GTF2H2的核苷酸切除修复功能对肝癌细胞增殖和凋亡的影响  

Effects of NER function of GTF2H2 on proliferation and apoptosis of liver cancer cells

作　　者：周冬虎 李艳萌[1] 贾思雨 王何静 齐赛平 黄坚[1] ZHOU Dong-hu;LI Yan-meng;JIA Si-yu(Experimental Center,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院科研实验中心北京市临床医学研究所

出　　处：《临床和实验医学杂志》2020年第3期225-229,共5页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金(编号:81650014)

摘　　要：目的探讨通用转录因子ⅡH亚基2(GTF2H2)在肝癌细胞中的核苷酸切除修复(NER)功能及对肝癌细胞增殖和凋亡的影响。方法使用siRNA构建GTF2H2敲低的肝癌细胞模型,采用细胞计数试剂盒(CCK8)实验检测肝癌细胞的增殖能力;流式细胞术检测肝癌细胞株的凋亡情况。定量实时聚合酶链式反应(q-RT-PCR)检测增殖细胞核抗原(PCNA)和凋亡蛋白半胱天冬酶(Caspase-3)的表达情况。进一步构建肝癌细胞的紫外线(UV)照射模型,分析GTF2H2敲低后UV照射引起的NER的标志物环丁烷嘧啶二聚体(CPD)和嘧啶(6-4)嘧啶酮光产物(6-4PP)的变化。结果GTF2H2敲低的肝癌细胞增殖能力相对明显增强增殖蛋白PCNA的表达相对增多。相反,凋亡能力相对减弱凋亡蛋白Caspase-3的表达相对降低。而GTF2H2敲低后,UV照射所致的NER标志物CPD和6-4PP含量相对增加。结论GTF2H2在肝癌细胞中发挥了NER功能,抑制了肝癌细胞的增殖,促进其凋亡,是肝癌发生的一个潜在抑制因子。Objective To investigate the proliferation and apoptosis effect of the Nucleotide Excision Repair(NER) function of general transcription factor Ⅱ H subunit 2(GTF2 H2) to liver cancer cells.Methods GTF2 H2 instantaneous knockdown liver cancer cells were constructed by using GTF2 H2-siRNAs.The proliferation ability of liver cancer cells was detected by CCK8 assay.The apoptosis of liver cancer cells was observed by flow cytometry.The expression of proliferating protein PCNA and apoptotic protein Caspase-3 was detected by real-time quantitative PCR.The UV irradiation model of liver cancer cells was further constructed to analyze the changes of NER markers CPD and 6-4 pp caused by Ultraviolet(UV) irradiation.Results The proliferation ability of liver cancer cells with transient knockdown of GTF2 H2 was significantly enhanced,and the expression of proliferating protein PCNA was increased.On the contrary,the apoptotic ability is relatively weakened,and the expression of apoptotic protein Caspase-3 was relatively reduced.Moreover,the contents of NER markers CPD and 6-4 pp in liver cancer cells with transient knockdown of GTF2 H2 after UV irradiation were relatively increased.Conclusion GTF2 H2 plays a NER role in liver cancer cells,and inhibits the proliferation of liver cancer cells and promotes their apoptosis.It might be a potential inhibiting factor for the occurrence of liver cancer.

关 键 词：GTF2H2 核苷酸切除修复 肝癌细胞 增殖 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]