调控细胞分裂周期蛋白6对人类乳头瘤病毒16阳性宫颈癌细胞Caski增殖和凋亡的影响  

Effect of regulating cell division cycle 6 on proliferation and apoptosis of human papillomavirus 16-positive cervical carcinoma Caski cells

作　　者：马聪[1] 韩玉斌[1] 陈云卿[1] 郭文涛 MA Cong;HAN Yu-bin;CHEN Yun-qing;GUO Wen-tao(Department of General Gynecology,the First People′s Hospital of Foshan,Foshan 528000,China;Basic Medical College,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]广东省佛山市第一人民医院普通妇科,佛山市528000 [2]广东医科大学基础医学院,东莞市523808

出　　处：《广西医学》2021年第11期1335-1339,共5页Guangxi Medical Journal

基　　金：广东省佛山市科技创新项目(2018AB002971)。

摘　　要：目的探讨调控细胞分裂周期蛋白6(CDC6)对人类乳头瘤病毒(HPV)16阳性宫颈癌细胞Caski增殖和凋亡的影响。方法检测HPV阴性的宫颈癌C33-A细胞株、HPV16阳性的宫颈癌Caski细胞株和SiHa细胞株、HPV18阳性的宫颈癌HeLa细胞株中CDC6蛋白及mRNA的表达水平。将CDC6小干扰RNA(siRNA)-1、siRNA-2、siRNA-NC分别转染入宫颈癌Caski细胞(分别设为CDC6 siRNA-1组、CDC6 siRNA-2组、siRNA-NC组),并设立空白对照组,24 h和48 h后分别检测各组CDC6 mRNA和蛋白的表达水平,筛选出最佳的siRNA序列。转染后,检测最佳siRNA序列的CDC6 siRNA组、siRNA-NC组、空白对照组的细胞增殖能力、细胞周期及细胞凋亡率。结果(1)Caski细胞、HeLa细胞、SiHa细胞、C33-A细胞的CDC6 mRNA的相对表达量依次降低;C33-A细胞CDC6蛋白的相对表达量低于其他3种细胞,Caski细胞和HeLa细胞CDC6蛋白的相对表达量均高于SiHa细胞(均P<0.05)。(2)CDC6 siRNA-1组CDC6 mRNA及蛋白相对表达量低于CDC6 siRNA-2、siRNA-NC组与空白对照组(均P<0.05),故选用siRNA-1序列进行后续研究。(3)转染48 h后,与siRNA-NC组与空白对照组比较,CDC6 siRNA-1组的细胞增殖能力降低,细胞凋亡率增加,细胞G 1期变长、G 2期缩短(均P<0.05)。结论沉默CDC6能够有效干扰HPV16阳性宫颈癌Caski细胞的增殖,靶向抑制CDC6有望成为早期治疗宫颈癌的新途径。Objective To investigate the effect of regulating cell division cycle 6(CDC6)on the proliferation and apoptosis of human papillomavirus(HPV)16-positive cervical carcinoma Caski cells.Methods Detection of CDC6 mRNA and protein expression levels was conducted in HPV-negative cervical carcinoma C33-A cell line,HPV16-positive cervical carcinoma Caski cell line and SiHa cell line,and HPV18-positive cervical carcinoma HeLa cell line.CDC6 small interfering RNA(siRNA)-1,siRNA-2 and siRNA-NC were transfected into cervical carcinoma Caski cells(defined as CDC6 siRNA-1 group,CDC6 siRNA-2 group and siRNA-NC group,respectively),and the blank control group was established,then CDC6 mRNA and protein expression levels were determined in each group 24 and 48 hours later to screen the optimal siRNA sequence.After transfection,the cell proliferation capacity,cell cycle and cell apoptosis rate were detected in the optimal CDC6 siRNA groups,siRNA-NC group and blank control group.Results(1)The relative expression of CDC6 mRNA decreased in the order of Caski cells,HeLa cells,SiHa cells and C33-A cells;the relative expression of CDC6 protein in C33-A cells was lower than that in the other three cells,and Caski cells and HeLa cells yielded a higher relative expression of CDC6 protein as compared with SiHa cells(all P<0.05).(2)The relative expression of CDC6 mRNA and protein in the CDC6 siRNA-1 group was lower than that in the CDC6 siRNA-2,siRNA-NC and blank control groups(all P<0.05),thus siRNA-1 sequence was involved in follow-up experiments.(3)After 48 hours of transfection,by comparison with the siRNA-NC group and the blank control group,the CDC6 siRNA-1 group possessed attenuated cell proliferation capacity and increased cell apoptosis rate,and exhibited longer G 1 stage and shorter G 2 stage(all P<0.05).Conclusion Silencing CDC6 can effectively suppress the proliferation of HPV16-positive cervical cancer Caski cells,targeted inhibition of CDC6 is a new promising approach to early treatment of cervical cancer.

关 键 词：宫颈癌 细胞分裂周期蛋白6 人类乳头瘤病毒16 CASKI细胞 细胞增殖 细胞周期 细胞凋亡 RNA干扰技术 

分 类 号：R737.33[医药卫生—肿瘤]