FOXF1-AS1负调控miR-146b-3p对宫颈癌细胞增殖凋亡的影响

Effects of FOXF1 AS1 on proliferation and apoptosis of cervical cancer cells by negatively regulating miR-146b-3p

目的探讨FOXF1-AS1对宫颈癌细胞增殖凋亡的影响及分子机制。方法选取2017年5月至2019年12月确诊的35例宫颈癌患者的癌组织和癌旁组织;宫颈癌细胞SiHa分为pcDNA组、pcDNA-FOXF1-AS1组、pcDNA-FOXF1-AS1+miR-NC组、pcDNA-FOXF1-AS1+miR-146b-3p组。实时荧光定量PCR(RT-qPCR)检测FOXF1-AS1和miR-146b-3p表达水平;四甲基偶氮唑盐比色法(MTT)检测细胞增殖;克隆形成实验检测细胞克隆形成数;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测蛋白表达;双荧光素酶报告实验检测FOXF1-AS1和miR-146b-3p的靶向关系。结果与癌旁组织组相比,宫颈癌组织中FOXF1-AS1表达水平显著降低(P<0.05)。过表达FOXF1-AS1后,SiHa细胞中miR-146b-3p表达水平降低,细胞OD值降低,细胞克隆形成数减少,细胞凋亡率升高,Cleaved-caspase3表达水平升高,pro-caspase3表达水平降低(P<0.05)。FOXF1-AS1靶向调控miR-146b-3p,miR-146b-3p可逆转过表达FOXF1-AS1对SiHa增殖凋亡的影响。结论过表达FOXF1-AS1可能通过下调miR-146b-3p抑制宫颈癌SiHa细胞增殖,促进细胞凋亡。

Objective To investigate the effects of FOXF1-AS1 on the proliferation and apoptosis of cervical cancer cells and its molecular mechanism.Methods The cancer tissues and para-carcinoma tissues of 35 patients with cervical cancer who were diagnosed in our hospital from May 2017 to December 2019 were selected.The cervical cancer cells SiHa were divided into pcDNA group,pcDNA-FOXF1-AS1 group,pcDNA-FOXF1-AS1+miR-NC group,pcDNA-FOXF1-AS1+miR-146b-3p group.Real time fluorescent quantitative PCR(RT qPCR)was used to detect the expression levels of FOXF1-AS1 and miR-146b-3p.The tetramethylazolium salt colorimetric method(MTT)was used to detect cell proliferation.The clone formation experiment was used to detect the number of cell clone formation.The cell apoptosis was detected by flow cytometry,and the protein expression was detected by Western Blot.Moreover the dual luciferase reporter experiment was used to detect the targeting correlation between FOXF1-AS1 and miR-146b-3p.Results As compared with those in adjacent tissues,the expression levels of FOXF1-AS1 in cervical cancer tissue were significantly decreased(P<0.05).After overexpression of FOXF1-AS1,the expression levels of miR-146b-3p in SiHa cells were decreased,cell OD value was decreased,cell clone formation number was decreased,cell apoptosis rate was increased,and the expression levels of Cleaved Caspase 3 were increased,however,those of pro-caspase3 expression were significantly decreased(P<0.05).FOXF1-AS1 targetedly regulating miR-146b-3p,miR-146b-3p could reverse the effects of overexpression of FOXF1-AS1 on the proliferation and apoptosis of SiHa cells.Conclusion The overexpression of FOXF1-AS1 may inhibit the proliferation of cervical cancer SiHa cells and promote cell apoptosis by down-regulating the expression of miR-146b-3p.