Skripsi

Analisis Metilasi DNA pada Gen Plasminogen Activator Inhibitor-1 di Jaringan Endometriosis Peritoneum dan Ovarium Pasien Penderita Endometriosis = Methylation Analysis of Plasminogen Activator Inhibitor-1 Gene in Ovarian and Peritoneal Endometriosis.

Endometriosis adalah penyakit ginekologis multifaktorial kronis yang ditandai dengan adanya pertumbuhan jaringan mirip endometrium di luar rahim. Jaringan ini memiliki kemampuan untuk tertanam di ovarium, peritoneum, dan lokasi ektopik lainnya. Kemampuan untuk tertanam dipengaruhi oleh sistem aktivator plasminogen yang diketahui berperan dalam proses fibrinolisis. Kurangnya fibrinolisis akibat berlebihnya ekspresi plasminogen activator inhibitor-1 (PAI-1) telah ditemukan terjadi pada jaringan endometriosis. Kegagalan fibrinolisis yang terjadi menyebabkan terbentuknya produk fibrin terdegradasi yang dapat menempel dan berkembang menjadi endometriosis. Telah diketahui bahwa faktor epigenetik berperan pada patogenesis endometriosis, salah satunya adalah perubahan pada tingkat metilasi DNA. Penelitian ini bertujuan untuk menilai tingkat metilasi gen PAI-1 dan hubungannya dengan perkembangan jaringan endometriosis ovarium dan peritoneum. Studi potong lintang ini menggunakan sampel dari 13 wanita dengan endometriosis ovarium, 11 wanita dengan endometriosis peritoneum, dan 11 wanita tanpa endometriosis. DNA dari sampel diisolasi, dilakukan konversi bisulfit, dan kemudian pengamatan tingkat metilasi DNA dilakukan dengan metode methylation specific polymerase chain reaction (MSP) yang diikuti oleh proses elektroforesis. Hasilnya dianalisis secara statistik dengan uji Kruskal-Wallis yang dilanjutkan dengan uji Mann-Whitney. Terdapat perbedaan yang signifikan secara statistik tingkat metilasi DNA gen PAI-1 pada endometriosis ovarium, endometriosis peritoneum, dan kontrol (p=0,000). Penelitian ini juga menemukan perbedaan signifikan antara endometriosis ovarium dan peritoneum masing-masing dibandingkan dengan kontrol (p=0,001 dan p = 0,000); namun tidak ada perbedaan tingkat metilasi DNA gen PAI-1 yang signifikan pada endometriosis peritoneum dibandingkan dengan endometriosis ovarium (p=0,111). Penelitian kami menunjukkan rendahnya tingkat metilasi gen PAI-1 yang dapat meningkatkan ekspresi gen PAI-1 dan hal ini disugestikan dapat berkontribusi sebagai faktor risiko endometriosis pada ovarium dan peritoneum.
Kata kunci: Endometriosis ovarium, Endometriosis peritoneum, Gen PAI-1, Metilasi DNA


Endometriosis is a chronic multifactorial gynecological disease characterized by the growth of endometrial-like tissue outside the uterus. This tissue can be embedded in the ovaries, peritoneum, and other ectopic locations. The ability is influenced by a plasminogen activator system which is known to play a role in the fibrinolysis process. The defect of fibrinolysis due to excess expression of plasminogen activator inhibitor-1 (PAI-1) has been found to occur in endometriosis tissue. Fibrinolysis defect that occurs causes the formation of fibrin degradation product that can attach and develop into endometriosis. It was known that epigenetic factors play a role in the pathogenesis of endometriosis, one of which was a change in the level of DNA methylation. This study aims to assess the level of methylation of the PAI-1 gene and its relationship to the development of ovarian and peritoneal endometriosis tissue. This cross-sectional study used a sample of 13 women with ovarian endometriosis, 11 women with peritoneal endometriosis, and 11 women without endometriosis. DNA from the sample was isolated, followed by bisulfite conversion, and finally, observation of DNA methylation was done by using methylation-specific polymerase chain reaction (MSP). The results data obtained previously was analyzed by the Kruskal-Wallis test followed by the Mann-Whitney test. There was a significant difference in the level of PAI-1 gene methylation in ovarian endometriosis, peritoneal endometriosis, and control (p = 0,000). This study also found significant differences between ovarian endometriosis and peritoneal endometriosis respectively compared with controls (p = 0.001 and p = 0.000). However, there was no significant difference in the DNA methylation level of the PAI-1 gene in peritoneal endometriosis compared to ovarian endometriosis (p = 0.111). Our study showed low levels of DNA methylation of the PAI-1 gene that could increase PAI-1 gene expression, and this was suggested to contribute to risk factors for endometriosis in the ovaries and peritoneum.
Keywords: DNA methylation, Ovarian endometriosis, PAI-1 gene, Peritoneal endometriosis