| Summary: | 碩士 === 臺北醫學大學 === 醫學科學研究所 === 98 === Endometriosis is the growth of endometrial tissues eutopically and/or ectopically. This disease affects nearly 10% of reproductive aged women with symptoms of persistent pelvic pain, abnormal uterine bleeding, and infertility. Stem cells in endometrial basalis layer have been hypothesized to mediate endometrium regeneration as well as the initiation of endometriosis. However, mechanism involved in endometriosis transformation processes still remains largely unknown. In clinic observation, chronic inflammation is tightly associated with endometriosis. Several cytokines including interleukin (IL)-1, 6, 8, 10, tumor necrosis factor alpha (TNF-α), and vascular endothelial growth factor (VEGF) have been reported to be increased in the peritoneal fluid (PF) of women with endometriosis. In this aspect, we hypothesized that inflammation may result in transformation of endometrium stem cells and initiate the processes of endometriosis. To test this hypothesis, tissues with hyperplasia (n = 37), endometriosis (n = 43), and adenomyosis (n = 8) were collected and the clinical CA125 score was examined. Tissues were then analyzed to correlate with the pluripotent gene expression (Oct-4 and Nanog) with endometriosis progression in human. In our results, while comparison with the hyperplasia tissues, tissues with endometriosis/adenomyosis showed a higher expression of Oct-4 and Nanog as well as correlation with CA125. Furthermore, we established a serum-free culture system to generate mouse epithelial-like endometrium stem cells (mEESCs). These mEESCs showed a strong expression of stem cell markers (such as Oct-4 and SSEA-1) as well as IGF-1/IGF-1R (stemness regulatory signaling); and a weak expression of CD49f, CD34, CD133, α-SMA, and p63. Interestingly, by real-time PCR analysis, the mEESCs showed an increasing of stemness (such as Oct-4, Sox2, c-Myc, Klf-4, and IGF-1) under the treatment of inflammation condition medium (CM). Under the inflammation treatment, a down-regulation of E-cadherin transcription along with the up-regulation of N-cadherin transcription was observed, suggesting EMT processes of mEESCs. Furthermore, to investigate inflammation effect on the RL95-2 cell line (low Oct-4 expression endometrium carcinoma cell line), we transfected a Promotor-Oct-4-EGFP plasmid and a HA-Oct-4 plasmid individually into RL95-2 cell line. A higher GFP expression was observed under the inflammation treatment, indicating a stronger Oct-4 expression. Also, a higher Oct-4 and HA protein expression were detected by western blot in the HA-Oct-4 RL95-2 cell line under the inflammation condition. An increasing expression of Oct-4 and N-cadherin mRNA were showed in the HA-Oct-4 RL95-2 cell line. These results suggested increasing of stemness characteristic and EMT processes under treatment of the inflammation condition medium. In conclusion, we demonstrated that chronic inflammation processes might play an important role in initiation and development of endometriosis by transforming of endometrium epithelial stem cells. This observation may provide the possible therapeutic target of infertility and tumorigenesis.
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