长链非编码RNA NR2F1-AS1靶向微小RNA-493-5p表达调控卵巢癌细胞增殖及凋亡的机制研究  

Mechanism of long non-coding RNA NR2F1-AS1 regulates proliferation and apoptosis of ovarian cancer cells by targeting microRNA-493-5p expression

作　　者：张羽 张璨 章伟玲[1] ZHANG Yu;ZHANG Can;ZHANG Weiling(Department of Gynecology,Nantong City Cancer Hospital in Jiangsu Province,Nantong,Jiangsu,226361)

机构地区：[1]江苏省南通市肿瘤医院妇科,江苏南通226361

出　　处：《实用临床医药杂志》2023年第4期30-34,共5页Journal of Clinical Medicine in Practice

基　　金：江苏省南通市科技计划项目(JC2018045)。

摘　　要：目的探讨长链非编码RNA NR2F1-AS1(LncRNA NR2F1-AS1)靶向微小RNA-493-5p(miR-493-5p)的表达调控卵巢癌细胞增殖及凋亡的机制。方法采用实时荧光定量聚合酶链反应(qRT-PCR)检测NR2F1-AS1与miR-493-5p在不同卵巢癌细胞株中的表达水平;采用双荧光素酶报告基因检测NR2F1-AS1与miR-493-5p的相互作用;采用MTT增殖实验检测干扰NR2F1-AS1表达或miR-493-5p过表达后卵巢癌细胞增殖能力的变化;采用流式细胞术检测细胞凋亡率;采用Western blot检测CyclinD1、p21、p27、Bcl-2、Bax、cleaved-caspase 3蛋白表达。结果与正常卵巢上皮细胞比较,NR2F1-AS1在卵巢癌细胞中的表达水平升高,而miR-493-5p的表达水平则降低,差异有统计学意义(P<0.05)。双荧光素酶实验证实NR2F1-AS1能与miR-493-5p特异性结合,并可负性调控miR-493-5p的表达及活性。干扰NR2F1-AS1表达或miR-493-5p过表达后可抑制卵巢癌细胞增殖能力,促进细胞凋亡,并可显著上调p21、Bax、p27、cleaved-caspase 3蛋白的表达,下调CyclinD1、Bcl-2蛋白的表达。抑制miR-493-5p表达可逆转干扰NR2F1-AS1表达对卵巢癌细胞增殖及凋亡的作用。结论干扰NR2F1-AS1表达可通过上调miR-493-5p的表达抑制卵巢癌细胞增殖及诱导细胞凋亡。Objective To explore the mechanism of long noncoding RNA NR2F1-AS1(LncRNA NR2F1-AS1)in regulating proliferation and apoptosis of ovarian cancer cells by targeting microRNA-493-5p(miR-493-5p)expression.Methods The quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of NR2F1-AS1 and miR-493-5p in different lines of ovarian cancer cells;the dual luciferase reporter gene was used to detect the interaction between NR2F1-AS1 and miR-493-5p;the MTT proliferation assay was used to determine the change in the proliferation ability of ovarian cancer cells after interference with NR2F1-AS1 expression or miR-493-5p overexpression;the apoptosis rate of ovarian cancer cells was detected by flow cytometry;the Western blot was used to detect the protein expression levels of CyclinD1,p21,p27,Bcl-2,Bax and cleaved-caspase 3.Results Compared with normal ovarian epithelial cells,the expression level of NR2F1-AS1 in ovarian cancer cells increased significantly,while the expression level of miR-493-5p decreased significantly(P<0.05).The dual luciferase assay confirmed that NR2F1-AS1 could specifically bind to miR-493-5p,and the expression and activity of miR-493-5p were negatively regulated by NR2F1-AS1.Interference with NR2F1-AS1 expression or overexpression of miR-493-5p could inhibit the proliferation of ovarian cancer cells,promote apoptosis,significantly up-regulate the expression of p21,Bax,p27 and cleaved-caspase 3,and down-regulate protein expression levels of CyclinD1 and Bcl-2.Inhibiting the expression of miR-493-5p was able to reverse the effect of interference with[JP3]the expression of NR2F1-AS1 on the proliferation and apoptosis of ovarian cancer cells.Conclusion Interference with the expression of NR2F1-AS1 can inhibit the proliferation of ovarian cancer cells and induce apoptosis by up-regulating the expression of miR-493-5p.

关 键 词：卵巢癌 长链非编码RNA NR2F1-AS1 微小RNA493-5p 增殖 凋亡 

分 类 号：R737.31[医药卫生—肿瘤] Q344[医药卫生—临床医学]