过表达核苷二磷酸X水解酶21(NUDT21)通过阻断P53/CDK2/Rb通路抑制HCT-116结肠癌细胞的增殖  

Over-expression of NUDT21 inhibits the proliferation of HCT-116 colon cancer cells via blocking P53/CDK2/Rb pathway

作　　者：罗珍 豆梁丁 王蕾 刘榕 罗光平 林默 邓子峰 沈颖 傅芷宁 彭书海 张永兴[1] LUO Zhen;DOU Liangding;WANG Lei;LIU Rong;LUO Guangping;Lin Mo;DENG Zifeng;SHEN Ying;FU Zhining;PENG Shuhai;ZHANG Yongxing(State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics,School of Public Health,Xiamen University,Xiamen 361102;Department of Stomatology,Xiamen Medical College,Xiamen 361023,China)

机构地区：[1]厦门大学公共卫生学院分子疫苗学和分子诊断学国家重点实验室,福建厦门361102 [2]厦门医学院口腔医学系,福建厦门361023

出　　处：《细胞与分子免疫学杂志》2022年第6期507-512,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：中央高校基本科研业务专项资助基金(20720192016);厦门大学公共卫生学院本科生创新实践平台(NO.202004,NO.202005)。

摘　　要：目的探讨过表达核苷二磷酸X水解酶21(NUDT21)对HCT-116结肠癌细胞增殖的影响及可能的机制。方法Gibson组装法构建NUDT21过表达质粒。克隆形成实验、CCK-8法检测细胞增殖活性,流式细胞术检测细胞周期变化,Western blot法检测P53、细胞周期蛋白依赖激酶2(CDK2)的蛋白水平,以及第780位丝氨酸磷酸化的视网膜母细胞瘤蛋白(p-Rb-Ser780)和p-Rb-Ser608的蛋白水平。结果测序结果提示NUDT21过表达载体构建成功,HCT-116细胞转染NUDT21过表达质粒后,其mRNA和蛋白水平表达量明显升高。过表达NUDT21抑制结肠癌HCT-116细胞增殖,细胞周期阻滞于G0/G1期;过表达NUDT21后,CDK2、p-Rb-Ser608与p-Rb-Ser780蛋白表达量降低,P53蛋白水平增高。结论过表达NUDT21通过阻断P53/CDK2/Rb信号通路抑制结肠癌HCT-116细胞的增殖。Objective To investigate the effect of over-expression of nudix hydrolase 21(NUDT21)on the proliferation of colon cancer HCT-116 cells and its mechanism.Methods The NUDT21 over-expression plasmid was constructed by Gibson assembly.The colony formation assay and CCK-8 assay were used to detect cell proliferation.The cell cycle of HCT-116 cells was detected by flow cytometry.Western blot was performed to detect the expressions of P53,cyclin-dependent kinase 2(CDK2),phosphorylated retinoblastoma protein at serine 780(p-Rb-Ser780),and p-Rb-Ser608.Results The sequencing results showed that the NUDT21 over-expression plasmid was successfully constructed.After the NUDT21 over-expression plasmid was transfected into HCT-116 cells,the expressions of NUDT21 mRNA and protein in the cells were significantly increased.The over-expression of NUDT21 inhibited the proliferation of HCT-116 cells and arrested the cell cycle in G0/G1 phase.The expressions of CDK2,p-Rb-Ser608,and p-Rb-Ser780 proteins decreased while the expression of P53 protein increased.Conclusion Over-expression of NUDT21 inhibits the proliferation of HCT-116 cells by blocking P53/CDK2/Rb signal pathway.

关 键 词：核苷二磷酸X水解酶21(NUDT21) HCT-116细胞 细胞增殖 细胞周期 

分 类 号：Q279[生物学—细胞生物学] R735.3[医药卫生—肿瘤] R291-33[医药卫生—临床医学] R34Q253