长链非编码RNA ARAP1-AS1在卵巢癌中的表达及作用机制研究

Expression and mechanism of long noncoding RNA ARAP1-AS1 in ovarian cancer

目的从分子生物学角度阐述长链非编码RNA(lncRNA)ARAP1反义AS1(ARAP1-AS1)的表达水平在卵巢癌细胞生长分化中的作用及机制,从而揭示lncRNA ARAP1-AS1对卵巢癌恶性程度、生长速度及侵袭能力的影响。方法分别购买人正常卵巢表面上皮细胞系和卵巢癌细胞系进行培养和传代。采用实时定量聚合酶链反应(RT-PCR)对卵巢癌细胞及正常卵巢细胞中lncRNA ARAP1-AS1表达水平进行检测和评价;采用细胞计数试剂盒-8(CCK-8)、溴脱氧核苷尿嘧啶和流式细胞术检测细胞增殖和凋亡;通过Western blot检测miR-4735-3p/多态腺样瘤蛋白2(PLAGL2)信号通路相关蛋白表达变化;使用Transwell分析验证细胞迁移和侵袭。结果RT-PCR结果显示,ARAP1-AS1在卵巢癌细胞系中呈现高表达(P<0.05)。ARAP1-AS1的欠表达可以抑制卵巢癌细胞增殖、迁移和侵袭(P<0.01)。同时沉默ARAP1-AS1可以抑制miR-4735-3p/PLAGL2信号通路相关蛋白的表达水平。结论ARAP1-AS1通过miR-4735-3p基因增强了PLAGL2的表达,即ARAP1-AS1通过miR-4735-3p/PLAGL2信号通路在卵巢癌中发挥致癌作用。

Objective To elucidate the role and mechanism of the expression level of lncRNA ARAP1-AS1 in the growth and differentiation of ovarian cancer cells from the perspective of molecular biology,in order to reveal the impact of lncRNA ARAP1-AS1 on the malignancy,growth rate and invasion ability of ovarian cancer.Methods Human normal ovarian surface epithelial cell lines and ovarian cancer cell lines were purchased for culture and passage.Real-time quantitative polymerase chain reaction(RT-PCR)was used to detect and evaluate the expression level of lncRNA ARAP1-AS1 in ovarian cancer cells and human normal ovarian cells.Cell proliferation and apoptosis were detected by cell counting kit-8(CCK-8),bromodeoxynucleoside uracil and flow cytometry.The expression of miR-4735-3p/PLAGL2 signaling pathway related proteins was detected by Western blot.Cell migration and invasion were verifted by Transwell analysis.Results RT-PCR results showed that ARAP1-AS1 was high expressed in ovarian cancer cell lines(P<0.05).The underexpression of ARAP1-AS1 could inhibit ovarian cancer cells proliferation,migration and invasion(P<0.01).At the same time,silencing ARAP1-AS1 could inhibit the expression level of miR-4735-3p/PLAGL2 signaling pathway related proteins.Conclusion ARAP1-AS1 enhances the expression of PLAGL2 through the miR-4735-3p gene.ARAP1-AS1 plays a carcinogenic role in ovarian cancer through the miR-4735-3p/PLAGL2 signaling pathway.