Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
권호 표지

Comparison of Gene Expression Profile in Eutopic Endometria with or without Endometriosis: A Microarray Study

자궁내막증 환자와 대조군에서의 자궁내막 유전자 발현의 차이: Microarray를 이용한 연구

  • Chung, Min-Ji (Department of Obstetrics and Gynecology, Department of Immunology, Kyungpook National University, School of Medicine) ;
  • Chung, Eun-Jung (Department of Obstetrics and Gynecology, Kyungpook National University, School of Medicine) ;
  • Lee, Shin-Je (Department of Obstetrics and Gynecology, Kyungpook National University, School of Medicine) ;
  • Kim, Moon-Kyu (Department of Obstetrics and Gynecology, Kyungpook National University, School of Medicine) ;
  • Chun, Sang-Sik (Department of Obstetrics and Gynecology, Department of Immunology, Kyungpook National University, School of Medicine) ;
  • Lee, Taek-Hoo (Department of Obstetrics and Gynecology, Department of Immunology, Kyungpook National University, School of Medicine)
  • 정민지 (경북대학교 의과대학 산부인과학교실) ;
  • 정은정 (경북대학교 의과대학 면역학교실) ;
  • 이신제 (경북대학교 의과대학 면역학교실) ;
  • 김문규 (경북대학교 의과대학 면역학교실) ;
  • 전상식 (경북대학교 의과대학 산부인과학교실) ;
  • 이택후 (경북대학교 의과대학 산부인과학교실)
  • Published : 2007.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Objective: Pathogenesis of the endometriosis is very complex and the etiology is still unclear. Our hypothesis is that there may be some difference in gene expression patterns between eutopic endometriums with or without endometriosis. In this study, we analyzed the difference of gene expression profile with cDNA microarray. Methods: Endometrial tissues were gathered from patients with endometriosis or other benign gynecologic diseases. cDNA microarray technique was applied to screen the different gene expression profiles from early and late secretory phase endometria of those two groups. Each three mRNA samples isolated from early and late secretory phase of endometrial tissues of control were pooled and used as master controls and labeled with Cy3-dUTP. Then the differences of gene expression pattern were screened by comparing eutopic endometria with endometriosis, which were labeled with Cy5-dUTP. Fluorescent labeled probes were hybridized on a microarray of 4,800 human genes. Results: Twelve genes were consistently over-expressed in the endometrium of endometriosis such as ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ transporting (ATP5C1) and TNF alpha factor. Eleven genes were consistently down-regulated in the endometriosis samples. Many extracellular matrix protein genes (decorin, lumican, EGF-containing fibulin-like extracellular matrix protein 1, fibulin 5, and matrix Gla protein) and protease/protease inhibitors (serine proteinase inhibitor, matrix metalloproteinase 2, tissue inhibitor of metalloproteinase 1), and insulin like growth factor II associated protein were included. Expression patterns of selected eight genes from the cDNA microarray were confirmed by quantitative RT-PCR or real time RT-PCR. Conclusion: The result of this analysis supports the hypothesis that the endometrium from patients with endometriosis has distinct gene expression profile from control endometrium without endometriosis.

목 적: 자궁내막증은 자궁내부에 존재하여야 할 자금내막조직이 자궁 외에 존재하는 질환으로 그 발생기전은 아직 명확하게 밝혀져 있지 않다. 이에 저자들은 자궁내막증 환자와 정상 대조군의 자궁내막조직 간의 유전자 발현의 차이가 자궁내막증의 발병과 관련이 있을 것이라는 가정 하에 DNA microarray 기술을 도입하여 연구를 시행하였다. 연구방법: 2002년 1월부터 2002년 12월까지의 기간 동안 본원 산부인과에서 자궁내막증 환자와 자궁내막증 이외의 다른 부인과적 질환으로 수술을 시행한 환자들을 대상으로 채취한 자궁내막 조직으로 KNU 4.8K cDNA chip을 이용하여 유전자 발현을 비교 연구하였다. 유전자칩으로 자궁내막증 조직에서 발현의 증감을 보였던 유전자 중에서 8종의 유전자를 대상으르 RT-PCR이나 real time RT-PCR 법을 통하여 그 발현 양상을 검증하였다. 결 과: 자궁내막증에 이환된 여성의 자궁내막조직에서 대조군에 비하여 높게 발현되고 있는 것으로 나타난 유전자들은 ATP synthase H transporting F1 (ATP5B), eukaryotic translation elongation factor 1, isocitrate dehydrogenase 1 (NADP+), mitochondrial ribosomal protein L3, ATP synthase H+ trarsporting (ATP5C1), LPS induced TNF-$\alpha$ factor 등으로 세포의 에너지 생성과 대사과정 및 신호전달에 관여하는 유전자들이었다. 한편 자궁내막중 환자의 자궁내막조직에서 대조군에 비하여 낮게 발현된 유전자들은 insulin like growth factor II associated protein, EGF-containing fibulin-like EMP1, matrix Gla protein, TGF beta-induced, TGF beta receptor 1(activin A receptor type II-like kinase), cystallin alpha B, fibulin 5, tissue inhibitor of metalloproteinase 3, collage type XII, alpha 1, tissue inhibitor of metalloproteinase 1, decorin 등으로 세포외기질의 구성 및 기능에 관련이 있었다. 결 론: 이상의 DNA mirroarry 및 RT-PCR을 통해 얻어진 결과에서 자궁내막증의 자궁내막조직에서 대조군에 비하여 유전자들의 발현에 차이가 있음을 확인하였다.

Keywords

References

  1. Sampson JA. Peritoneal endometriosis due to the menstrual dissemination of endometrial tissue into the peritoneal cavity. Am J Obstet Gynecol 1927; 14: 422
  2. Berbieri RL. Etiology and epidemiology of endometriosis. Am J Obstet Gynecol 1990; 162: 565-7 https://doi.org/10.1016/0002-9378(90)90430-F
  3. Mahmood TA, Templeton A. Folliculogenesis and ovulation in infertile women with mild endometriosis. Hum Reprod 1991; 6: 227-31 https://doi.org/10.1093/oxfordjournals.humrep.a137311
  4. Halme J, Hammond MG, Hulka JF, Raj SG, Talbert LM. Retrograde menstruation in healthy women and in patients with endometriosis. Obstet Gynecol 1984; 64: 151-4
  5. Lamb K, Hoffman RG, Nichols TR. Family trait analysis: A case-control study of 43 women with endometriosis and their best friends. Am J Obstet Gynecol 1986; 154: 596
  6. Cramer DW, Wilson E, Stillman RJ, Berger MJ, Belisle S, Schiff I, et al. The relation of endometriosis to menstrual characteristics, smoking, and exercise. JAMA 1986; 255: 1904-8 https://doi.org/10.1001/jama.255.14.1904
  7. Ramey JW, Archer DE Peritoneal fluid: its relevance to the development of endometriosis. Fertil Steril 1993; 60: 1 https://doi.org/10.1016/S0015-0282(16)56027-0
  8. Oosterlynck DJ, Comillie FJ, Waer M, Vandeputte M, Koninckx PR. Women with endometriosis show a defect in natural killer activity resulting in a decreased cytotoxicity to autologous endometrium. Fertil Steril 1991; 56: 45-51 https://doi.org/10.1016/S0015-0282(16)54414-8
  9. Claverie JM. Computational methods for the identification of differential and coordinated gene expression. Hum Mol Genet 1999; 8: 182133 https://doi.org/10.1093/hmg/8.12.2133
  10. Levenstien MA, Yang Y, Ott J. Statistical significance for hierarchical clustering in genetic association and microarray expression studies. BMC Bioinformatics 2003; 4: 62-70 https://doi.org/10.1186/1471-2105-4-62
  11. Badawy SZA, Cuenca V, Marshall L, et al. Cellular components in peritoneal fluid in infertile patterns with and without endometriosis. Fertil Steril 1984; 42: 704 https://doi.org/10.1016/S0015-0282(16)48194-X
  12. Bartosik D, Jacobs SL, Kelly LJ. Endometrial tissue in peritoneal fluid. Fertil Steril 1986; 46: 796 https://doi.org/10.1016/S0015-0282(16)49813-4
  13. Gompel C, Silverberg SG. The female peritoneum. Pathology in gynecology and obstetrics, 3rd edn., Philadelphia, U.S.A., JB Lippincott 1985; 411
  14. Steck WD, HElwing EB. Cutaneous endometriosis. Clin Obstet Gynecol 1966; 9: 373 https://doi.org/10.1097/00003081-196606000-00007
  15. Halban J. Hysteroadenosis rnetastatica: die lymphogene genese der so. Adenofibromatosis heterotropica. Wien KIin Woochenschr schr 1924; 37: 1205
  16. Simpson JL, Elias S, Malinak LR, Buttram VCJ. Heritable aspects of endometriosis. I Genetic studies. Am J Obstet Gynecol 1980; 137: 327-31 https://doi.org/10.1016/0002-9378(80)90917-5
  17. Coxhead D, Thomas EJ. Familial inheritance of endometriosis in a British population. A case control study. J Obstet Gynecol 1993; 13: 42-4 https://doi.org/10.3109/01443619309151773
  18. Witz CA. Interleukin-6: another piece of the endometriosis-cytokine puzzle. Fertil Steril 2000; 73: 212-4 https://doi.org/10.1016/S0015-0282(99)00556-7
  19. Koninckx PR, Barlow D, Kennedy S. Implantation versus infiltration: the Sampson versus the endometriotic disease theory. Gynecol Obstet Invest 1999; 47: 3-10 https://doi.org/10.1159/000052853
  20. Iwabe T, Harada T, Tsudo T, Tanikawa M, Oonahara Y, Terakawa N. Pathogenetic significance of increased levels of interleukin-8 in the peritoneal fluid of patients with endometriosis. Fertil Steril 1998; 69: 924-30 https://doi.org/10.1016/S0015-0282(98)00049-1
  21. Iba Y, Harada T, Horie S, Deura I, Iwabe T, Terakawa N. Lippopolysaccharide-promoted proliferation of endometriotic stromal cells via induction of tumor necrosis factor a and interleukin-8 expression. Fertil Steril 2004; 82: 1036-42 https://doi.org/10.1016/j.fertnstert.2004.04.038
  22. Matsuda A, Suzuki Y, Honda G, Muramatsu S, Matsuzaki O, Nagano Y, et al. Large-scale identification and characterization of human genes that activate NF-$\kappa$B and MAPK signaling pathways. Oncogene 2003 May 22; 22(21): 3307-18
  23. Mckay LI, Cidlowski JA. Molecular control of inunune/inflammatory responses; interactions between nuclear factor-kappa B and steroid receptorsignaling pathways. Endoc Rev 1999; 20: 435-59 https://doi.org/10.1210/er.20.4.435
  24. Yamauchi N, Harada T, Taniguchi F, Yoshida S, Iwabe T, Terakawa N. Tumor necrosis factor-$\alpha$ induced the release of interleukin-6 from endometriotic stromal cells by the nuclear factor-$\kappa$B and mitogen-activated protein kinase pathways. Fertil Steril 2004; 82: 1023-8 https://doi.org/10.1016/j.fertnstert.2004.02.134
  25. Goldman R. Growth factors and chronic wound healing: past, present, and future. Adv Skin Wound Car. 2004; 17: 24 https://doi.org/10.1097/00129334-200401000-00012
  26. Irwin JC, De las Fuentes L, Dsupin BA, Giudice LC. Insulin-like growth factor regulation of human endometrial stromal cell function: coordinate effects on IGF binding protein-1, cellular proliferation, and prolactin secretion. Regul Peptides 1993; 48: 165-77 https://doi.org/10.1016/0167-0115(93)90345-9
  27. Sbracia M, Zupi E, Alo P, Manna C, Marconi D, Scarpellini F, et al. Differential expression of IGF-I and IGF-II in eutopic and ectopic endometria of women with endometriosis and in women without endometriosis. Am J Reprod Immunol 1997; 37(4): 326-9 https://doi.org/10.1111/j.1600-0897.1997.tb00238.x