS-T 25 mM Tris-HCl pH 7.4 150 mM NaCl. 0.05% Tween 20. 30-60. °C. 1 Reagent PerkinElmer. ECL Pro western blotting Detection LAS3000mini FUJIFILM. ImageJ National Institutes of Health p97/VCP. - -. pull down assay FLAG M2 HBS. lysate buffer. 40 mM HEPES pH 7.4. 2 mM EDTA. CHAPS 10 mM disodium. glycerophosphatate 0.3% CHAPS 10 mM sodium pyrophosphate 2 tablet/50 mL PhosSTOP 1 tablet/50 mL Roche Complete protease inhibitor °C 15,000 rpm. 15. CHAPS wash buffer 40 mM HEPESpH 7.4 2 mM EDTA 10 mM disodium glycerophosphatate 0.3% CHAPS 10 FLAG M2 Affinity Gel SIGMA. mM sodium pyrophosphate °C. 30. SDS. 99.

(107) sample buffer 50 mM Tris-HCl pH 6.8 0.4% SDS FLAG peptide SIGMA. Glycerol. - -. 2%. western blotting. GST pull down assay 100 pmol. GST-tag 100 pmol. glutathione sepharose. 10 pmol. recombinant buffer A. °C 30. SDS sample buffer. western. blotting. Cell Counting Kit-8 Dojindo 4.0×104. 96. PPZ. 450 nm. 72 24. 1.0×104 FTY720. cisplatin. 72. 450 nm. 1.0×102 RPMI1640. 2 mL 10% FBS 1% AA 10. 10. 100. DMEM.

(108) -. NanoBiT. - 293T 24. NanoBiT 1.0×104. 96. LgBiT100 ng. Perkin Elmer 6005181. SmBiTPEI. Opti-MEM 24. Substrate Nano-Glo LCSDilution Buffer 25 µL. 10. 37°C. Nano-Glo Live Cell. 5 95. % CO2. 37°C. ARVO X PerkinElmer. GloMAX Promega. 10. NanoBiT. SmBiT-. LgBiT-. 30. 101. N. C.

(109) B55. B56. 120. 120. 100. 100. 80. 80. 60. 60. 40. 40. 20. 20. 0 LgBiT SmBiT. 0 LgBiT SmBiT. B56. B56. 120. 120. 100. 100. 80. 80. 60. 60. 40. 40. 20. 20. 0 LgBiT SmBiT c56. c56 n56. c56. c56. n56. n56. n56. 0 LgBiT SmBiT c56. c56 n56. c56. c56. n56. n56. n56. NanoBiT. 293T. LgBiT-. SmBiTSmBiT-. empty vector. 24. empty vector 100. Duplicate. 102.

(110) - -. NanoBiT. A549. PCR pLVnB2puro. pLVnB2neo. - A549. NanoBiT. - -. NanoBiT. A549. A549. 96 2.0×104. A549. 24. 20 Promega. CellTox Lysis Solution. 10 µL. Nano-Glo Live Cell. Substrate CellTox Green Nano-Glo LCS Dilution Buffer 25 µL. 10. 37°C. 37°C. 5 1 94. % CO2 ARVO X PerkinElmer. 10 618 nm. 100%. 103.

(111) -. -. (. )/. I. - -. III. In vitro NanoBiT A549 CHAPS IPBuffer. 15,000 rpm. 15. in vitro NanoBiT. in vitro. 96. NanoBiT. 1.0×105. Thermo Scientific 611F96WT. 4°C. 0.625 11.74. Nano-Glo Live Cell Substrate Nano-Glo LCS Dilution Buffer 12 µL GloMAX Promega. 10 10. 10. 100%. HUT78. PPZ. 24. UL-1. 104. PPZ. 12.

(112) 1,500 rpm Annexin V-FITC Apoptosis Kit Bio Vision. Annexin V. Flow cytometry FACS BD Accuri BD Biosciences Annexin V. ± t-test. Student s One way ANOVA. Significant Difference LSD test. P. 105. Fisher s Least 5%.

(113) p-Akt Ser473. Cell Signaling. 4060. p-Akt Thr307. Cell Signaling. 13038. Akt. Cell Signaling. 2920. p-B-Raf Ser445. Cell Signaling. 5284. p-C-Raf Ser338. Cell Signaling. 9427. cleaved caspase 3 Asp175. Cell Signaling. 9664. caspase 3. Gene Tex. GTX11-543. p-cMyc Ser62. Bio Academia. 71-161. cMyc. Santa Cruz. sc-764. p-CREB Ser133. Cell Signaling. 9198. p-E2F1 Ser364. MBL. D246-3. E2F1. Santa Cruz. sc-193. p-ERK1/2 Thr202/Tyr204. Cell Signaling. 9101. ERK1/2. Cell Signaling. 9107. FLAG. Sigma. F7425. GFP. Santa Cruz. sc-9996. p-GSK3b Ser9. Cell Signaling. D85E12. GSK3b. Santa Cruz. sc-9166. GST. ABGent. 9AT106. His. Gene Tex. GTX115044. p-MEK1/2 Ser217/Ser221. Cell Signaling. 9154. MEK1/2. Cell Signaling. 4694. p-NDRG1 Thr346. Cell Signaling. 5482. p-PDK1 Ser241. Cell Signaling. 3438. PDK1. Cell Signaling. 3062. p-p70S6K Thr389. Cell Signaling. 9234. 106.

(114) p70S6K. Santa Cruz. sc-230. PP2Ac. Millipore. 07-324. SET. Santa Cruz. sc-5655. SET. Santa Cruz. sc-133138. p97/VCP. GeneTex. GTX113030. / Perphenazine. SIGMA. Tautomycin. Wako. CycrosporineA. Wako. Sanguinarine. EXTRASYNTHESE. BCI. Merck Millipore. FR180204. SIGMA. FTY720. Cayman Chemical. Cisplatin. Wako. Xanthohumol. Cayman Chemical. KN-93. Wako. W-7. Cayman Chemical. KN-62. Cayman Chemical. CHR6494. Tocris Bioscience. KN-92. R&D SYSTEMS. 107.

(115) apoptosis via phosphorylation of p38 in HL60 cells. Þ± ½ ò Þ±¬»½¸²±´ò Þ±½¸»³ò éì: 548 552.. family. Ì®»²¼ Þ ±½¸»³ò Í½ ò îê: 61 66.. Þ ±½¸»³ò Í±½ò Ì®¿² ò ìð: 139 146. 4. Alessi, D. R., James, S. R., Downes, C. P., Holmes, A. B., Gaffney, P. R., Reese, C. B. and Cohen, P. 1997. Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activates protein kinase Balpha. Ý«®®ò Þ±´ò ÝÞ. é: 261 269.. Ashworth, A., Marshall, C. J. and Cowley, S. 1994. Identification of the sites in ÛÓÞÑ Öò ïí: 1610 1619. 6.. Amick, G. D., Reddy, S. A. and Damuni, Z. 1992. Protein phosphatase 2A is a specific protamine-kinase-inactivating phosphatase. Þ ±½¸»³ò Öò îèé: 1019 1022.. 7.. Arroyo, J. D. and Hahn, W. C. 2005. Involvement of PP2A in viral and cellular transformation. Ñ²½±¹»²». îì: 7746 7755.. ERK1/2 pathway. Ý»´´ Ü»¿¬¸ Ü ºº»®ò ïê: 368 377. 9.. Baril, C., Sahmi, M., Ashton-Beaucage, D., Stronach, B. and Therrien, M. 2009. 108.

(116) signaling in Drosophila. Ù»²»¬ ½ . ïèï: 567 579.. and characterization of the growth and pulmonary metastasis of a highly lung metastasizing cell line from canine osteosarcoma in nude mice. Öò Ê»¬ò Ó»¼ò Í½ ò êï: 361 367. 11. Beaulieu, J.-M., Espinoza, S. and Gainetdinov, R. R. 2015. Dopamine receptors IUPHAR Review 13. Þ®ò Öò Ð¸¿®³¿½±´ò ïéî: 1 23. 12. Bhaskar, P. T. and Hay, N. 2007. The two TORCs and Akt. Ü»ªò Ý»´´. ïî: 487 502. 13. Bhattacharjee, A., Mulya, A., Pal, S., Roy, B., Feldman, G. M. and Cathcart, M. K. Monocyte 15-Lipoxygenase Gene Expression Requires ERK1/2 MAPK Activity. http://www.jimmunol.org. 14. Bozzi, Y. and Borrelli, E. 2013. The role of dopamine signaling in epileptogenesis. Ú®±²¬ò Ý»´´ò Ò»«®± ½ ò é: 157. 15. Braesch-Andersen, S., Paulie, S. and Stamenkovic, I. 1992. Dopamine-induced Í½¿²¼ò Öò ×³³«²±´ò íê: 547 553. 16. Brander, D. M., Friedman, D. R., Volkheimer, A. D., Christensen, D. J., Rassenti, L. Z., Kipps, T. J., Guadalupe, E., Chen, Y., Zhang, D., Wang, X., Davis, C.,. chronic lymphocytic leukaemia. Þ®ò Öò Ø¿»³¿¬±´ò ïèì: 605 615.. Signaling. ÚÛÞÍÖò 109.

(117) 18. Bresin, A., Cristofoletti, C., Caprini, E., Cantonetti, M., Monopoli, A., Russo, G. and Narducci, M. G. 2019. Preclinical Evidence for Targeting PI3K/mTOR. Cell Lymphoma. Öò ×²ª» ¬ò Ü»®³¿¬±´ò 19. Breuer, R., Becker, M. S., Brechmann, M., Mock, T., Arnold, R. and Krammer, P.. Öò Þ±´ò Ý¸»³ò îèç: 14996 15004. 20. Brinkmann, V., Billich, A., Baumruker, T., Heining, P., Schmouder, R., Francis, G., Aradhye, S. and Burtin, P. 2010. Fingolimod (FTY720): discovery and development of an oral drug to treat multiple sclerosis. Ò¿¬ò Î»ªò Ü®«¹ Ü ½±ªò ç: 883 897. 21. Broadhead, M. L., Clark, J. C. M., Myers, D. E., Dass, C. R. and Choong, P. F. M. 2011. The Molecular Pathogenesis of Osteosarcoma: A Review. Í¿®½±³¿. îðïï:. 22. Brugge, J., Hung, M.-C. and Mills, G. B. 2007. A new mutational AKTivation in the PI3K pathway. Ý¿²½»® Ý»´´. ïî: 104 107. 23. Cai, Q., Huang, H., Qian, D., Chen, K., Luo, J., Tian, Y., Lin, T. and Lin, T. 2013.. Ð´±ÍÑ²». è: e65308. 24. Camm, J., Hla, T., Bakshi, R. and Brinkmann, V. 2014. Cardiac and vascular ß³ò Ø»¿®¬ Öò ïêè: 632 644. 25. Casamayor, A., Morrice, N. A. and Alessi, D. R. 1999. Phosphorylation of Ser-241. 110.

(118) is essential for the activity of 3-phosphoinositide-dependent protein kinase-1: identification of five sites of phosphorylation in vivo. Þ ±½¸»³ò Öò íìî: 287 292. 26. Chen, L., Luo, L.-F., Lu, J., Li, L., Liu, Y.-F., Wang, J., Liu, H., Song, H., Jiang, H., Chen, S.-J., Luo, C. and Li, K. K. 2014. FTY720 induces apoptosis of M2. increasing endogenous ceramide levels. Ð´±ÍÑ²». ç: e103033.. W. C. 2004. Identification of specific PP2A complexes involved in human cell transformation. Ý¿²½»® Ý»´´. ë: 127 136. 28. Cho, U. S. and Xu, W. 2007. Crystal structure of a protein phosphatase 2A heterotrimeric holoenzyme. Ò¿¬«®». ììë: 53 57. 29. Christensen, D. J., Chen, Y., Oddo, J., Matta, K. M., Neil, J., Davis, E. D., Volkheimer, A. D., Lanasa, M. C., Friedman, D. R., Goodman, B. K., Gockerman,. 2011. SET oncoprotein overexpression in B-cell chronic lymphocytic leukemia and non-Hodgkin lymphoma: a predictor of aggressive disease and a new treatment target. Þ´±±¼. ïïè: 4150 4158. 30. Christensen, D. J., Chen, Y., Oddo, J., Matta, K. M., Neil, J., Davis, E. D., Volkheimer, A. D., Lanasa, M. C., Friedman, D. R., Goodman, B. K., Gockerman,. 2011. SET oncoprotein overexpression in B-cell chronic lymphocytic leukemia and non-Hodgkin lymphoma: a predictor of aggressive disease and a new treatment target. Þ´±±¼. ïïè: 4150 4158.. 111.

(119) Colton, C. A. and Vitek, M. P. 2011. Apolipoprotein E and peptide mimetics. phosphatase 2A. Öò ×³³«²±´ò Þ¿´¬ ³ò Ó¼ ïçëð. ïèê: 2535 2542. 32. Chun, J. and Hartung, H.-P. 2010. Mechanism of action of oral fingolimod (FTY720) in multiple sclerosis. Ý´ ²ò Ò»«®±°¸¿®³¿½±´ò íí: 91 101.. González-Alonso, P., Rojo, F. and García-Foncillas, J. 2017. Downregulation of. cancer. Ñ²½±¬¿®¹»¬. è: 40169 40180.. shows promising therapeutic implications and determines poor clinical outcome in patients with metastatic colorectal cancer. Ý´ ²ò Ý¿²½»® Î» ò Ñººò Öò ß³ò ß ±½ò Ý¿²½»® Î» ò îï: 347 356. 35. Dasari, S. and Tchounwou, P. B. 2014. Cisplatin in cancer therapy: molecular mechanisms of action. Û«®ò Öò Ð¸¿®³¿½±´ò ð: 364 378. 36. De Palma, R. M., Parnham, S. R., Li, Y., Oaks, J. J., Peterson, Y. K., Szulc, Z. M.,. the FTY720-SET complex reveals an alternative mechanism for the attenuation of ÚßÍÛÞ Öò Ñººò Ð«¾´ò Ú»¼ò ß³ò Í±½ò Û¨°ò Þ ±´ò íí: 7647 7666.. 112.

(120) chemosensitivity of leukemic cells to chemotherapeutic drugs. Ý»´´ò Í¹²¿´ò îé: 363 372.. and Huang, D. C. S. 2006. TheBH3 mimetic ABT-737 targets selective Bcl-2 proteins and efficiently induces apoptosis viaBak/Bax if Mcl-1 is neutralized. Ý¿²½»® Ý»´´. ïð: 389 99. 39. Deliconstantinos, G. and Tsakiris, S. 1985. Differential effect of anionic and cationic drugs on the synaptosome-associated acetylcholinesterase activity of dog Þ ±½¸»³ò Öò îîç: 81 86. 40. Eichhorn, P. J. A., Creyghton, M. P. and Bernards, R. 2009. Protein phosphatase Þ ±½¸ ³ò Þ ±°¸§ ò ß½¬¿ ÞÞß ó Î»ªò Ý¿²½»®. ïéçë: 1 15. 41. Elmore, S. 2007. Apoptosis: a review of programmed cell death. Ì±¨ ½±´ò Ð¿¬¸±´ò íë: 495 516.. T., Usui, T., Ohama, T. and Sato, K. 2015. The therapeutic effects of SET/I2PP2A Öò Ê»¬ò Ó»¼ò Í½ ò Ö°²ò Í±½ò Ê»¬ò Í½ ò. Takenouchi, H., Yoshino, S., Hazama, S., Nagano, H., Oshima, H., Oshima, M., Vitek, M. P., Matsuura, T., Hippo, Y., Usui, T., Ohama, T. and Sato, K. 2018. Ó±´ò Ý¿²½»® Î» ò ÓÝÎ. ïê: 554 563.. 113.

(121) suppressor NM23-H1 is a granzyme A-activated DNase during CTL-mediated Ý»´´. ïïî: 659 672. 45. Farrell, A. S., Allen-Petersen, B., Daniel, C. J., Wang, X., Wang, Z., Rodriguez, S., Impey, S., Oddo, J., Vitek, M. P., Lopez, C., Christensen, D. J., Sheppard, B. and. treatment of pancreatic cancer. Ó±´ò Ý¿²½»® Î» ò ÓÝÎ. ïî: 924 939. 46. Fernandes-Alnemri, T., Litwack, G. and Alnemri, E. S. 1994. CPP32, a novel. Öò Þ±´ò Ý¸»³ò îêç: 30761 30764. 47. Fernández, J. J., Candenas, M. L., Souto, M. L., Trujillo, M. M. and Norte, M. 2002. Okadaic acid, useful tool for studying cellular processes. Ý«®®ò Ó»¼ò Ý¸»³ò ç: 229 262. 48. Flegg, C. P., Sharma, M., Medina-Palazon, C., Jamieson, C., Galea, M., Brocardo, M. G., Mills, K. and Henderson, B. R. 2010. Nuclear Export and Centrosome Öò Þ±´ò Ý¸»³ò îèë: 18144 18154.. Sato, K. and Ohama, T. 2013. A potential therapeutic application of SET/I2PP2A Öò Ê»¬ò Ó»¼ò Í½ ò Ö°²ò Í±½ò Ê»¬ò Í½ ò éë: 349 354.. ECM-cell adhesion-independent drug resistance via PI3K-AKT and. 114.

(122) Ì«³±«® Þ±´ò Öò ×²¬ò Í±½ò Ñ²½±¼»ª»´±°³»²¬¿´ Þ ±´ò Ó»¼ò íí: 885 890. 51. García-Martínez, J. M. and Alessi, D. R. 2008. mTOR complex 2 (mTORC2). glucocorticoid-induced protein kinase 1 (SGK1). Þ ±½¸»³ò Öò ìïê: 375 385.. Patient-derived Human Non-small Cell Lung Cancer Cell Lines from a Primary Tumor and Corresponding Lymph Node Metastasis. ß²¬ ½¿²½»® Î» ò íê: 1507 1518.. Osteosarcoma. Ú®±²¬ò Ñ²½±´ò ì:. 54. Gunes, A., Scordo, M. G., Jaanson, P. and Dahl, M.-L. 2007. Serotonin and dopamine receptor genepolymorphisms and the risk of extrapyramidal side effects in perphenazine-treated schizophrenic patients. Ð §½¸±°¸¿®³¿½±´±¹§ øÞ»®´ò÷. ïçð: 479 484. 55. Gutierrez, A., Pan, L., Groen, R. W. J., Baleydier, F., Kentsis, A., Marineau, J.,. Weng, A. P., Yuan, H., Bradner, J. E., Mitsiades, C., Look, A. T. and Aster, J. C. 2014. Phenothiazines induce PP2A-mediated apoptosis in T cell acute Öò Ý´ ²ò ×²ª» ¬ò ïîì: 644 655.. 115.

(123) Amant, F., Waelkens, E., Sablina, A. and Janssens, V. 2016. Recurrent PPP2R1A Mutations in Uterine Cancer Act through a Dominant-Negative Mechanism to Promote Malignant Cell Growth. Ý¿²½»® Î» ò éê: 5719 5731. 57. Hahn, W. C., Dessain, S. K., Brooks, M. W., King, J. E., Elenbaas, B., Sabatini, D. M., DeCaprio, J. A. and Weinberg, R. A. 2002. Enumeration of the simian virus 40 early region elements necessary for human cell transformation. Ó±´ò Ý»´´ò Þ ±´ò îî: 2111 2123. 58. Haines, K. and Huang, G. S. 2019. Precision Therapy for Aggressive Endometrial Cancer by Reactivation of Protein Phosphatase 2A. Ý¿²½»® Î» ò éç: 4009 4010. 59. Hartung, B., Sampson, S. and Leucht, S. 2015. Perphenazine for schizophrenia. Ý±½¸®¿²»Ü¿¬¿¾¿ »Í§ ¬ò Î»ªò í: CD003443. 60. Henao, J. P., Peperzak, K. A., Lichvar, A. B., Orebaugh, S. L., Skledar, S. J., Pippi, M. A. and Williams, B. A. 2014. Extrapyramidal Symptoms Following Administration of Oral Perphenazine 4 or 8 mg An 11-Year Retrospective Analysis. Û«®ò Öò ß²¿» ¬¸» ±´ò íï: 231 235. 61. Henderson, M. C., Miranda, C. L., Stevens, J. F., Deinzer, M. L. and Buhler, D. R. 2000. In vitro inhibition of human P450 enzymes by prenylated flavonoids from hops, Humulus lupulus. È»²±¾ ±¬ ½¿ Ú¿¬»Ú±®» ¹² Ý±³°¼ò Þ±´ò Í§ ¬ò íð: 235 251. 62. Hino, H., Takaki, K. and Mochida, S. 2015. Inhibitor-1 and -2 of PP2A have preference between PP2A complexes. Þ ±½¸»³ò Þ ±°¸§ ò Î» ò Ý±³³«²ò ìêé: 297 302. 63. Ho, W. SC., Sizdahkhani, S., Hao, S., Song, H., Seldomridge, A., Tandle, A., Maric, D., Kramp, T., Lu, R., Heiss, J. D., Camphausen, K., Gilbert, M. R., Zhuang, Z. and Park, D. M. 2018. LB-100, anovel Protein Phosphatase 2A (PP2A). 116.

(124) Inhibitor, Sensitizes Malignant MeningiomaCells to the Therapeutic Effects of Radiation. Ý¿²½»® Ô»¬¬ò ìïë: 217 226. 64. Hu, X.-D., Liu, Y.-N., Zhang, Z.-Y., Ma, Z.-A., Suo, Z.-W. and Yang, X. 2015. Spinophilin-Targeted Protein Phosphatase-1 Alleviated Inflammatory Pain by Negative Control of MEK/ERK Signaling in Spinal Cord Dorsal Horn of Rats. Öò Ò»«®± ½ ò Ñººò Öò Í±½ò Ò»«®± ½ ò íë: 13989 14001. 65. Hu, X., Garcia, C., Fazli, L., Gleave, M., Vitek, M. P., Jansen, M., Christensen, D. and Mulholland, D. J. 2015. Inhibition of Pten deficient Castration Resistant Prostate Cancer by Targeting of the SET - PP2A Signaling axis. Í½ ò Î»°ò ë: 15182. 66. Huang, Y.-H., Chu, P.-Y., Chen, J.-L., Huang, C.-T., Lee, C.-H., Lau, K.-Y., Wang, W.-L., Wang, Y.-L., Lien, P.-J., Tseng, L.-M., Liu, C.-Y., Huang, Y.-H., Chu, P.-Y., Chen, J.-L., Huang, C.-T., Lee, C.-H., Lau, K.-Y., Wang, W.-L., Wang, Y.-L., Lien, P.-J., Tseng, L.-M. and Liu, C.-Y. 2018. SET Overexpression is Associated with Worse Recurrence-Free Survival in Patients with Primary Breast Cancer Receiving Adjuvant Tamoxifen Treatment. Öò Ý´ ²ò Ó»¼ò é: 245. 67. Ito, A., Koma, Y., Sohda, M., Watabe, K., Nagano, T., Misumi, Y., Nojima, H. and Kitamura, Y. 2003. Localization of thePP2A B56gammaregulatory subunit at the Golgi complex: possible role in vesicle transport and migration. ß³ò Öò Ð¿¬¸±´ò ïêî: 479 489. 68. Jacinto, E., Facchinetti, V., Liu, D., Soto, N., Wei, S., Jung, S. Y., Huang, Q., Qin, J. and Su, B. 2006. SIN1/MIP1 Maintains rictor-mTOR Complex Integrity and Regulates Akt Phosphorylation and Substrate Specificity. Ý»´´. ïîé: 125 137. 69. Janghorban, M., Farrell, A. S., Allen-Petersen, B. L., Pelz, C., Daniel, C. J., Oddo,. 117.

(125) J., Langer, E. M., Christensen, D. J. and Sears, R. C. 2014. Targeting c-MYC by antagonizing PP2A inhibitors in breast cancer. Ð®±½ò Ò¿¬´ò ß½¿¼ò Í½ ò Ëò Íò ßò ïïï: 9157 9162. 70. Janssens, V. and Goris, J. 2001. Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Þ ±½¸»³ò Öò íëí: 417 39. 71. Jiang, C.-H., Sun, T.-L., Xiang, D.-X., Wei, S.-S. and Li, W.-Q. 2018. Anticancer Activity and Mechanism of Xanthohumol: A Prenylated Flavonoid From Hops (Humulus lupulus L.). Ú®±²¬ò Ð¸¿®³¿½±´ò ç:. 72. Kake, S., Tsuji, S., Enjoji, S., Hanasaki, S., Hayase, H., Yabe, R., Tanaka, Y., Nakagawa, T., Liu, H.-P., Chang, S.-C., Usui, T., Ohama, T. and Sato, K. 2017. The roleof SET/I2PP2A in canine mammary tumors. Í½ ò Î»°ò é: 4279. 73. Kastrinsky, D. B., Sangodkar, J., Zaware, N., Izadmehr, S., Dhawan, N. S., Narla, G. and Ohlmeyer, M. 2015. Reengineered tricyclic anti-cancer agents. Þ ±±®¹ò Ó»¼ò Ý¸»³ò îí: 6528 6534. 74. Kauko, O., Imanishi, S. Y., Kulesskiy, E., Laajala, T. D., Yetukuri, L., Laine, A., Jumppanen, M., Haapaniemi, P., Ruan, L., Yadav, B., Suni, V., Varila, T., Corthals, G., Reimand, J., Wennerberg, K., Aittokallio, T. and Westermarck, J. 2018. Rules for PP2A-controlled phosphosignalling and drug responses. ¾ ±Î¨ ª. 271841. 75. Kauko, O. and Westermarck, J. 2018. Non-genomic mechanisms of protein phosphatase 2A (PP2A) regulation in cancer. ×²¬ò Öò Þ±½¸»³ò Ý»´´ Þ ±´ò çê: 157 164. 76. Kawahara, E., Maenaka, S., Shimada, E., Nishimura, Y. and Sakurai, H. 2013. Dynamic regulation of extracellular signal-regulated kinase (ERK) by protein. 118.

(126) Ð´±Í Ñ²». è: e63729. 77. Kawase, H., Okuwaki, M., Miyaji, M., Ohba, R., Handa, H., Ishimi, Y., Fujii-Nakata, T., Kikuchi, A. and Nagata, K. 1996. NAP-I is a functional homologue of TAF-I that is required for replication and transcription of the adenovirus genome in a chromatin-like structure. Ù»²» Ý»´´ Ü»ª±¬»¼ Ó±´ò Ý»´´ò Ó»½¸ò ï: 1045 1056. 78. Keyse, S. M. 2008. Dual-specificity MAPkinase phosphatases (MKPs) and cancer. Ý¿²½»® Ó»¬¿ ¬¿. Î»ªò îé: 253 261.. 79. Kockx, M., Guo, D. L., Traini, M., Gaus, K., Kay, J., Wimmer-Kleikamp, S., Rentero, C., Burnett, J. R., Le Goff, W., Van Eck, M., Stow, J. L., Jessup, W. and Kritharides, L. 2009. Cyclosporin A decreases apolipoprotein E secretion from human macrophages via a protein phosphatase 2B-dependent and ATP-binding cassette transporter A1 (ABCA1)-independent pathway. Öò Þ ±´ò Ý¸»³ò îèì: 24144 24154. 80. Kubota, D., Yoshida, A., Kawai, A. and Kondo, T. 2014. Proteomics identified overexpression of SET oncogene product and possible therapeutic utility of protein phosphatase 2A in alveolar soft part sarcoma. Öò Ð®±¬»±³»Î» ò ïí: 2250 2261. 81. Lai, D., Chen, M., Su, J., Liu, X., Rothe, K., Hu, K., Forrest, D. L., Eaves, C. J., Morin, G. B. and Jiang, X. 2018. PP2A inhibition sensitizes cancer stem cells to ABL tyrosine kinase inhibitors in BCR-ABL+ human leukemia. Í½ ò Ì®¿² ´ò Ó»¼ò ïð:. 82. Levine, B. and Yuan, J. 2005. Autophagy in cell death: an innocent convict? Öò Ý´ ²ò ×²ª» ¬ò ïïë: 2679 2688. 119.

(127) 83. Li, F.-Q., Sempowski, G. D., McKenna, S. E., Laskowitz, D. T., Colton, C. A. and Vitek, M. P. 2006. Apolipoprotein E-derived peptides ameliorate clinical disability and inflammatory infiltrates into the spinal cord in a murine model of multiple sclerosis. Öò Ð¸¿®³¿½±´ò Û¨°ò Ì¸»®ò íïè: 956 965. 84. Li, M., Guo, H. and Damuni, Z. 1995. Purification and characterization of two potent heat-stable protein inhibitors of protein phosphatase 2A from bovine kidney. Þ ±½¸»³ ¬®§. íì: 1988 1996. 85. Lindern, M. von, Baal, S. van, Wiegant, J., Raap, A., Hagemeijer, A. and Grosveld, G. 1992. Can, a putative oncogene associated with myeloid leukemogenesis, may be activated by fusion of its 3 half to different genes: characterization of the set gene. Ó±´ò Ý»´´ò Þ ±´ò ïî: 3346 3355. 86. Linnet, K. and Wiborg, O. 1996. Steady-state serum concentrations of the neuroleptic perphenazine in relation to CYP2D6 genetic polymorphism. Ý´ ²ò Ð¸¿®³¿½±´ò Ì¸»®ò êð: 41 47. 87. Little, A. S., Smith, P. D. and Cook, S. J. 2013. Mechanisms of acquired resistance to ERK1/2 pathway inhibitors. Ñ²½±¹»²». íî: 1207 1215. 88. Liu, H., Gu, Y., Wang, H., Yin, J., Zheng, G., Zhang, Z., Lu, M., Wang, C. and He, Z. 2015. Overexpression of PP2A inhibitor SET oncoprotein is associated with tumor progression and poor prognosis in human non-small cell lung cancer. Ñ²½±¬¿®¹»¬. ê: 14913 14925. 89. Liu, T., Zhou, Y., Ko, K. S. and Yang, H. 2015. Interactions between Myc and Mediators of Inflammation in Chronic Liver Diseases. Ó»¼ ¿¬±® ×²º´¿³³ò îðïë: 276850. 90. Lu, J., Kovach, J. S., Johnson, F., Chiang, J., Hodes, R., Lonser, R. and Zhuang, Z.. 120.

(128) 2009. Inhibition of serine/threonine phosphatase PP2A enhances cancer chemotherapy by blocking DNA damage induced defense mechanisms. Ð®±½ò Ò¿¬´ò ß½¿¼ò Í½ ò Ëò Íò ßò ïðê: 11697 11702. 91. McClinch, K., Avelar, R. A., Callejas, D., Izadmehr, S., Wiredja, D., Perl, A., Sangodkar, J., Kastrinsky, D. B., Schlatzer, D., Cooper, M., Kiselar, J., Stachnik, A., Yao, S., Hoon, D., McQuaid, D., Zaware, N., Gong, Y., Brautigan, D. L., Plymate, S. R., Sprenger, C. C. T., Oh, W. K., Levine, A. C., Kirschenbaum, A., Sfakianos, J. P., Sears, R., DiFeo, A., Ioannou, Y., Ohlmeyer, M., Narla, G. and Galsky, M. D. 2018. Small-MoleculeActivators of Protein Phosphatase 2A for the Treatment of Castration-Resistant Prostate Cancer. Ý¿²½»® Î» ò éè: 2065 2080. 92. McCright, B., Rivers, A. M., Audlin, S. and Virshup, D. M. 1996. The B56 Family of Protein Phosphatase 2A (PP2A) Regulatory Subunits Encodes Differentiation-induced Phosphoproteins That Target PP2A to Both Nucleus and Cytoplasm. Öò Þ ±´ò Ý¸»³ò îéï: 22081 22089. 93. McCubrey, J. A., Steelman, L. S., Chappell, W. H., Abrams, S. L., Wong, E. WT., Chang, F., Lehmann, B., Terrian, D. M., Milella, M., Tafuri, A., Stivala, F., Libra, M., Basecke, J., Evangelisti, C., Martelli, A. M. and Franklin, R. A. 2007. ROLES OF THE RAF/MEK/ERK PATHWAY IN CELL GROWTH, MALIGNANT TRANSFORMATION AND DRUG RESISTANCE. Þ ±½¸ ³ò Þ±°¸§ ò ß½¬¿. ïééí: 1263 1284. 94. Miller, A. V, Hicks, M. a, Nakajima, W., Richardson, A. C., Windle, J. J. and Harada, H. 2013. Paclitaxel-induced apoptosis is BAK-dependent, but BAX and BIM-independent in breast tumor. Ð´±ÍÑ²». è: e60685. 95. Miyaji-Yamaguchi, M., Okuwaki, M. and Nagata, K. 1999. Coiled-coil. 121.

(129) structure-mediated dimerization of template activating factor-I is critical for its chromatin remodeling activity11Edited by T. Richmond. Öò Ó±´ò Þ±´ò îçð: 547 557. 96. Mody, H. R., Hung, S. W., Naidu, K., Lee, H., Gilbert, C. A., Hoang, T. T., Pathak, R. K., Manoharan, R., Muruganandan, S. and Govindarajan, R. 2017. SET contributes to the epithelial-mesenchymal transition of pancreatic cancer. Ñ²½±¬¿®¹»¬. è: 67966 67979. 97. Molina, G., Vogt, A., Bakan, A., Dai, W., de Oliveira, P. Q., Znosko, W., Smithgall, T. E., Bahar, I., Lazo, J. S., Day, B. W. and Tsang, M. 2009. Zebrafish chemical screening reveals an inhibitor of Dusp6 that expands cardiac cell lineages. Ò¿¬ò Ý¸»³ò Þ±´ò ë: 680 687. 98. Neviani, P., Harb, J. G., Oaks, J. J., Santhanam, R., Walker, C. J., Ellis, J. J., Ferenchak, G., Dorrance, A. M., Paisie, C. A., Eiring, A. M., Ma, Y., Mao, H. C., Zhang, B., Wunderlich, M., May, P. C., Sun, C., Saddoughi, S. A., Bielawski, J., Blum, W., Klisovic, R. B., Solt, J. A., Byrd, J. C., Volinia, S., Cortes, J., Huettner, C. S., Koschmieder, S., Holyoake, T. L., Devine, S., Caligiuri, M. A., Croce, C. M., Garzon, R., Ogretmen, B., Arlinghaus, R. B., Chen, C.-S., Bittman, R., Hokland, P., Roy, D.-C., Milojkovic, D., Apperley, J., Goldman, J. M., Reid, A., Mulloy, J. C., Bhatia, R., Marcucci, G. and Perrotti, D. 2013. PP2A-activating drugs selectively eradicate TKI-resistant chronic myeloid leukemic stem cells. Öò Ý´ ²ò ×²ª» ¬ò ïîí: 4144 4157. 99. Neviani, P. and Perrotti, D. 2014. SETting OP449 into the PP2A-Activating Drug Family. Ý´ ²ò Ý¿²½»® Î» ò îð: 2026 2028. 100. Neviani, P., Santhanam, R., Trotta, R., Notari, M., Blaser, B. W., Liu, S., Mao, H.,. 122.

(130) Chang, J. S., Galietta, A., Uttam, A., Roy, D. C., Valtieri, M., Bruner-Klisovic, R., Caligiuri, M. A., Bloomfield, C. D., Marcucci, G. and Perrotti, D. 2005. The tumor suppressor PP2A is functionally inactivated in blast crisis CML through the inhibitory activity of the BCR/ABL-regulated SET protein. Ý¿²½»® Ý»´´. è: 355 368. 101. Niba, E. T. E., Nagaya, H., Kanno, T., Tsuchiya, A., Gotoh, A., Tabata, C., Kuribayashi, K., Nakano, T. and Nishizaki, T. 2013. Crosstalk between PI3 kinase/PDK1/Akt/Rac1 and Ras/Raf/MEK/ERK pathways downstream PDGF receptor. Ý»´´ò Ð¸§ ±´ò Þ±½¸»³ò ×²¬ò Öò Û¨°ò Ý»´´ò Ð¸§ ±´ò Þ ±½¸»³ò Ð¸¿®³¿½±´ò íï: 905 913. 102. Nishi, A., Kuroiwa, M. and Shuto, T. 2011. Mechanisms for the modulation of dopamine D1 receptor signaling in striatal neurons. Ú®±²¬ò Ò»«®±¿²¿¬ò ë: 43. 103. Oaks, J. J., Santhanam, R., Walker, C. J., Roof, S., Harb, J. G., Ferenchak, G., Eisfeld, A.-K., Van Brocklyn, J. R., Briesewitz, R., Saddoughi, S. A., Nagata, K., Bittman, R., Caligiuri, M. A., Abdel-Wahab, O., Levine, R., Arlinghaus, R. B., Quintas-Cardama, A., Goldman, J. M., Apperley, J., Reid, A., Milojkovic, D., Ziolo, M. T., Marcucci, G., Ogretmen, B., Neviani, P. and Perrotti, D. 2013. Antagonistic activities of the immunomodulator and PP2A-activating drug FTY720 (Fingolimod, Gilenya) in Jak2-driven hematologic malignancies. Þ´±±¼. ïîî: 1923 1934. 104. Oaks, J. and Ogretmen, B. 2014. Regulation of PP2A by Sphingolipid Metabolism and Signaling. Ú®±²¬ò Ñ²½±´ò ì: 388. 105. Omar, H. A., Chou, C.-C., Berman-Booty, L. D., Ma, Y., Hung, J.-H., Wang, D., Kogure, T., Patel, T., Terracciano, L., Muthusamy, N., Byrd, J. C., Kulp, S. K. and. 123.

(131) Chen, C.-S. 2011. Antitumor Effects of OSU-2S, a Non-immunosuppressive Analogue of FTY720, in Hepatocellular Carcinoma. Ø»°¿¬±´ò Þ¿´¬ ³ò Ó¼. ëí: 1943 1958. 106. Otálora-Otálora, B. A., Henríquez, B., López-Kleine, L. and Rojas, A. 2019. RUNX family: Oncogenes or tumor suppressors (Review). Ñ²½±´ò Î»°ò ìî: 3 19. 107. Ozdemir, E., Bagcivan, I. and Gursoy, S. 2013. Role of D1/D2 dopamin receptors antagonist perphenazine in morphine analgesia and tolerance in rats. Þ± ²ò Öò Þ¿ ½ Ó»¼ò Í½ ò ïí: 119 125. 108. Pellicena, P. and Schulman, H. 2014. CaMKII inhibitors: from research tools to therapeutic agents. Ú®±²¬ò Ð¸¿®³¿½±´ò ë:. 109. Pippa, R., Dominguez, A., Malumbres, R., Endo, A., Arriazu, E., Marcotegui, N., Guruceaga, E. and Odero, M. D. 2017. MYC-dependent recruitment of RUNX1 and GATA2 on the SET oncogene promoter enhances PP2A inactivation in acute myeloid leukemia. Ñ²½±¬¿®¹»¬. è: 53989 54003. 110. Puustinen, P., Junttila, M. R., Vanhatupa, S., Sablina, A. A., Hector, M. E., Teittinen, K., Raheem, O., Ketola, K., Lin, S., Kast, J., Haapasalo, H., Hahn, W. C. and Westermarck, J. 2009. PME-1 protects extracellular signal-regulated kinase pathway activity from protein phosphatase 2A-mediated inactivation in human malignant glioma. Ý¿²½»® Î» ò êç: 2870 2877. 111. Querfeld, C., Rizvi, M. A., Kuzel, T. M., Guitart, J., Rademaker, A., Sabharwal, S.. Enzastaurin Induces Apoptosis in Cutaneous T-Cell Lymphoma Cell Lines through the AKT Pathway. Öò ×²ª» ¬ò Ü»®³¿¬±´ò ïîê: 1641 1647. 112. Rangel-Barajas, C., Coronel, I. and Florán, B. 2015. Dopamine Receptors and. 124.