| 摘要: | 子宮內膜異位是一種婦產科常見的疾病，而造成此疾病的發病過程及機轉並不清楚。近年來有研究指出在大鼠的子宮內膜異位的模式中，發現骨橋蛋白(osteopontin, OPN)在子宮組織內會大量表現。本論文主旨在探討OPN在子宮內膜異位的形成中所扮演的角色。首先，我們利用西方墨點法發現子宮內膜異位病患的子宮內膜組織、腹膜液、濾泡液和巧克力囊腫體液中OPN會大量表現。此外，我們並以子宮內膜HEC-1A 細胞為細胞模式，觀察OPN對子宮內膜細胞的移行、入侵和增生能力的影響。接著我們以不同劑量的OPN去處理細胞，細胞產生移行及入侵，但不具有增生能力。由OPN引發的細胞移行及入侵可被αvβ3 integrin inhibitor所減緩。另一方面，我們發現β-estradiol和 progesterone會增加細胞內OPN表現及分泌型的OPN釋出，也會誘導細胞產生移行，但當β-estradiol和 progesterone共同作用時，OPN表現及細胞移行現象則降低。接著，我們將細胞中內生性的OPN knockdown後，可減緩β-estradiol所誘導細胞移行的現象及OPN的分泌。接下來我們更進一步探討OPN於細胞內的訊息傳遞路徑，當細胞經由1μM Wortmanin (PI-3k inhibitor)前處理後，OPN仍可引發細胞移行且NF-κB蛋白質表現增加，但是在10μM Wortmanin (PI-3k inhibitor)前處理下，移行現象則被抑制。最後我們從zymography assay中發現OPN會促進細胞表現MMP-2、MMP-9。由上述的結果，我們推測OPN可被β-estradiol刺激而表現及分泌釋出，可再經由 autocrine或paracrine的機制，與細胞上αvβ3 integrin結合而促使細胞移行及侵入，其中訊息傳遞路徑可能是經由 PI-3K路徑活化MMP2/9及產生actin重組（remodeling）。
Endometriosis is a common gynecological disease. The pathology and etiology of endometriosis is still unclear. Recently, enhanced osteopontin（OPN） expression was found in the endometriotic tissue of rat model. We propose that OPN could promote the endometriotic cell migration and invasion. The differential proportion of OPN was detected in the endometrium from the patients with endometrial hyperplasia, myoma, endometriosis or adenomyosis. A significant increase of OPN was found in the eutopic endometrium in the women with adenomyosis. In this study, HEC-1A cell was used as the in vitro enodometriotic cell model to illustrate the effects of OPN on cell migration, invasion and proliferation. A dose-response and time-dependent increase of cell migration and cell invasion was demonstrated. But there was no difference in cell proliferation followed the OPN treatment. Nevertheless, the OPN induced cell migration and invasion was attenuated or inhibited by αvβ3 integrin inhibitor (anti-αvβ3 antibody). On the other hand, we also found β-estradiol or progesterone was found to augment OPN expression and secretion, and induced cell migration. β-estradiol-induced cell migration was attenuat followed the silence the OPN expression by OPN siRNA. In immunoflurescent staining, we observed actin remodeling in the-OPN treated cells. We also found a dose response activation of MMP-2 and MMP-9 followed OPN treatment. Finally, we found cell migration was attenuated by Wortmannin (PI-3k inhibitor). We suggested OPN-induced cell migration and invasion was mediated by the OPN-αvβ3 interaction. The molecular signal pathway of OPN/αvβ3-mediated MMP activation and actin remodeling through PI-3K. |
