miR-3614-5p在卵巢癌细胞中的表达及对卵巢癌细胞增殖和迁移的影响

Expression of miR-3614-5p in Ovarian Cancer Cells and its Effect on the Proliferation and Migration of Ovarian Cancer Cells

目的分析微小RNA(microRNA)-3614-5p在卵巢癌细胞中的表达,探讨其对卵巢癌细胞增殖和迁移的影响及分子机制。方法OncoLnc在线数据库分析miR-3614-5p表达水平与卵巢癌患者生存期的关系。采用实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-3614-5p在卵巢癌细胞系(SKOV-3、OVCAR-3、A2780、HO-8910、OC3)和正常卵巢上皮细胞系(IOSE80)中的表达。以miR-3614-5p表达最低的细胞系为转染对象,分别转染化学合成的miR-3614-5p模拟物(miR-3614-5p组)和miR-NC(对照组)。分别采用CCK-8法和Transwell迁移实验检测高表达miR-3614-5p对细胞增殖和迁移能力的影响。采用生物信息学和双荧光素酶报告基因法预测并验证miR-3614-5p的靶基因。qRT-PCR和Western blot法检测miR-3614-5p对靶基因表达的影响。结果miR-3074-5p表达水平较高的卵巢癌患者生存期长于miR-3074-5p表达水平较低的患者(P<0.05)。与IOSE80细胞比较,miR-3614-5p在卵巢癌细胞系中的表达显著减少(P<0.01),其在OVCAR-3细胞中表达最少(P<0.01)。与对照组比较,miR-3614-5p组OVCAR-3细胞增殖能力显著降低(P<0.05)、细胞迁移能力显著降低(P<0.01)。miR-3614-5p与G蛋白α抑制剂2(G protein alpha inhibiting activity polypeptide 2,GNAI2)mRNA的3′非翻译区存在结合位点(P<0.01)。高表达miR-3614-5p可显著干扰GNAI2基因的表达(P<0.01)。结论miR-3614-5p在卵巢癌细胞系中的表达降低,其可通过干扰靶基因GNAI2的表达,抑制卵巢癌细胞的增殖和迁移过程,miR-3614-5p可能是治疗卵巢癌的新靶点。

Objective To analyze the expression of microRNA-3614-5p in ovarian cancer cells,and explore its effect on the proliferation and migration of ovarian cancer cells and its molecular mechanism.Methods The OncoLnc online database was used to analyze the relationship between the expression level of miR-3614-5p and the survival time of patients with ovarian cancer.qRT-PCR was used to detect the expression of miR-3614-5p in ovarian cancer cell lines(SKOV-3,OVCAR-3,A2780,HO-8910,OC3)and normal ovarian epithelial cell lines(IOSE80).The cell line with the lowest expression of miR-3614-5p was used as the transfection target,and the chemically synthesized miR-3614-5p mimic(miR-3614-5p group)and miR-NC(control group)were transfected respectively.The CCK-8 method and the Transwell migration test were used to detect the effects of highly expressed miR-3614-5p on cell proliferation and migration.Bioinformatics and dual-luciferase reporter gene methods were used to predict and verify the target gene of miR-3614-5p,respectively.qRT-PCR and Western blot were used to detect the effect of miR-3614-5p on target gene expression.Results The survival time of ovarian cancer patients with higher miR-3074-5p expression level was longer than that of patients with lower miR-3074-5p expression level(P<0.05).Compared with IOSE80 cells,the expression of miR-3614-5p in ovarian cancer cell lines was significantly reduced(P<0.01),and its expression in OVCAR-3 cells was the least(P<0.01).Compared with the control group,the proliferation ability of OVCAR-3 cells in the miR-3614-5p group was significantly reduced(P<0.05),and the cell migration ability was significantly reduced(P<0.01).There are binding sites in the 3′untranslated region of miR-3614-5p and G protein alpha inhibiting activity polypeptide 2(GNAI2)mRNA(P<0.01).High expression of miR-3614-5p can significantly interfere with the expression of GNAI2 gene(P<0.01).Conclusion The expression of miR-3614-5p in ovarian cancer cell lines is reduced.It can inhibit the proliferation and migration of ovarian cancer cells by interfering with the expression of the target gene GNAI2.miR-3614-5p may be a new target for the treatment of ovarian cancer.