체외 배양된 자궁내막세포에서의 DOC-1 유전자의 발현 조절

In Vitro Regulation of DOC-1 Gene Expression in Uterine Endometrial Cells

  • 양혜영 (성신여자대학교 자연과학대학 기초과학연구소, 생물학과) ;
  • 전용필 (성신여자대학교 자연과학대학 기초과학연구소, 생물학과)
  • Yang, Hye-Young (Division of Developmental Biology and Physiology, School of Biological Sciences and Chemistry, Institute of Basic Sciences, Sungshin Women's University) ;
  • Cheon, Yong-Pil (Division of Developmental Biology and Physiology, School of Biological Sciences and Chemistry, Institute of Basic Sciences, Sungshin Women's University)
  • 발행 : 2009.12.31

초록

포배의 착상은 구조 및 기능적으로 준비된 자궁내막의 체계적 반응에 의하여 진행되는데, 자궁 내막에서 엄격히 통제된 급격한 세포의 증식과 분화가 진행된다. 구강 상피세포암의 억제자로 알려진 Doc-1이 자궁에서 스테로이드 호르몬에 의하여 발현되며 세포 증식을 조절할 것으로 추정되어 왔으나, 그 정확한 발현 변화에 대한 이해가 불충분하였다. 따라서 본 연구에서는 체외에서 탈락막 반응 모델 확립을 통하여 세포의 증식과 분화 진행과정에서 Doc-1의 발현 조절기작을 알아보고자 하였다. 생쥐 자궁내막을 이용한 탈락막 반응은 기질세포를 순수하게 분리하여 배양하면서 프로게스테론과 에스트로겐을 함유한 유도 배양액으로 유도하였다. 탈락막 유도 과정에서 배양 24시간까지 세포의 증식이 유의하게 증가하였고 그 후에는 감소하였다. 한편, 탈락막 세포로의 분화는 분화 유도 48시간에 거의 모든 세포에서 진행되었다. 또한 Doc-1 단백질의 발현이 분화 유도 시간과 비례적으로 증가하였고 탈락막 세포에 위치하였다. 이러한 결과를 바탕으로 착상이 진행되는 자궁내막에서 Doc-1의 발현이 스테로이드 호르몬과 탈락막 분화와 연계되어 조절되고, Doc-1 단백질이 탈락막 세포의 세포분열 억제를 유도하는 것을 제안할 수 있다.

Implantation of blastocyst into the uterine endometrium is established by the existence of histologically and functionally prepared uterine endometrium. Doc-1, an oral cancer suppressor gene, is expressed under the control of steroid hormones and has been suggested as a proliferation regulator of endometrial cells. However, the role is not much clear and in this study we examined the expression modulation of Doc-1 in decidualizing cells in vitro. In vitro decidualization was performed in endometrial stroma cells using progesterone and estrogen. Until 24 hr after decidual induction the proliferation of stroma cell was significantly increased but decreased after then. On the other hand, most of the cells differentiated into decidual cell after 48 hr of induction. The Doc-1 protein was co-localized in a specific deciudal cells and colocalization rate was increased in a parallel manner with the induction time. Based on these results, it is suggested that Doc-1 expression is under the control of both steroid hormones and decidual signals, and Doc-1 protein is involved in suppression of the proliferation of decidualizing cells.

키워드

참고문헌

  1. Bijovsky AT, Zorn TM, Abrahamsohn PA (1992) Remodeling of the mouse endometrial stroma during the preimplantation period. Acta Anat 144:231-234. https://doi.org/10.1159/000147311
  2. Brar AK, Frank GR, Kessler CA, Cedars MI, Handwerger S (1997) Progesterone-dependent decidualization of the human endometrium is mediated by cAMP. Endocrine 6:301-307. https://doi.org/10.1007/BF02820507
  3. Cheon YP (2002) Expression of Doc-1 in pregnant uterus of the mouse. Kor J Fetil Steril 29:295-302.
  4. Cheon YP, Kim CH (In press) Progesterone is Primary Regulator of Cdk2ap1 gene Expression and Tissuespecific Expression in the Uterus. J Endocrinol Invest
  5. Cheon YP, Li Q, Xu X, DeMayo FJ, Bagchi I, Bagchi MK (2002) A genomic approach to identify novel progesterone receptor regulated pathways in the uterus during implantation. Mol Endocrinol 16:2853-2871. https://doi.org/10.1210/me.2002-0270
  6. Dey SK, Lim H, Das SK, Reese J, Paria BC, Daikoku T, Wang H (2004) Molecular cues to implantation. Endocrine Rev 25:341-373. https://doi.org/10.1210/er.2003-0020
  7. Geum D, SunW, Paik SK, Lee CC, Kim K (1997) Estrogeninduced cyclin D1 and D3 gene expressions during mouse uterine cell proliferation in vivo: differential induction mechanism of cyclin D1 and D3. Mol Reprod Dev 46:450-458. https://doi.org/10.1002/(SICI)1098-2795(199704)46:4<450::AID-MRD2>3.0.CO;2-N
  8. Goffin F, Munaut C, Malassine A, Evain-Brion D, Frankenne F, Fridman V, Dubois M, Uzan S, Merviel P, Foidart JM (2003) Evidence of a limited contribution of fetomaternal interactions to trophoblast differentiation along the invasive pathway. Tissue Antigens 62:104-116. https://doi.org/10.1034/j.1399-0039.2003.00085.x
  9. Gordon HM, Kucera G, Salvo R, Boss JM (1992) Tumor necrosis factor induces genes involved in inflammation, cellular and tissue repair, and metabolism in murine fibroblasts. J Immunol 148:4021-4027.
  10. Imakawa K, Chang KT, Christenson RK (2004) Pre-implantation conceptus and maternal uterine communications: molecular events leading to successful implantation. J Reprod Dev 50:155-169. https://doi.org/10.1262/jrd.50.155
  11. Kim H, Cheon YP (2006). Spatio-temporal expression and regulation of dermatopontin in the early pregnant mouse uterus. Mol Cells 31:262-268.
  12. Kimura F, Takakura K, Takebayashi K, Ishikawa H, Kasahara K, Goto S, Noda Y (2001) Messenger ribonucleic acid for the mouse decidual prolactin is present and induced during in vitro decidualization of endometrial stromal cells. Gynecol Endocrinol 15:426-432.
  13. Ledee N, Dubanchet S, Oger P, Meynant C, Lombroso R, Ville Y, Chaouat G (2007) Uterine receptivity and cytokines: new concepts and new applications. Gynecol Obstet Invest 64:138-143. https://doi.org/10.1159/000101737
  14. Lee S, Lee SA, Shim CS, Khang IK, Lee K-A, Park Y-M, Kang B-M, Kim KJ (2003) Identification of estrogenregulated genes in the mouse uterus using a delayedimplantation model. Mol Reprod Dev 64:405-413. https://doi.org/10.1002/mrd.10232
  15. Mantena SR, Kannan A, Cheon YP, Li Q, Jonson PF, Bagchi, IC, Bagchi MK (2006) C/EBPbeta is a critical mediator of steroid hormone-regulated cell proliferation and differentiation in the uterine epithelium and stroma. Proc Natl Acad Sci USA 103:1870-1875. https://doi.org/10.1073/pnas.0507261103
  16. Mulholland J, Aplin JD, Ayad S, Hong L, Glasser SR (1992) Loss of collagen type VI from rat endometrial stroma during decidualization. Biol Reprod 46:1136- 1143. https://doi.org/10.1095/biolreprod46.6.1136
  17. Orlando-Mathur CE, Bechberger JF, Godberg GS, Nus CC, Kidder GM, Kennedy TG (1996) Rat endometrial stromal cells express the gap junction genes connexins 26 and 43 and form functional gap junctions during in vitro decidualization. Biol Reprod 54:905-913. https://doi.org/10.1095/biolreprod54.4.905
  18. Prall OWJ, Sarcevic B, Musgrove EA, Watts CKW, Sutherland RL (1997) Estrogen-induced activation of cdk4 and cdk2 during G1-S phase progression is accompanied by increased cyclin D1 expression and decreased cyclindependent kinase inhibitor association with cyclin E-cdk2. J Biol Chem 272:10882-10894. https://doi.org/10.1074/jbc.272.16.10882
  19. Shimizu A, Maruyama T, Tamaki K, Uchida H, Asada H, Yoshimura Y (2005) Impairment of decidualization in SRC-deficient mice. Biol Reprod 73:1219-1227. https://doi.org/10.1095/biolreprod.105.041616
  20. Shintani S, Ohyama H, Zhang X, McBride J, Matsuo K, Tsuji T, Hu MG, Hu G, Kohno Y, Lerman M, Todd R, Wong DT (2000) p12(DOC-1) is a novel cyclin dependent kinase 2-associated protein. Mol Cell Biol 20:6300-6307. https://doi.org/10.1128/MCB.20.17.6300-6307.2000
  21. Shiozawa T, Li SF, Nakayama K, Nikaido T, Fujii S (1996) Relationship between the expression of cyclins/ cyclin-dependent kinases and sex-steroid receptors/Ki67 in normal human endometrial glands and stroma during the menstrual cycle. Mol Hum Reprod 2:745-752. https://doi.org/10.1093/molehr/2.10.745
  22. Tang B, Gurpide E (1993) Direct effect of gonadotropins on decidualization of human endometrial stromal cells. J Steroid Biochem Mol Biol 47:115-121. https://doi.org/10.1016/0960-0760(93)90064-4
  23. Todd R, McBridge J, Tsuji T, Donoff RB, Nagai M, Chou MY, Chiang T, Wong DT (1995) Deleted in oral cancer-1 (doc-1), a novel oral tumor suppressor gene. FASEB J 9:1362-1370.
  24. Tong W, Pollard JW (1999) Progesterone inhibits estrogeninduced cyclin D1 and cdk4 nuclear translocation, cyclin E- and cyclin A-cdk2 kinase activation, and cell proliferation in uterine epithelial cells in mice. Mol Cell Biol 19:2251-2264.
  25. Tsuji T, Duh FM, Latif F, Popescu NC, Zimonjic DB, McBride J, Matsuo K, Ohyama H, Todd R, Nagata E, Terakado N, Sasaki A, matsumura T, Lerman MI, Wong DT (1998) Cloning, mapping, expression, function, and mutation analyses of the human ortholog of the hamster putative tumor suppressor gene Doc-1. J Biol Chem 273:6704-6709. https://doi.org/10.1074/jbc.273.12.6704
  26. Weitlauf HM (1994) Biology of implantation. In: Knobil E, Neil JD (ed.), The Physiology of Reproduction. Raven, New York, pp391-448.