Tesis

Hipermetilasi Promotor Gen dan Ekspresi mRNA Reseptor Progesteron A dan B pada Jaringan Endometrioma = Gene Promoter Hypermethylation and mRNA Expression of A and B Progesterone Receptor in Endometrioma.

LATAR BELAKANG: Endometriosis merupakan penyakit ginekologis kronis yang ditandai dengan adanya jaringan mirip endometrium diluar rongga uterus. Endometriosis merupakan penyebab infertilitas dan nyeri pelvik paling umum dan memengaruhi 1 dari 10 wanita usia reproduktif. Progesteron memiliki peran yang penting dalam uterus yaitu mengontrol proliferasi dan diferensiasi. Disregulasi progesteron dapat menyebabkan gangguan fungsi uterus. Resistensi progesteron banyak ditemukan pada endometriosis dan dikaitkan dengan kadar PGR yang rendah. PGR memiliki dua isoform yaitu PGR-A dan PGR-B. Hingga saat ini masih banyak pendapat yang berbeda mengenai ekspresi isoform PGR-A dan B pada endometriosis. Hipermetilasi pada daerah promotor suatu gen diyakini sebagai salah satu penyebab gene silencing yang berakibat rendahnya kadar gen tersebut. Penelitian ini bertujuan untuk mengetahui pengaruh faktor epigenetik pada reseptor hormon progesteron pada jaringan endometriosis. METODE: Penelitian ini merupakan studi kasus kontrol yang membandingkan 20 wanita penderita endometriosis dan 20 wanita bukan penderita endometriosis. Analisis metilasi DNA dilakukan dengan metode MSP dan ekspresi mRNA dengan metode qPCR. Analisis statistik yang dilakukan adalah uji t tidak berpasangan, uji Mann Whitney, uji korelasi Pearson, uji korelasi Spearman’s Rho, Uji Annova One Way dan Uji Kruskal-wallis dengan kemaknaan p0,05). KESIMPULAN: Daerah promotor gen PGR mengalami hipermetilasi pada endometriosis dibandingkan dengan kontrol dan berkaitan dengan penurunan kadar PGR-B pada endometriosis.
Kata Kunci: metilasi DNA, ekspresi mRNA, PGR, endometriosis



Background: Endometriosis is a chronic gynecological disorder, defined as the presence of endometrium-like tissue outside of the uterine cavity. Endometriosis is the most common causes of infertility and pelvic pain and affects 1 of 10 women in the reproductive-age group. Progesterone plays an important role in uterine. It controls endometrial proliferation and differentiation. Dysregulation of progesterone signaling leads to impaired for uterine function. Progesterone resistance has been found in endometriosis and associated with the low levels of PGR. PGR has two isoforms, PGR-A and PGR-B. Since promoter hypermethylation is associated with gene silencing, we try to determine the methylation status of PGR promoter region in endometriosis tissue using methylation spesific PCR and its association with the expression of PGR-A and PGR-B isoforms using real-time PCR. Methods: this research is a case-control study, comparing 20 women with endometriosis and 20 women without endometriosis. Methylation status was analyzed with methylaion spesific PCR and the expression of mRNA was analyzed with real-time PCR. Statistical analyses were t-independent test, Mann Whitney test, Pearson test, Spearman’s rho test, Annova One Way test and Kruskal-Wallis test, a two-tailed p value less than 0,05 was considered significant. Result: We found that methylation status in women with endometriosis (98,72%) was significantly higher than women without endometriosis (3,74%), statistically significant associations with the disease (p=0,000). Beside, mRNA expression of PGR-A and PGR-B isoform was down regulated (2,35 fold and 6,37 fold). Correlation between methylation status and PGR-B expression was significant (p=0,000;r=-0,736), but not PGR-A (p>0,05). Conclusion: Promoter regions of PGR is hypermethylated in endometriosis as compared with control. This findings suggest that the promoter hypermethylation of PGR may contribute to the pathogenesis of this disease.
Keywords: DNA methylation, mRNA expression, PGR, endometriosis