Title
Analysis of Primary and Metastatic Tumor in Fluorescent Protein
Expressing Metastasis Mouse Model : (1) Imaging
Nuclear-Cytoplasmic Dynamics in Primary and Metastatic Colon Cancer
in Nude Mice(2) Imaging the Role of Multinucleate Pancreatic
Cancer Cells and Cancer-Associated Fibroblasts in Peritoneal
Metastasis in Mouse Models( 内容と審査の要旨(Summary) )
Author(s)
長谷川, 恒輔
Report No.(Doctoral
Degree)
博士(医学) 甲第1067号
Issue Date
2018-03-25
Type
博士論文
Version
none
URL
http://hdl.handle.net/20.500.12099/75204
※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。Ặྡ㸦ᮏ⡠㸧 Ꮫ ࡢ ✀ 㢮 Ꮫᤵ␒ྕ Ꮫᤵ᪥ Ꮫᤵせ௳ Ꮫㄽᩥ㢟┠ ᑂ ᰝ ጤ ဨ 㛗 ㇂ ᕝ ᜏ ㍜㸦ᒸᒣ┴㸧 ༤ ኈ㸦་Ꮫ㸧 ⏥➨ ྕ ᖹᡂ ᖺ ᭶ ᪥ Ꮫつ๎➨㸲᮲➨㸯㡯ヱᙜ
$QDO\VLV RI 3ULPDU\ DQG 0HWDVWDWLF 7XPRU LQ )OXRUHVFHQW 3URWHLQ ([SUHVVLQJ0HWDVWDVLV0RXVH0RGHO
,PDJLQJ 1XFOHDU&\WRSODVPLF '\QDPLFV LQ 3ULPDU\ DQG 0HWDVWDWLF &RORQ &DQFHULQ1XGH0LFH ,PDJLQJWKH5ROHRI0XOWLQXFOHDWH3DQFUHDWLF&DQFHU&HOOVDQG&DQFHU $VVRFLDWHG)LEUREODVWVLQ3HULWRQHDO0HWDVWDVLVLQ0RXVH0RGHOV 㸦ᰝ㸧ᩍᤵ ἑ Ẏ 㸦ᰝ㸧ᩍᤵ ྜྷ⏣ ᘯ ᩍᤵ ᒣཱྀ ▐ ㄽ ᩥ ෆ ᐜ ࡢ せ ᪨ ⒴ࡢ㌿⛣࣭ᾐ₶ᛶࡣᝏᛶ⭘⒆ᛶᝈࡢணᚋࢆつᐃࡍࡿせ࡞ᅉᏊ࡛࠶ࡿࠋ⭘⒆ࡢ㐍ᒎࡣ㸪⒴⣽⬊ࡑ ࡢࡶࡢࡢᛶ㉁ຍ࠼㸪⒴⣽⬊ࡢ࿘ᅖࢆࡾࡲࡃ⒴ᚤᑠ⎔ቃ㸦7XPRU 0LFURHQYLURQPHQW 70(㸧ࡀ㔜せ ࡞ᙺࢆᯝࡓࡋ࡚࠸ࡿࠋ≉㸪⒴㛵㐃⥺⥔ⱆ⣽⬊㸦&DQFHU$VVRFLDWHG)LEUREODVW &$)㸧ࡸࣜࣥࣃ ⌫ᶞ≧⣽⬊࣐ࢡࣟࣇ࣮ࢪ࡞ 70( ࢆᵓᡂࡍࡿ⣽⬊ࡣ㸪⒴⣽⬊ࡢ┦స⏝ࢆ㏻ࡋ࡚⒴ࡢ㌿⛣ᛶࡸ ᾐ₶ᛶࡢไᚚ῝ࡃ㛵ࡋ࡚࠸ࡿࡇࡀ᫂ࡽࡉࢀ࡚࠸ࡿࠋࡋࡋ㸪⏕యෆ࠾࠸࡚ᐇ㝿ࡇࡢࡼ ࠺࡞┦స⏝ࢆほᐹࡍࡿࡇࡣᅔ㞴࡛࠶ࡾ㸪ࡑࡢヲ⣽࡞ᶵᗎࡘ࠸࡚ࡣ༑ศ⌮ゎࡉࢀ࡚࠸࡞࠸ࠋࡑ ࡇ࡛㸪ᮏ◊✲࡛ࡣ㸪⭘⒆㌿⛣ᕢ࠾ࡅࡿ⒴⣽⬊ 70( ࢆᵓᡂࡍࡿ⣽⬊ࡢ┦స⏝ࢆྍどࡍࡿࡇ ࡼࡗ࡚㸪ࡑࢀࡒࢀࡢ⣽⬊ࡢືែࢆほᐹࡋゎᯒࢆ⾜࠺ࡓࡵࡢࣔࢹࣝ⣔ࢆᵓ⠏ࡍࡿࡇࢆ┠ᶆࡋ㸪ࡇ ࡢࡓࡵࡢ➨୍Ṍࡋ࡚௨ୗࡢ᳨ウࢆ⾜ࡗࡓࠋ㸦㸧⥳Ⰽ⺯ගࢱࣥࣃࢡ㉁㸦*UHHQ)OXRUHVFHQW3URWHLQ *)3㸧ࣄࢫࢺࣥ㸦+%㸧ࡢ⼥ྜࢱࣥࣃࢡ㉁㸦*)3+%㸧ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊ࠾ࡼࡧ㉥Ⰽ⺯ගࢱࣥࣃ ࢡ㉁㸦5HG)OXRUHVFHQW3URWHLQ 5)3㸧ࢆࢥ࣮ࢻࡍࡿ㑇ఏᏊࢆඹᑟධࡋࡓ⭠⒴ᰴࢆࢾ࣮ࢻ࣐࢘ࢫ ⛣᳜ࡍࡿࡇࡼࡾ⭠⒴㌿⛣ࣔࢹࣝࢆసᡂࡋཎⓎᕢ㌿⛣ᕢࡢ⒴⣽⬊ࡢᙧែࡢ㐪࠸ࢆㄪࡓࠋ㸦㸧 *)3 㑇ఏᏊࢆᑟධࡋࡓ⮅⮚⒴ᰴࢆࢾ࣮ࢻ࣐࢘ࢫ⛣᳜ࡍࡿࡇࡼࡾ⮅⮚⒴㌿⛣ࣔࢹࣝࢆసᡂࡋཎ Ⓨᕢ㌿⛣ᕢࢆᵓᡂࡍࡿ⒴⣽⬊ࡢᙧែቑṪ⬟ࡢ㐪࠸ࢆゎᯒࡋࡓࠋ ࠙ᑐ㇟᪉ἲࠚ ձ ࣄࢺ⤖⭠⭢⒴ᰴ㸦+&7㸧ᑐࡋࣞࢺࣟ࢘ࣝࢫ࣋ࢡࢱ࣮ࢆ⏝࠸࡚*)3+%࠾ࡼࡧ 5)3㑇ఏᏊࢆ ᑟධࡋ㸪⣽⬊㉁࠾ࡼࡧᰁⰍయࡀࡑࢀࡒࢀ 5)3 ࠾ࡼࡧ *)3 ࡼࡗ࡚ᶆ㆑ࡉࢀࡓ⣽⬊ᰴ㸦+&7*)3 5)3 ⣽⬊㸧ࢆᚓࡓࠋ+&7*)35)3 ⣽⬊ࢆ 㐌㱋ࡢࢾ࣮ࢻ࣐࢘ࢫ %$/%FQXQX ࡢ⭁⮚ෆ⛣᳜ ࡋ⭠⒴㌿⛣ࣔࢹࣝࢆసᡂࡋࡓࠋ⛣᳜ᚋ ᪥┠࣐࢘ࢫࢆゎ๗ࡋ ཎⓎᕢ㌿⛣ᕢࡢ⭘⒆ࡘ ࠸࡚H[ YLYR࣓࣮ࢪࣥࢢࡼࡗ࡚⒴⣽⬊ࡢᙧែࡸቑṪᵝᘧࢆほᐹࡋࡓࠋࡉࡽ㸪ཎⓎᕢ㌿⛣ ᕢࡢ⭘⒆ሢࡽ⒴⣽⬊ࢆ᥇ྲྀࡋࢥࣥࣇ࢛࣮࢝ࣝ㢧ᚤ㙾ࡼࡗ࡚⣽⬊ࡢᙧែࢆほᐹࡋࡓࠋ ղ ࣄࢺ⮅⮚⭢⒴ᰴ㸦%[3&㸧ᑐࡋࣞࢺࣟ࢘ࣝࢫ࣋ࢡࢱ࣮ࢆ⏝࠸࡚*)3㑇ఏᏊࢆᑟධࡋ㸪*)3 ࢱࣥ
ࣃࢡ㉁ࡼࡗ࡚ᶆ㆑ࡉࢀࡓ %[3& ⣽⬊ᰴ㸦%[3&*)3 ⣽⬊㸧ࢆᚓࡓࠋࡲࡎ㸪%[3&*)3 ⣽⬊ࢆࢾ࣮
ࢻ࣐࢘ࢫࡢ⓶ୗ⛣᳜ࡋ⓶ୗ⭘⒆ࢆᙧᡂࡉࡏࡓࠋḟ㸪ࡇࡢ⓶ୗ⭘⒆ࢆฟࡋࡓᚋ㸪PPࡢ⭘⒆ ∦ศࡋูࡢࢾ࣮ࢻ࣐࢘ࢫࡢ⮅⮚ᑿ㒊⛣᳜ࡋࡓࠋ⛣᳜ᚋ㸪 㐌┠࣐࢘ࢫࢆ㛤⭡ࡋཎⓎ ᕢ㌿⛣ᕢࡢ⭘⒆ࡘ࠸࡚H[ YLYR࣓࣮ࢪࣥࢢࢆ⾜࠸⒴⣽⬊ࡢᙧែࢆほᐹࡋࡓࠋࡲࡓ㸪ࡑࢀࡒ ࢀࡢ⭘⒆ࢆศᩓࡉࡏ⣽⬊ᇵ㣴ࢆ⾜࠸㸪⣽⬊ࡢᙧែࢆゎᯒࡋࡓࠋࡉࡽ㸪ཎⓎᕢ࠾ࡼࡧ㌿⛣ᕢࡢ⒴ ⣽⬊ࡢ⭘⒆ᙧᡂ⬟ࡢᕪ␗ࢆㄪࡿࡓࡵ㸪ࡑࢀࡒࢀࡢ⣽⬊ࢆࢾ࣮ࢻ࣐࢘ࢫࡢ⓶ୗ⛣᳜ࡋ㸪⓶ୗ ⭘⒆ࢆᙧᡂࡉࡏࡓࠋ⛣᳜ᚋ ᪥┠⓶ୗ⭘⒆ࢆฟࡋ⭘⒆ᚄ㔜㔞ࢆ ᐃࡋࡓࠋ
࠙⤖ᯝࠚ ձ +&7*)35)3 ⣽⬊ࢆ⭁⮚⛣᳜ࡋࡓ࣐࢘ࢫࡘ࠸࡚㸪⛣᳜ᚋ ᪥┠ゎ๗ࢆ⾜ࡗࡓࡇࢁ㸪⫢ ⮚࠾ࡼࡧᚋ⭡⭷⭘⒆ࡢ㌿⛣ᕢࡀㄆࡵࡽࢀࡓࠋࡇࢀࡽ㌿⛣ᕢࢆ⺯ග࣓࣮ࢪࣥࢢࢩࢫࢸ࣒ࡼࡗ ࡚ほᐹࡋࡓࡇࢁ㸪⭘⒆୰ *)3 ࠾ࡼࡧ 5)3 ࡢ⺯ගࡼࡗ࡚ᶆ㆑ࡉࢀࡓ⒴⣽⬊ࡀㄆࡵࡽࢀಶࠎࡢ ⒴⣽⬊ࡢᙧែࡸ⣽⬊ቑṪࡢ≧ែࢆほᐹࡍࡿࡇࡀ࡛ࡁࡓࠋࡲࡓ㸪ཎⓎᕢ࠾ࡼࡧ㌿⛣ᕢࡽᚓࡽࢀ ࡓᇵ㣴⒴⣽⬊ࢆほᐹࡋࡓࡇࢁ㸪㌿⛣ᕢࡽᚓࡽࢀࡓ⣽⬊ࡘ࠸࡚⣽⬊㉁ෆ✵⬊ࢆᣢࡘከ᰾ࡢ ⣽⬊ࡸ⣳㗽≧ࡢ⣽⬊ࡀㄆࡵࡽࢀࡓࠋࡇࢀࡽࡢ␗ᖖ࡞ᙧែࢆ♧ࡍ⣽⬊ࡣ㸪㏻ᖖࡢᇵ㣴ࢆ⾜ࡗࡓ +&7 ⣽⬊ࡢ୰ࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ ղ %[3&*)3 ⣽⬊⓶ୗ⭘⒆ࢆ⮅⮚ᑿ㒊⛣᳜ࡋࡓ࣐࢘ࢫࡘ࠸࡚㸪⛣᳜ᚋ 㐌ᚋゎ๗ࢆ⾜ࡗࡓ ࡇࢁ㸪⛣᳜㒊㸦ཎⓎᕢ㸧࠾ࡼࡧ⭡⭷ୖ㸦㌿⛣ᕢ㸧⭘⒆ࡢᙧᡂࡀㄆࡵࡽࢀࡓࠋཎⓎᕢ࠾ࡼࡧ㌿⛣ᕢ ࡽ᥇ྲྀࡋࡓ⣽⬊ࢆᇵ㣴ࡋࡓࡇࢁ㌿⛣⭘⒆ሢࡽᚓࡽࢀࡓ⣽⬊ࡢ୰ከᩘࡢከ᰾⒴⣽⬊ࣞ ࢩࣆ࢚ࣥࢺ࣐࢘ࢫ⏤᮶ࡢ⥺⥔ⱆ⣽⬊ࡀᏑᅾࡍࡿࡇࡀࢃࡗࡓࠋࡲࡓ㸪ཎⓎᕢ࠾ࡼࡧ㌿⛣ᕢ⏤᮶ ࡢ⣽⬊ࡢ⓶ୗ⭘⒆ᙧᡂ⬟ࢆㄪࡓࡇࢁ㌿⛣ᕢ⏤᮶ࡢ⣽⬊ࡼࡗ࡚ᙧᡂࡉࢀࡓ⓶ୗ⭘⒆ࡣ㸪ཎⓎ ᕢ⏤᮶ࡢ⭘⒆ẚ㍑ࡋ࡚㸪᭷ព⭘⒆㔜㔞ࡀቑຍࡋ࡚࠸ࡓࠋ ࠙⪃ᐹࠚ ⺯ගࢱࣥࣃࢡ㉁ࡼࡗ࡚ᶆ㆑ࡉࢀࡓࣄࢺ⭠⒴⣽⬊࠾ࡼࡧ⮅⮚⒴⣽⬊ࢆࢾ࣮ࢻ࣐࢘ࢫ⛣᳜ࡍࡿࡇ ࡼࡾ⭘⒆㌿⛣ࣔࢹࣝࢆసᡂࡋࡓࠋࡇࡢࡼ࠺࡞⭘⒆㌿⛣ࣔࢹ࣐ࣝ࢘ࢫࢆ⏝࠸࡚㸪ᙧᡂࡉࢀࡓ⭘⒆ࢆ ⺯ග࣓࣮ࢪࣥࢢࢩࢫࢸ࣒ࡼࡗ࡚ほᐹࡋࡓࡇࢁ㸪⭘⒆ෆ࠾ࡅࡿಶࠎࡢ⒴⣽⬊ࡢືែࢆ⺯ගࡼ ࡗ࡚ࣜࣝࢱ࣒ࣔࢽࢱࣜࣥࢢ࡛ࡁࡿࡇࡀࢃࡗࡓࠋ㌿⛣⭘⒆ᕢࡢ⒴⣽⬊ࡢᙧែࢆほᐹࡋࡓࡇ ࢁ㸪㌿⛣ᕢࡣከᩘࡢከ᰾⒴⣽⬊ࡀᏑᅾࡍࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋከࡃࡢ⒴⣽⬊࡛ࡣᝏᛶࡢ㐍ᒎ క࠸ᰁⰍయࡢಸຍࡀ㉳ࡇࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࡀ㸪ࡇࡢࡼ࠺࡞ᰁⰍయࡢಸຍࡀ⣽⬊᰾ࡢከ᰾ࢆ⤒ ࡚ᘬࡁ㉳ࡇࡉࢀࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋࡲࡓ㸪⮅⮚⒴㌿⛣⭘⒆ᕢࡽᚓࡽࢀࡓ⣽⬊ࡢ୰࣐࢘ࢫ⏤᮶ ࡢከᩘࡢ⥺⥔ⱆ⣽⬊ࡀΰᅾࡋ࡚࠸ࡿࡇࡀࢃࡗࡓࠋ⮅⮚⒴㌿⛣⭘⒆ᕢࡽᚓࡽࢀࡓ⣽⬊ࡣཎⓎᕢ ࡽᚓࡽࢀࡓ⣽⬊ẚ㍑ࡋ࡚㧗࠸⭘⒆ᙧᡂ⬟ࢆᣢࡘࡇࡼࡾ㸪࣐࢘ࢫࡢ⥺⥔ⱆ⣽⬊ࡀ &$) ࡋ࡚స⏝ࡍ ࡿࡇࡼࡗ࡚⭘⒆ࡢ㐍ᒎࢆಁࡍᙺࢆᯝࡓࡋ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ ࠙⤖ㄽࠚ ⺯ගࢱࣥࣃࢡ㉁ࡼࡗ࡚ᶆ㆑ࡉࢀࡓ⭠⒴⣽⬊࠾ࡼࡧ⮅⮚⒴⣽⬊ࢆ⏝࠸࡚⭘⒆㌿⛣ࣔࢹࣝࢆసᡂࡍ ࡿࡇࡼࡗ࡚㌿⛣ᕢࢆᵓᡂࡍࡿ⒴⣽⬊ࡢ⣽⬊ືែࢆ㠀くⓗほᐹࡍࡿࡓࡵࡢࢩࢫࢸ࣒ࢆᵓ⠏ࡋ ࡓࠋ ㄽ ᩥ ᑂ ᰝ ࡢ ⤖ ᯝ ࡢ せ ᪨ ⏦ㄳ⪅ 㛗㇂ᕝᜏ㍜ࡣ㸪⺯ගࢱࣥࣃࢡ㉁ࡼࡗ࡚ᶆ㆑ࡉࢀࡓࣄࢺ⒴⣽⬊ࢆࢾ࣮ࢻ࣐࢘ࢫ⛣᳜ࡍࡿࡇ ࡼࡗ࡚⭘⒆㌿⛣ࣔࢹࣝࢆసᡂࡋ㸪H[YLYR⺯ග࣓࣮ࢪࣥࢢࢩࢫࢸ࣒ࢆ⏝࠸࡚㌿⛣⭘⒆ᕢ࠾ࡅ ࡿ⒴⣽⬊ࡢ⣽⬊ືែࢆほᐹࡍࡿᡭἲࢆᵓ⠏ࡋࡓࠋᮏ◊✲ᡂᯝࡣ㸪⒴ࡢ㌿⛣ࡸᝏᛶࡢ⤌ࡳࢆゎᯒࡍ ࡿࡓࡵࡢᇶ┙ᢏ⾡ࢆᥦ౪ࡍࡿࡶࡢ࡛࠶ࡾ㸪ᚋࡢ⒴◊✲ࡢⓎᒎᑡ࡞ࡽࡎᐤࡍࡿ⪃࠼ࡽࢀࡿࠋ 㹙ㄽᩥබ⾲ㄅ㹛 㸧.268.(+$6(*$:$$7686+,68(768*80,.,1$.$085$7$.8520$7680272+,720,$2.,7$.$+,52 .81,6$'$ 0,&+$(/ %289(7 0$6$+,72 6+,0,=8 DQG 52%(57 0 +2))0$1 ,PDJLQJ 1XFOHDU &\WRSODVPLF'\QDPLFVLQ3ULPDU\DQG0HWDVWDWLF&RORQ&DQFHULQ1XGH0LFH $QWLFDQFHU5HVHDUFK 㸧.268.(+$6(*$:$$7686+,68(768*80,.,1$.$085$7$.8520$7680272+,720,$2.,7$.$+,52 .81,6$'$0$6$+,726+,0,=86+,*(72<26$-,+,6$7$.$025,:$.,DQG52%(570+2))0$1 ,PDJLQJ WKH 5ROH RI 0XOWLQXFOHDWH 3DQFUHDWLF &DQFHU &HOOV DQG &DQFHU$VVRFLDWHG )LEUREODVWV LQ 3HULWRQHDO0HWDVWDVLVLQ0RXVH0RGHOV
