KCTD11对宫颈癌HeLa细胞增殖的影响及分子机制  

Influence and molecular mechanism of KCTD11 on the proliferation of cervical cancer cell HeLa

作　　者：张激 谭兴民[2] ZHANG Ji;TAN Xingmin(Department of Oncology,Sichuan Provincial Hospital of Integrated Traditional Chinese and Western Medicine,Chengdu,Sichuan 610041,China;Department of Gynaecology,Sichuan Provincial Hospital of Integrated Traditional Chinese and Western Medicine,Chengdu,Sichuan 610041,China)

机构地区：[1]四川省中西医结合医院肿瘤科,四川成都610041 [2]四川省中西医结合医院妇科,四川成都610041

出　　处：《国际检验医学杂志》2022年第6期739-744,共6页International Journal of Laboratory Medicine

摘　　要：目的探讨含钾通道四聚结构域蛋白11(KCTD11)对宫颈癌HeLa细胞增殖的影响及分子机制。方法利用数据库比较KCTD11在宫颈癌组织及正常组织中的表达,并分析KCTD11表达与宫颈癌级别的关系。利用短发夹RNA载体构建技术,敲低KCTD11的表达,并利用MTT试验、流式细胞术检测KCTD11敲低对HeLa细胞增殖的影响。采用实时荧光定量PCR(RT-qPCR)和Western blot检测下调KCTD11对细胞周期蛋白mRNA及蛋白水平的影响。同时,利用染色质免疫沉淀技术(ChIP)对其分子机制进行分析。结果KCTD11在宫颈癌组织中的表达水平显著高于其在正常组织中的表达水平(P<0.05)。生长曲线及细胞周期分析结果表明,敲低KCTD11后,细胞周期阻滞于G0/G1期(P<0.05),细胞增殖速率降低(P<0.001)。并导致细胞周期相关蛋白(Myc、CDK2、CDK4、CDK6、CCND3及CCNE2)的mRNA及蛋白水平均下调(P<0.001)。数据库分析结果表明,多个细胞周期蛋白的表达与KCTD11的表达呈正相关。而ChIP分析结果表明,KCTD11能够结合到原癌基因Myc启动子上。结论KCTD11能够通过转录激活原癌基因Myc的表达,上调细胞周期相关蛋白的表达,从而促进宫颈癌细胞的增殖。Objective To investigate the effect and mechanisms of potassium channel tetramerization domain containing 11(KCTD11)on the proliferation of cervical cancer cell HeLa.Methods The expression of KCTD11 in cervical cancer and normal tissue was compared using online database.The relationship between the expression of KCTD11 and the grade of cervical cancer was analyzed using online database.The expression of KCTD11 was knocked down by short-hairpin RNA(shRNA),the effect of KCTD11 knockdown on the proliferation of HeLa cells was detected by flow cytometry and MTT assay.RT-qPCR and Western blot were used to detect the effect of down-regulation of KCTD11 on mRNA and protein expression of cell cycle-related proteins.In addition,chromatin immunoprecipitation assay(ChIP assay)was used to explore the molecular mechanism of KCTD11 effect on cervical cancer cell proliferation.Results The expression of KCTD11 in cervical cancer was significantly higher than that in normal tissues,and the expression level increased significantly with the increase of cervical cancer grade.The results of MTT assay and cell cycle analysis showed that the cell cycle was arrested in G0/G1 phase after KCTD11 knocked down(P<0.05),and cell proliferation rate was significantly reduced(P<0.001).The results of database analysis showed that the expressions of cell cycle-related proteins(Myc,CDK2,CDK4,CDK6,CCND3 and CCNE2)were positively correlated with the expression of KCTD11.In addition,the mRNA and protein levels of cell cycle-related proteins(Myc,CDK2,CDK4,CDK6,CCND3 and CCNE2)were significantly down-regulated with KCTD11 down-regulation(P<0.001).The results of ChIP assay showed that KCTD11 could bind on Myc promoter.Conclusion KCTD11 upregulates the expression of cell cycle-related proteins and promotes cell proliferation of cervical cancer cells by binding on Myc promoter and activating Myc expression.

关 键 词：含钾通道四聚结构域蛋白11 宫颈癌 细胞增殖 MYC 

分 类 号：R737.34[医药卫生—肿瘤]