Depar t ment of Obs t et r i cs and Gy necol ogy
Tadao Tanaka,Professor Kazunori Ochiai,Professor Makoto Yasuda,Professor Kazuhiko Ochiai,Professor Hiroshi Sasaki,Professor Naoki Kamiya,Professor Takekazu Onda,Associate Professor Seiji Isonishi,Associate Professor Shigeki Niimi,Associate Professor Shigemitu Kobayashi,Assistant Professor Aikou Okamoto,Assistant Professor Kyousuke Yamada,Assistant Professor Kuniaki Ohura,Assistant Professor, Hirokuni Takano,Assistant Professor Satoshi Takakura,Assistant Professor
Gener al Summar y
The ma i n r e s e a r c h t opi c s of our de pa r t me nt a r e t he de v e l opme nt of mol e c ul a r l y t a r g e t e d a g e nt s f or g y ne c ol og i c t umor s ,i nc l udi ng ov a r i a n c a nc e r ,c l a r i f i c a t i on of t he me c ha ni s ms of s uc c e s s f ul pr e g na nc y ,a nd t he de v e l opme nt of a s s i s t e d r e pr oduc t i v e t e c hni que s . The s e t opi c s we r e i nv e s t i g a t e d bot h e x pe r i me nt a l l y a nd c l i ni c a l l y .
Res ear ch Act i v i t i es Gy ne c o l o g i c o nc o l o g y
1 . CD1 4 7( e x t r a c e l l ul a r ma t r i x me t a l l opr ot e a s e i nduc e r , ba s i g i n)pl a y s i mpor t a nt r ol e s i n t umor i nv a s i v e ne s s a nd me t a s t a s i s by a c t i v a t i on of ma t r i x me t a l l opr ot e i na s e s ( MMPs ) . We s howe d t ha t CD1 4 7 wa s e x pr e s s e d i n 9 7 . 5 %of e ndome t r i a l c a r c i noma s a nd i n 8 3 . 1 % of c e r v i c a l s qua mous c e l l c a r c i noma s . Es pe c i a l l y i n e ndome t r i a l c a r - c i noma s ,CD1 4 7 e x pr e s s i on i s c or r e l a t e d wi t h v a r i ous c l i ni c opa t hol og i c a l f a c t or s( s t a g e , g r a de , l y mph node me t a s t a s i s , l y mpha t i c v e s s e l i nf i l t r a t i on, di s t a nt me t a s t a s i s , pr e s e nc e of a s c i t e s )a nd wi t h r e c ur r e nc e - f r e e s ur v i v a l . The a i ms of our s t udy we r e t o c l a r i f y t he f unc t i ona l s i g ni f i c a nc e of CD1 4 7 i n c a nc e r pr og r e s s i on a nd t o e x a mi ne t he pos s i bi l i t y of us i ng CD1 4 7 a s a nov e l bi oma r ke r or a s a t he r a pe ut i c t a r g e t .
2 . Ge nomi c i de nt i f i c a t i on of s i g ni f i c a nt t a r g e t s i n c a nc e r a na l y s i s i n ov a r i a n c a r c i noma s We a na l y z e d s oma t i c DNA c opy numbe r v a r i a t i on i n v a r i ous hi s t ol og i c a l t y pe s of 7 8 ov a r i a n c a r c i noma s us i ng g e nomi c i de nt i f i c a t i on of s i g ni f i c a nt t a r g e t s i n c a nc e r a na l y s i s . Re g i ons t ha t we r e s i g ni f i c a nt l y( p- v a l ue ＜0 . 0 1 )mor e c ommon i n s e r ous t umor s ,a f t e r c or r e c t i ons f or mul t i pl e hy pot he s i s t e s t i ng( i ndi c a t i ng nonr a ndom di s t r i but i on a c r os s s ubt y pe s ) ,i nc l ude d( i n or de r of s i g ni f i c a nc e ) :8 p2 3 . 3 ,6 q2 4 . 3 ,1 1 p1 5 . 5 ,8 p2 1 . 2 ,1 6 q2 2 . 1 , 2 2 q1 3 . 3 1 ,4 q2 2 . 1 ,5 q2 2 . 2 ,7 p2 2 . 3 ,a nd 1 4 q2 4 . 2 . Al t houg h no r e g i ons of c opy numbe r v a r i a t i on we r e s i g ni f i c a nt l y a s s oc i a t e d wi t h a ny hi s t ol og i c a l s ubt y pe ot he r t ha n s e r ous t umor s ,a n ov e r r e pr e s e nt a t i on of a mpl i f i c a t i on of 2 0 q1 3 . 2( ZNF2 1 7 ) ,1 7 q1 2( CCL4 ) , a nd 8 q1 3 . 2 - q2 1 . 1 1( NCOA2 )wa s obs e r v e d i n c l e a r - c e l l t umor s ,a nd a mpl i f i c a t i on of 1 q2 1 . 3( CTMP)a nd 1 q4 2 . 1 3( RAB4 A)wa s obs e r v e d i n e ndome t r i oi d t umor s . 3 . J a pa ne s e ov a r i a n c a nc e r pa t i e nt s t ha t c a n c l a s s i f y t he pr e s e nc e of c a nc e r . The i nf or ma t i on i s s i mi l a r t o t ha t of s e r a of pa t i e nt s wi t h ov a r i a n c a nc e r i n t he Uni t e d St a t e s be c a us e N- di me ns i ona l c l us t e r s bui l t on Uni t e d St a t e s s e r a s pe c t r a c r e a t e d c l us t e r ma ps pr e di c t i v e of t he J a pa ne s e s a mpl e s . The pe r f or ma nc e of t he mode l wa s be t t e r t ha n t ha t
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of a ny of t he e x i s t i ng s i ng l e - bi oma r ke r a s s a y s , a l t houg h t r ul y us e f ul mode l s a wa i t a muc h l a r g e r s a mpl e s i z e a nd t he us e of i nde pe nde nt v a l i da t i on s a mpl e s e t s t o de mons t r a t e t he i r r obus t ne s s . The s e r e s ul t s e nc our a g e us t o s t a r t a l a r g e - s c a l e ,mul t i s i t e c ol l e c t i on of s e r a f r om J a pa ne s e pa t i e nt s wi t h ov a r i a n c a nc e r t o de v e l op a J a pa ne s e ov a r i a n c a nc e r s e r um pr of i l e a s s a y .
4 . Mi c r oRNA ( mi RNA)e x pr e s s i on pr of i l e s f or c a nc e r s ,i nc l udi ng t hos e of t he l ung , br e a s t ,s t oma c h,pr os t a t e ,a nd c ol on,we r e e x a mi ne d t o i nv e s t i g a t e t he i nv ol v e me nt of mi RNA i n c a r c i nog e ne s i s . We a r e now i nv e s t i g a t i ng t he r ol e s of mi RNA i n t he r e s i s t a nc e of huma n ov a r i a n c a nc e r c e l l s t o pa c l i t a x e l . Thi s r e s e a r c h ha s s i g ni f i c a nt i mpl i c a t i ons f or t he r a pe ut i c s t r a t e g i e s f or ov e r c omi ng c a nc e r c e l l c he mor e s i s t a nc e . 5 . Mi t oc hondr i a l ul t r a s t r uc t ur e - a s s oc i a t e d c he mot he r a py r e s pons e i n ov a r i a n c a nc e r A mi t oc hondr i a l s c or i ng s y s t e m wa s de v e l ope d i n a s s oc i a t i on wi t h pl a t i num s e ns i t i v i t y i n ov a r i a n c a nc e r a nd wa s a ppl i e d t o c l i ni c a l s a mpl e s . Twe nt y - f our c a s e s we r e i nc l ude d i n t hi s s t udy . Ex c e l l e nt c or r e l a t i on t o c he mos e ns i t i v i t y wa s not e d f or 2 f a c t or s :e l e c t r on de ns i t y a nd di s t r i but i on pa t t e r n. The t ot a l s c or e of t he s e 2 f a c t or s i n 9 s e ns i t i v e c a s e s wa s 1 . 4 4 a nd wa s 3 . 5 8 i n 1 9 r e s i s t a nt c a s e s( P ＜0 . 0 0 1 ) . Re c e pt or ope r a t i v e c ha r a c t e r i s - t i c s a na l y s i s r e v e a l e d t ha t t he t ot a l c ut - of f s c or e wa s 3 poi nt . Sc or e s wi t h t hi s s y s t e m we r e s t r ong l y c or r e l a t e d wi t h r e s pons e ,a nd t hi s r e s ul t s ug g e s t s t ha t t hi s s y s t e m woul d be of g r e a t v a l ue a s a bi oma r ke r f or c he mos e ns i t i v i t y .
6 . The pr og nos t i c f a c t or s i n pa t i e nt s wi t h a dv a nc e d e pi t he l i a l ov a r i a n c a nc e r ha v e be e n i de nt i f i e d by ma ny i nv e s t i g a t or s .We pe r f or me d a s t udy t o c ons t r uc t a s i mpl e a nd powe r f ul pr og nos t i c i nde x( PI )of e pi t he l i a l ov a r i a n c a nc e r( PI EPOC) . The f our pr og nos t i c f a c t or s t ha t r e ma i ne d i nde pe nde nt l y s i g ni f i c a nt i n t he a na l y s i s we r e a g e , pe r f or ma nc e s t a t us ,c e l l t y pe ,a nd s i z e of r e s i dua l di s e a s e . On t he ba s i s of t he r e g r e s s i on f unc t i on,pa t i e nt s we r e c l a s s i f i e d i nt o t hr e e r i s k g r oups ,i . e . ,l ow r i s k( PI :0 ‑ 2 ) ,i nt e r me - di a t e r i s k( PI :3 ) ,a nd hi g h r i s k( PI :4 ‑ 6 ) . The PI EPOC wa s e qua l l y pr e di c t i v e i n a v a l i da t i on s a mpl e( n ＝2 3 0 ) ,i de nt i f y i ng 3 g r oups( 5 - y e a r s ur v i v a l r a t e :6 7 % i n t he l ow- r i s k g r oup,4 3 %i n i nt e r me di a t e - r i s k g r oup,1 7 %i n t he hi g h- r i s k g r oup) .
7 . Se r um i ndol e a mi ne 2 , 3 - di ox y g e na s e e x pr e s s i on wa s pos i t i v e l y c or r e l a t e d wi t h i mpa i r e d s ur v i v a l i n pa t i e nt s wi t h s e r ous - t y pe ov a r i a n c a nc e r .
We ha v e pr e v i ous l y r e por t e d t ha t t he i ndol e a mi ne 2 , 3 - di ox y g e na s e( I DO)s c r e e ne d wi t h t he Ge ne Chi p i s pos i t i v e l y c or r e l a t e d wi t h pa c l i t a x e l r e s i s t a nc e a nd wi t h i mpa i r e d s ur v i v a l i n pa t i e nt s wi t h s e r ous - t y pe ov a r i a n c a nc e r . We e s t a bl i s he d a n e nz y me - l i nke d i mmunos or be nt a s s a y wi t h a n a nt i - I DO a nt i body f or s e r um a nd me a s ur e d s e r um I DO t i t e r s i n 2 6 t y pe s of ov a r i a n c a nc e r . We c ompa r e d t he e x pr e s s i on pa t t e r n i n s ur g i c a l s pe c i me ns a nd t he c or r e s pondi ng s e r um I DO t i t e r a nd f ound a pos i t i v e c or r e l a t i on i n t he s e r ous t y pe . The s e r e s ul t s s ug g e s t t ha t t he s e r um I DO l e v e l i s a bi oma r ke r f or s e r ous - t y pe ov a r i a n c a nc e r .
Fe t o mat e r nal me di c i ne
1 . Ant i phos phol i pi d s y ndr ome( APS)i s a c l i ni c a l e nt i t y ma ni f e s t e d by a r t e r i a l a nd v e nous t hr ombos e s a nd r e c ur r e nt mi s c a r r i a g e s ,a nd i s c a us e d by a nt i phos phol i pi d a nt i bodi e s .APS ha s a l s o be e n obs e r v e d i n s ome c ompl i c a t i ons of pr e g na nc y , e . g . , pr e g na nc y - i nduc e d hy pe r t e ns i on,i nt r a ut e r i ne g r owt h r e s t r i c t i on,a nd l a t e f e t a l de a t h.
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Howe v e r ,l i t t l e i s known a bout how a nt i phos phol i pi d a nt i bodi e s( APLs )i s i nv ol v e d i n t he s e c ompl i c a t i ons . To be i de nt i f i e d me c ha ni s ms of APLs i n obs t e t r i c a l c ompl i c a t i ons by i nv e s t i g a t i ng t he pa t hol og i c a l s i g ni f i c a nc e ,we e s t a bl i s he d a n e x pe r i me nt a l mode l f or APS us i ng wi l d t y pe ,Fc - knoc k- out mi c e a nd C- knoc kout mi c e .
2 . I DO pl a y s e s s e nt i a l r ol e s i n s uc c e s s f ul pr e g na nc y a nd i s i nduc e d by t y pe 1 c y t oki ne s . The r e c e pt or s of t y pe 1 c y t oki ne s s ha r e a s t r uc t ur e a nd a s i g na l t r a ns duc t i on pa t hwa y wi t h pr ol a c t i n. Be c a us e phy s i ol og i c a l l e v e l s of pr ol a c t i n i nc r e a s e ma r ke dl y dur i ng pr e g na nc y ,we e x a mi ne d a pos s i bl e c onne c t i on be t we e n pr ol a c t i n a nd I DO e x pr e s s i on.
To i nv e s t i g a t e I DO e x pr e s s i on i n CD1 4 c e l l s ,we obt a i ne d mononuc l e a r c e l l s f r om 1 2 he a l t hy pe r s ons a nd c ul t ur e d t he c e l l s f or 2 4 t o 4 8 hour s i n t he pr e s e nc e or a bs e nc e of s t i mul i( i nt e r f e r on ［I FN ］ - γ :5 , 1 0 0 I U / ml ;pr ol a c t i n:1 0 ,1 0 0 ,or 5 0 0 ng / ml ) . I DO e x pr e s s i on wa s up- r e g ul a t e d onl y whe n pr ol a c t i n a t phy s i ol og i c a l c onc e nt r a t i ons dur i ng pr e g na nc y we r e pr e s e nt wi t h 5 I U / ml of I FN- γ ,whe r e a s onl y pr ol a c t i n a nd 5 I U / ml of I FN- γ we r e wi t hout e f f e c t . The s e f i ndi ng s s ug g e s t pr ol a c t i n pl a y s i mpor t a nt r ol e s i n t he ma i nt e na nc e of pr e g na nc y ,t he r e by up- r e g ul a t i ng I DO,a nd s uppor t t he v i e w t ha t t he phy s i ol og i c a l c onc e nt r a t i on of I FN- γ i s i mpor t a nt f or pr e g na nc y .
3 . Es t a bl i s hme nt of a n i mmor t a l i z e d huma n e x t r a v i l l ous t r ophobl a s t c e l l l i ne by i nf e c t i on wi t h r e t r ov i r a l e x pr e s s i on v e c t or s
St udi e s of t he f unc t i on of huma n t r ophobl a s t s ha v e be e n l i mi t e d by a l a c k of s ui t a bl e c e l l mode l s . We a i me d t o obt a i n huma n nor ma l t r ophobl a s t c e l l l i ne s wi t h l ong l i f e s pa ns a nd c ons e que nt l y pr ov i de a n i de a l i n v i t r o c e l l mode l . Pr i ma r y huma n t r ophobl a s t c e l l s we r e de r i v e d f r om t he pl a c e nt a of a woma n unde r g oi ng e l e c t i v e a bor t i on dur i ng t he 7 t h we e k of g e s t a t i on. The c e l l s we r e i mmor t a l i z e d by i nf e c t i on wi t h r e t r ov i r a l e x pr e s s i on v e c t or s c ont a i ni ng t y pe 1 6 huma n pa pi l l oma v i r us E6 a nd E7 i n c ombi na t i on wi t h huma n t e l ome r a s e r e v e r s e t r a ns c r i pt a s e . The c ha r a c t e r i s t i c s of t he c e l l l i ne we r e a na l y z e d. I mmunoc y t oc he mi c a l s t udi e s r e v e a l e d s t a i ni ng f or huma n c hor i oni c g ona dot r ophi n c ha i nβ ,c y t oke r a t i n 7 ,HLA- G,a nd CD9 a nd a n e x t r a v i l l ous t r ophobl a s t i c phe not y pe . Tr a ns we l l i ns e r t i nv a s i on a s s a y s howe d t he i nv a s i v e ne s s of t hi s c e l l l i ne ,a nd g e l a t i n z y mog r a phy de t e c t e d t he s e c r e t i on of MMP- 2 a nd MMP- 9 . Ka r y ot y pe a na l y s i s s howe d ne a r l y nor ma l c hr omos oma l numbe r s wi t h s ma l l de v i a t i ons r a ng i ng f r om 4 6 t o 4 8 ,a nd nude mous e a s s a y s howe d no t umor i g e ne c i t y . Thi s ne wl y i mmor t a l i z e d c e l l l i ne ,HChEpC1 b,wi l l pr ov i de a us e f ul mode l f or t he s t udy of e x t r a v i l l ous t r ophobl a s t f unc t i on.
Re p r o duc t i v e e ndo c r i no l o g y
1 . Mos t of t he me c ha ni s ms f or a c hi e v i ng pr e g na nc y ha v e be e n c l a r i f i e d due t o t he de v e l opme nt of a s s i s t e d r e pr oduc t i v e t e c hnol ol g i e s . Howe v e r ,t he me c ha ni s m of i m- pl a nt a t i on r e ma i ns unc l e a r .
CD1 4 7 i s e x pr e s s e d a t hi g h l e v e l s on t he s ur f a c e s of v a r i ous t umor c e l l s a nd s t i mul a t e s MMPs . We hy pot he s i z e d t ha t CD1 4 7 pl a y s a n i mpor t a nt r ol e i n i mpl a nt a t i on. The a i m of t hi s s t udy wa s t o de t e r mi ne t he e x pr e s s i on a nd hor mona l r e g ul a t i on of t he CD1 4 7 g e ne dur i ng t he huma n i mpl a nt a t i on pe r i od i n c ont r ol l e d ov a r i a n hy pe r s t i mul a t i on c y c l e s . We f ound t ha t CD1 4 7 a nd MMP- 2 mRNAs i n huma n e ndome t r i um dur i ng t he
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s e c r e t or y pha s e s we r e s i g ni f i c a nt l y de c r e a s e d i n c ont r ol l e d ov a r i a n hy pe r s t i mul a t i on c y c l e s .
Publications
Isonishi S,Saitou M,Sai tou M,Yasuda M,Tanaka T.Differential regulation of the cytotoxicity activity of paclitaxel by orobol and platelet der- ived growth factor in human ovarian carcinoma cells.Oncol Rep 2007;18:195‑201.
Teramukai S,Ochiai K,Tada H,Fukushima M.
PIEPOC:a new prognostic index for advanced epithelial ovarian cancer-Japan Multinational Trial Organization OC01‑01.J Clon Oncol 2007;25:3302‑6.
Saeki T,Sano M,Komoike Y,Sonoo H,Honjyo H, Ochiai K,Kobayashi T,Aogi K,Sato N,Sawai S, Miyoshi Y,Takeuchi M,Takashima S.No increase of breast cancer incidence in Japanese women who received hormone replacement therapy:overview of a case-control study of breast cancer risk in Japan.Int J Clin Oncol 2008;13:8‑11.
Motegi M,Takakura S,Takano H,Tanaka T, Ochiai K.Adjuvant chemotherapy in a pregnant patient with endometrial sinus tumor of the ovary: a case report.Obstet Gynecol 2007;109:537‑ 40.
Itoh H,Iwasaki M,Hanaoka T,Sasaki H,Tanaka T,Tsugane S.Urinary bisphenol-A concentra- tion in infertile Japanese women and its associa- tion with endometriosis:a cross-sectional study. Environ Health Prev Med 2007;12:258‑64.
Isonishi S,Coleman RL,Hirama M,Iida Y,Kitai S, Nagase M,Ochiai K.Analysis of prognostic factors for patients with leiomyoma treated with uterine arterial embolization.Am J Obstet Gynecol 2008;198:270.
Ueda K,Yamada K,Urashima M,Ishibashi Y, Shirai M,Nikaido T,Takahashi H,Okamoto A,
Saito M,Yasuda M,Ohkawa K,Tanaka T.
Association of extracellular matrix metallo- proteinase inducer in endometrial carcinoma with patient outcomes and clinicopathologenesis using monoclonal antibody 1 2C3.Oncol Rep 2007;17:731‑5.
Takao M,Okamoto A,Nikaido T,Urashima M, Takakura S,Saito M,Saito M,Okamoto S, Takikawa O,Sasaki H,Yasuda M,Ochiai K, Tanaka T.Increased synthesis of indoleamine- 2,3-dioxygenase protein is positively associated with impaired survival in patients with serous- type,but not with other types of,ovarian cancer. Oncol Rep 2007;17:1333‑9.
Takakura S,Saito M,Ueda K,Motegi M,Takao M, Yamada K,Okamoto A,Niimi S,Sasaki H, Tanaka T,Ociai K.Irinotecan hydrochloride (CPT-11)and cisplatin as first-line chemotherapy after surgery for ovarian clear cell adenocar- cinoma.Int Surg 2007;92:202‑8.
Seike M,Yanaihara N,Bowman ED,Zanetti KA, Budhu A,Kumamoto K,Mechani c LE,Matsumoto S,Yokota J,Shibata T,Sugimura H,Gemma A, Kudoh S,Wang XW,Harris CC.Use of a cyto- kine gene expression signature in lung adenocar- cinoma and the surrounding tissue as a prognos- tic classifier.J Natl Cancer Inst 2007;99:
1257‑69.
Kawaguchi R, Shimokawa T, Umehara N, Nunomura S,Tanaka T,Ra C.Priming of peripheral monocytes with prolactin(PRL)sensi- tizes IFN-γ-mediated indoleamine 2,3- dioxygenase(IDO)expr ession without affecting IFN-γsignaling.J Reprod Immunol 2008;7:
117‑25.
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