中文摘要：

目的检测microRNA-107（miR-107）在卵巢癌中的表达水平,探讨其对卵巢癌细胞系侵袭转移的调控作用及分子机制。方法 qRT-PCR法比较卵巢癌组织、正常卵巢组织、卵巢癌细胞系和正常卵巢上皮细胞中miR-107表达差异,用生物信息学的方法特异预测出miR-107靶基因Dicer1后,通过双荧光素酶报告体系进行验证。上调miR-107表达后,Western blot检测Dicer1的表达变化;划痕实验、Tr-answell实验观察卵巢癌细胞运动和侵袭能力的改变;qRT-PCR及Confocal观察EMT相关分子E-cad-herin、β-Catenin、N-cadherin、Vimentin、Fibronectin、ICAM-1、MMP-9的表达。结果 miR-107在上皮性卵巢癌组织的表达较正常卵巢组织显著升高（P〈0.05）;双荧光素酶报告基因系统验证预测结果正确。上调miR-107后,Western blot结果显示Dicer1蛋白质水平被显著抑制,卵巢癌细胞系SKOV3细胞形态发生间质细胞样转变,间质标志分子β-Catenin、N-cadherin及MMP-9表达上升;体外实验表明卵巢癌细胞的运动和侵袭能力得到明显促进。结论本研究表明miR-107促进卵巢癌侵袭转移的分子机制是通过下调miRNA加工机器Dicer1,从而介导卵巢癌细胞EMT的发生;抑制miR-107或恢复Dicer1水平可能成为上皮性卵巢癌治疗的有效手段。

英文摘要：

Objective To explore the expression level of MicroRNA-107 （miR-107） in ovarian cancer, so as to figure out the mechanism involved in invasion and metastasis control of ovarian cancer cell lines for miR-107. Methods By comparison miR-107 expression levels in ovarian cancer and normal ovarian tis- sues, or between normal ovarian surface epithelium and ovarian cancer cell lines by qRT-PCR, we con firmed the differential expression and then predicted Dicerl as its specific validated target gene with the approach of bioinformatics. Furthermore, the dual luciferase reporter system was adopted to verify the prediction. After elevating miR-107 level, Western blot was performed to detect the protein level of Dic erl ; scratch test and transwell assay were employed to check the alteration of movements, invasion and metastasis in ovarian cancer cell line SKOV3; furthermore, qRT-PCR and Confocal were applied to study the level of EMT associated molecular, such as E-cadherin, -Catenin, N-cadherin, Virnentin, Fibronec- tin, ICAM-1, and MMP-9 respectively. Results The expression level of miR-107 in epithelial ovarian carcinoma was increased when compared with that of normal tissues （P〈0. 05）. The dual luciferase re- porter system validated Dicerl as a specific target gene of miR-107. Dramatically reduction of Dicer1 was observed with miR 107 overexpression, at the same time, morphological change occured and cell invasion and metastasis ability increased a lot due to the upregulation of mesenchymal molecules, including -Cate- nin, N-eadherin, MMP-9 following miR-107 upregulated. Conclusion miR-107, acting as an oncogene miRNA, promoted invasion and metastasis via downregulation of miRNA regulatory machinery Dicer1 and mediated EMT process in ovarian cancer. Inhibition of miR-107 or restoration of Dieerl may repre- sent a new potential therapeutic target for ovarian cancer treatment.