miR-369-3p和COL11A1对卵巢癌细胞增殖 迁移及侵袭的影响  

Effects of miR-369-3p and COL11A1 on proliferation, migration, and invasion of ovarian cancer cells

作　　者：徐玉红 张慧雅[1] 郑伟平[1] 阮雅文[1] 卢琪芸[1] 陈君霞[1] XU Yu-Hong;ZHANG Hui-Ya;ZHENG Wei-Ping(Department of Gynecology,Shaoxing People's Hospital,Shaoxing Hospital of Zhejiang University,Shaoxing,Zhejiang 312000,China)

机构地区：[1]绍兴市人民医院妇科浙江大学绍兴医院妇科,浙江绍兴312000

出　　处：《中国妇幼保健》2021年第18期4327-4331,共5页Maternal and Child Health Care of China

基　　金：浙江省绍兴市公益性应用研究计划项目(2017B7002);绍兴市人民医院青年科研项目(2019YB17)。

摘　　要：目的探讨miR-369-3p和COL11A1对卵巢癌细胞增殖、迁移、侵袭及凋亡的影响,并分析其分子机制。方法实时定量PCR(qRT-PCR)检测正常卵巢上皮细胞IOSE80和卵巢癌细胞SKOV-3中miR-369-3p、COL11A1 mRNA的水平,在卵巢癌SKOV-3细胞中转染miR-369-3p模拟物和pcDNA-COL11A1以过表达miR-369-3p、COL11A1,CCK8法和Transwell实验分别检测卵巢癌SKOV-3细胞增殖、迁移及侵袭能力,蛋白印迹实验(Western blot)检测细胞中COL11A1、Cyclin D1、p21、MMP-2及MMP-9蛋白水平,双荧光素酶报告系统验证miR-369-3p与COL11A1的调控关系。结果与正常卵巢上皮细胞IOSE80(1.00±0.05、1.01±0.08及0.28±0.03)相比,卵巢癌细胞SKOV-3中的miR-369-3p水平(0.41±0.04)显著降低,COL11A1 mRNA(3.26±0.31)和蛋白(0.64±0.06)水平显著升高,差异均有统计学意义(t=21.083、27.643及16.100,P<0.05)。过表达miR-369-3p可降低Cyclin D1、MMP-2及MMP-9蛋白表达量,提高p21表达量,降低细胞OD值、迁移及侵袭细胞个数;miR-369-3p靶向负调控COL11A1的表达;过表达COL11A1可逆转miR-369-3p过表达对SKOV-3细胞增殖、侵袭及迁移的影响。结论 miR-369-3p通过靶向COL11A1调控SKOV-3细胞增殖、迁移及侵袭。miR-369-3p是卵巢癌潜在分子靶点。Objective To explore the effects of miR-369-3 p and COL11 A1 on proliferation, migration, invasion, and apoptosis of ovarian cancer cells, analyze the molecular mechanism. Methods Real-time quantitative PCR was used to detect the levels of miR-369-3 p and COL11 A1 mRNA in normal ovarian cell line IOSE80 and ovarian cancer cell line SKOV-3;miR-369-3 p simulant and pcDNA-COL11 A1 were transfected into ovarian cancer SKOV-3 cells to obtain overexpression of miR-369-3 p and COL11 A1, CCK8 assay and Transwell assay were used to detect proliferation ability, migration ability, and invasion ability of ovarian cancer SKOV-3 cells;Western blotting was used to detect the levels of COL11 A1, Cyclin D1, p21, MMP-2, and MMP-9 proteins;dual-luciferase reporter system was used to verify the regulating relationship between miR-369-3 p and COL11 A1. Results Compared with normal ovarian epithelial IOSE80 cells(1.00±0.05, 1.01±0.08, and 0.28±0.03), the level of miR-369-3 p in into ovarian cancer SKOV-3 cells(0.41±0.04) decreased significantly, the levels of COL11 A1 mRNA(3.26±0.31) and protein(0.64±0.06) increased significantly, there were statistically significant differences(t=21.083, 27.643, 16.100, P<0.05);overexpression of miR-369-3 p reduced the expression levels of Cyclin D1, MMP-2, and MMP-9 proteins, increased the expression level of p21, reduced OD value, the numbers of migration and invasion cells, miR-369-3 p negatively regulated COL11 A1 expression;overexpression of COL11 A1 reversed the effect of miR-369-3 p overexpression on proliferation, migration, and invasion of ovarian cancer cells. Conclusion miR-369-3 p can regulate proliferation, migration, and invasion of ovarian cancer cells by regulating COL11 A1, miR-369-3 p is a potential molecular target of ovarian cancer.

关 键 词：卵巢癌 SKOV-3 miR-369-3p COL11A1 增殖 迁移 侵袭 

分 类 号：R711.75[医药卫生—妇产科学]