miR-133a-5p调控细胞焦亡抑制宫颈癌细胞增殖、迁移和侵袭

miR-133a-5p inhibits cervical cancer cell proliferation,migration,and invasion by regulating pyroptosis

目的:研究miR-133a-5p靶向半胱天冬酶3(caspase-3)调控细胞焦亡对宫颈癌细胞增殖、迁移和侵袭的作用机制。方法:以Hela细胞为研究对象并分为Control组(正常培养宫颈癌Hela细胞)、mimic-NC组(宫颈癌Hela细胞转染mimic-NC)和miR-133a-5p mimic组(宫颈癌Hela细胞转染miR-133a-5p mimic过表达);利用CCK-8检测细胞增殖,染色法检测细胞迁移和细胞侵袭,ELISA检测活性氧(ROS)水平,流式细胞术检测细胞凋亡水平,蛋白质印迹法检测caspase-3、cleaved caspase-3、GSDME和GSDME-NT蛋白的表达水平,实时荧光定量聚合酶链反应检测caspase-3和miR-133a-5p的mRNA表达水平。结果:对照组和mimic-NC组的细胞增殖、迁移和侵袭结果对比没有明显差异,而miR-133a-5p mimic组细胞增殖、迁移和侵袭结果相较于mimic-NC组明显被抑制(均P<0.05);流式细胞术检测结果发现miR-133a-5p mimic组细胞凋亡水平明显高于mimic-NC组(P<0.05);ELISA检测结果发现miR-133a-5p mimic组ROS水平明显高于mimic-NC组(P<0.05);Western Blot和PCR检测结果发现与mimic-NC组相比,过表达miR-133a-5p能够明显提升cleaved caspase-3和GSDME-NT的表达,降低caspase-3和GSDME的表达(P<0.05)。结论:miR-133a-5p对Hela细胞增殖、迁移和侵袭的抑制作用或与靶向调控caspase-3来调节Hela细胞焦亡相关,对caspase-3/GSDME通路的干预有望成为宫颈癌治疗的新方向。

Objective:To investigate the mechanism of miR-133a-5p targeting caspase-3 to regulate pyroptosis on the proliferation,migration and invasion of cervical cancer cells.Methods:Hela cells were divided into three groups:Control group(normal cultured cervical cancer Hela cells),mimic-NC group(cervical cancer Hela cells transfected with mimic-NC)and miR-133a-5p mimic group(cervical cancer Hela cells transfected with miR-133a-5p mimic overexpression).CCK-8 was used to detect cell proliferation,staining was used to detect cell migration and invasion,ELISA was used to detect the level of reactive oxygen species(ROS),flow cytometry was used to detect the level of apoptosis, Western Blot was used to detect the expression of caspase-3,cleaved caspase-3,GSDME and GSDME-NT,real-time fluorescence quantitative polymerase chain reaction was used to detect the mRNA expression of caspase-3 and miR-133a-5p.Results:There was no significant difference in the results of cell proliferation,migration and invasion between the control group and mimic-NC group,but the results of cell proliferation,migration and invasion in miR-133a-5p mimic group were significantly inhibited compared with those in mimic-NC group(P<0.05).Flow cytometry showed that the level of apoptosis in miR-133a-5p mimic group was significantly higher than that in mimic-NC group(P<0.05).The results of ELISA detection showed that the level of ROS in miR-133a-5p mimic group was significantly higher than that in mimic-NC group(P<0.05),and the results of Western Blot and PCR showed that overexpression of miR-133a-5p could significantly increase the expression of cleaved caspase-3 and GSDME-NT and decrease the expression of caspase-3 and GSDME compared with mimic-NC group(P<0.05).Conclusion:The inhibitory effect of miR-133a-5p on the proliferation,migration and invasion of Hela cells may be related to the targeted regulation of caspase-3 to regulate the pyroptosis of Hela cells.The intervention of caspase-3/GSDME pathway is expected to become a new direction of cervical cancer therapy.