敲减ARHGAP30基因对宫颈癌Siha细胞增殖及凋亡的影响  

Effect of knockdown of ARHGAP30 on proliferation and apoptosis of Siha cells

作　　者：彭雅婷 刘端 孟洁 李文超 李慧琦 郭华[1] 牛美兰[2] 秦巧红[1] PENG Ya-ting;LIU Duan;MENG Jie;LI Wen-chao;LI Hui-qi;GUO Hua;NIU Mei-lan;QIN Qiao-hong(Dept of Obstetrics and Gynecology Oncology,the Third Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Medical College of Huanghe Science&Technology University,Zhengzhou 450063,China)

机构地区：[1]郑州大学第三附属医院妇科肿瘤科,河南郑州450052 [2]黄河科技学院医学院,河南郑州450063

出　　处：《中国药理学通报》2024年第5期847-853,共7页Chinese Pharmacological Bulletin

基　　金：河南省医学科技攻关计划项目(No 2022013)。

摘　　要：目的探究敲减Rho鸟苷三磷酸酶激活蛋白30(Rho GTPase-activating protein 30,ARHGAP30)后,宫颈癌Siha细胞增殖及凋亡的变化。方法设计特异性shARHGAP30引物并连接pLKO.1载体,转化到大肠杆菌感受态细胞中,再与慢病毒辅助质粒转入HEK-293T细胞,收集细胞上清获得的病毒过滤后感染Siha细胞,RT-qPCR和Western blot检测敲减效率,以及转染后Bax及Bcl-2的表达变化;CCK-8法检测敲减后细胞的增殖水平。结果成功构建敲减ARHGAP30基因的慢病毒质粒,并建立Siha稳转细胞,ARHGAP30在Siha细胞中的转录和翻译减少(P<0.01),Bax/Bcl-2明显降低(P<0.01),凋亡减少,细胞增殖水平升高(P<0.01)。结论ARHGAP30参与Siha细胞的增殖与凋亡,调控ARHGAP30基因或将干扰宫颈癌的发生和发展。Aim To investigate the changes in the proliferation and apoptosis of Siha cells after knocking down Rho GTPase-activating protein 30(ARHGAP30).Methods After designing specific shARHGAP30 primers and connecting them to the pLKO.1 vector,we transformed them into Escherichia coli competent cells,then co-transfecting them with lentiviral helper plasmids into HEK-293T cells.We collected and filtered cell supernatant to obtain the virus to infect Siha cells.RT-qPCR and Western blot were used to detect knockdown efficiency,as well as changes in the expression of Bax and Bcl-2 after transfection.The CCK-8 method was employed to measure the proliferation level of cells after knockdown.Results After successful construction of a lentiviral plasmid with knockdown of the ARHGAP30 gene and establish-ment of stably transfected Siha cells,ARHGAP30 transcription and translation(P<0.01)in Siha cells decreased,Bax/Bcl-2 significantly decreased(P<0.01),indicating decreased apoptosis and increased cell proliferation(P<0.01).Conclusions This study suggests the involvement of ARHGAP30 in the proliferation and apoptosis of Siha cells,and regulating the ARHGAP30 gene may interfere with the occurrence and development of cervical cancer.

关 键 词：SIHA细胞 ARHGAP30 宫颈癌 基因敲减 细胞增殖 细胞凋亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学] R394.2R737.33