目的：探讨miR-139-5p靶向Notch1抑制上皮性卵巢癌（epithelial ovarian cancer，EOC）细胞增殖和侵袭的作用机制。方法：选取2018年1月至2018年12月在河南省南阳市中心医院妇科手术切除的24例EOC患者的癌和相应的癌旁组织标本，以及人卵巢癌细胞系SKOV3、ES2、HEY-T30和人卵巢上皮细胞株IOSE80，用qPCR检测EOC组织和细胞系中miR-139-5p和Notch1 mRNA的表达。将过表达miR-139-5p载体、重组质粒pLV-Notch1转染至SKOV3细胞，并设置空白对照组（Ctrl组）和阴性对照组（NC组），用双荧光素酶报告基因实验验证miR-139-5p与Notch1 3’-UTR靶向关系，用CCK-8、Transwell、划痕愈合实验分别检测细胞的增殖、侵袭和迁移能力，用Western blotting检测细胞中增殖和迁移相关蛋白的表达。结果：与癌旁组织和IOSE80细胞比较，EOC组织和细胞系中miR-139-5p表达显著降低、Notch1 mRNA表达显著升高（均P<0.01）。双荧光素酶报告基因实验结果证实，Notch1是miR-139-5p的靶基因。与NC组比较，miR-139-5p mimic组3 d时SKOV3细胞的增殖、侵袭、迁移能力和Notch1、NICD、Cyclin D1、Cyclin A1、Snail1、β-catenin及N-cadherin表达水平均明显降低（均P<0.01），E-cadherin表达水平明显升高（P<0.01）；同时过表达Notch1可逆转miR-139-5p抑制SKOV3细胞增殖、侵袭与迁移的作用。结论：miR-139-5p可靶向Notch1抑制EOC细胞的增殖、侵袭和迁移能力，可能与其下调NICD、Cyclin D1、Cyclin A1、Snail1、β-catenin、N-cadherin而上调E-cadherin的表达水平有关。

Objective: To explore the action mechanism of miR-139-5p inhibiting proliferation and invasion of epithelial ovarian cancer (EOC) cells by targetedly regulating Notch1. Methods: A total of 24 pairs of EOC tissues and its corresponding para-cancerous tissues from patients, who underwent surgical resection in the Department of Gynecology, Nanyang Central Hospital of Henan Province,were collected for this study; in addition, human ovarian cancer cell lines (SKOV3, ES2, HEY-T30) and human ovarian epithelial cell line IOSE80 were also collected. Real-time quantitative PCR (qPCR) was applied to detect mRNAexpression of miR-139-5p and Notch1 in EOC tissues and cell lines. The miR-139-5p over-expression vector and recombinant plasmid pLV-Notch1 were transfected into SKOV3 cells. Blank control group (Ctrl group) and negative control group (NC group) were set up. Dual luciferase reporter gene assay was applied to verify the targeting relationship between miR-139-5p and Notch1 3'-UTR. CCK-8, Transwell and Scratch healing experi‐ments were applied to detect cell proliferation invasion and migration, respectively. Western blotting was applied to detect expressions of proliferation and migration related proteins in cells. Results: Compared with para-cancerous tissues and IOSE80 cells, the expression of miR-139-5p was significantly decreased in EOC tissues and cell lines, while the expression of Notch1 mRNA was significantly in‐creased (all P<0.01). The results of Dual luciferase reporter showed that Notch1 was the downstream target gene of miR-139-5p. Com‐pared with NC group, cell proliferation, invasion and migration ability, expression levels of Notch1, NICD, Cyclin D1, Cyclin A1,Snail1, β-catenin and N-cadherin were all significantly decreased on 3 d in miR-139-5p mimic group (all P<0.01), while expression of E-cadherin was significantly increased (P<0.01); meanwhile, over-expression of Notch1 could reverse the inhibitory effect of miR-139- 5p on proliferation, invasion and migration of SKOV3 cells. Conclusion: miR-139-5p can targetedly regulate Notch1 to inhibit prolifer‐ation, invasion and migration of EOC cells, which may be related to its down-regulation of NICD, Cyclin D1, Cyclin A1, Snail1, β-catenin and N-cadherin, and up-regulation of E-cadherin.

河南省医学科技攻关计划资助项目（No.201503118）