摘要
目的miR-425-5p对宫颈癌HeLa细胞功能的调控作用及其机制。方法将体外培养的HeLa细胞分为miR-NC组(转染miR-425-5p模拟物对照)、miR-425-5p组(转染miR-425-5p模拟物)、anti-miR-NC组(转染miR-425-5p抑制剂对照)和anti-miR-425-5p组(转染miR-425-5p抑制剂)共四组。采用RT-PCR检测四组细胞中miR-425-5p的表达水平,CCK-8法、克隆形成试验、Transwell试验、划痕试验和流式细胞仪分别检测细胞增殖、克隆形成、侵袭、迁移和凋亡能力。生物信息学软件预测、双荧光素酶报告基因试验检测miR-425-5p和磷酸酯酶与张力蛋白同源物(PTEN)的靶向关系,Western blot检测miR-425-5p对PTEN蛋白表达的影响。结果转染miR-425-5p模拟物后HeLa细胞中miR-425-5p的表达水平较miR-NC组明显升高,而miR-425-5p抑制剂后miR-425-5p的表达水平较anti-miR-NC组明显降低,差异均具有统计学意义(均P<0.05)。与miR-NC组相比,miR-425-5p组中细胞OD值、克隆形成率、迁移率和侵袭细胞数均明显升高,而细胞凋亡率明显降低,差异均具有统计学意义(均P<0.05);与anti-miR-NC组相比,anti-miR-425-5p组中细胞OD值、克隆形成率、迁移率和侵袭细胞数均明显降低,而细胞凋亡率明显升高,差异均具有统计学意义(均P<0.05)。双荧光素酶报告基因试验证实miR-425-5p能够与PTEN 3′UTR靶向结合,Western blot试验结果证实miR-425-5p可负向调控PTEN蛋白的表达。结论miR-425-5p可促进HeLa细胞增殖、侵袭、迁移并抑制细胞凋亡,其作用机制可能与靶向调控PTEN表达有关。
Objective To investigate the effect of microRNA-425-5 p(miR-425-5 p)on the function of HeLa cells in cervical cancer and its mechanism.Methods HeLa cells cultured in vitro were divided into miR-NC group(transfected miR-425-5 p mimic control),miR-425-5 p group(transfected miR-425-5 p mimic),anti-miR-NC group(transfected with miR-425-5 p inhibitor control)and anti-miR-425-5 p group(transfected with miR-425-5 p inhibitor).The expression level of miR-425-5 p in each group of cells was detected by RT-PCR,and cell proliferation,colony formation,invasion,migration and apoptosis ability were detected by CCK-8 method,colony formation assay,Transwell assay,scratch assay and flow cytometry separately.Bioinformatics software predicted and dual luciferase reporter gene assay detected the targeting relationship of miR-425-5 p and phosphatase and tensin homolog(PTEN),and the effect of miR-425-5 p on PTEN protein expression was measured by Western blot.Results The expression level of miR-425-5 p in HeLa cells transfected with miR-425-5 p mimic was significantly higher than that in the miR-NC group,while the expression level of miR-425-5 p was significantly lower in the miR-425-5 p inhibitor group than that in the anti-miR-NC group,with statistically significant differences(all P<0.05).Compared with the miR-NC group,the OD value,clone formation rate,migration rate and invasive cell number in the miR-425-5 p group increased significantly,while the apoptotic rate decreased significantly,with statistically significant differences(all P<0.05).Compared with the anti-miR-NC group,the OD value,clone formation rate,migration rate and invasive cell number in the anti-miR-425-5 p group decreased significantly,while cell apoptosis rate increased significantly,with statistically significant differences(all P<0.05).Dual luciferase reporter gene experiment confirmed that miR-425-5 p could target PTEN 3′UTR.Western blot results confirmed that miR-425-5 p could negatively regulate PTEN protein expression.Conclusions MiR-425-5 p can promote the proliferation,invasion,migration and inhibit apoptosis of HeLa cells,and its mechanism may be related to the targeting regulation of PTEN expression.
作者
姚伟林
岳红萍
YAO Weilin;YUE Hongping(Department of Gynecology,Yunnan Third People's Hospital,Kunming 650011,Yunnan,China)
出处
《中国性科学》
2020年第8期10-14,共5页
Chinese Journal of Human Sexuality
