TRIP13对B细胞淋巴瘤细胞增殖与凋亡的调控作用及机制  

Regulation of TRIP13 on Proliferation and Apoptosis of B-Cell Lymphoma Cells and Its Mechanism

作　　者：闫肃然 张青 刘星辰 原静静 张帆 周可树 YANSu-Ran;ZHANG Qing;LIU Xing-Chen;YUAN Jing-Jing;ZHANG Fan;ZHOU Ke-Shu(Department of Hematology,The Affiliated Cancer Hospital of Zhengzhou University,Henan Cancer Hospital,Zhengzhou 450008,Henan Province,China)

机构地区：[1]郑州大学附属肿瘤医院河南省肿瘤医院血液科,河南郑州450008

出　　处：《中国实验血液学杂志》2021年第5期1485-1492,共8页Journal of Experimental Hematology

基　　金：河南省自然科学基金(182300410347)。

摘　　要：目的:通过在细胞系Granta-519和JVM-2敲低/过表达TRIP13来探索TRIP13对B细胞淋巴瘤细胞增殖与凋亡的调控作用及可能的分子机制。方法:使用慢病毒转染技术构建敲低/过表达TRIP13的Granta-519细胞和JVM-2细胞及其对照组细胞,荧光显微镜判断慢病毒对细胞的转染效率,实时荧光定量PCR技术和蛋白免疫印迹法评估敲低/过表达效率,CCK-8法检测细胞增殖能力,Annexin V-APC单染法检测细胞凋亡率,PI染色法检测细胞周期,蛋白免疫印迹法检测P53、MDM4和BCL-2的表达水平。结果:敲低TRIP13后Granta-519和JVM-2细胞的增殖能力显著减弱而凋亡率显著升高,过表达TRIP13后细胞的增殖能力显著增强而凋亡率显著降低。敲低TRIP13后Granta-519细胞明显阻滞于G1期,JVM-2细胞明显阻滞于G1和G2/M期。在Granta-519中敲低TRIP13后BCL-2蛋白的表达降低,MDM4蛋白的表达升高;过表达TRIP13后MDM4蛋白的表达降低。在JVM-2细胞中过表达TRIP13后BCL-2蛋白的表达升高。结论:TRIP13促进B细胞淋巴瘤细胞的增殖,抑制其凋亡,并通过参与细胞周期的调控影响增殖和凋亡。B细胞淋巴瘤细胞中TRIP13促进BCL-2蛋白的表达,抑制MDM4蛋白的表达。Objective:To explore the regulatory effect of TRIP13 on the proliferation and apoptosis of B-cell lymphoma cells and its possible molecular mechanism by knocking down/overexpressing TRIP13 on the cell lines Granta-519 and JVM-2.Methods:Lentiviral transfection technology was used to construct Granta-519 and JVM-2 cells with knocked down or overexpressed TRIP13 and their control cells.The efficiency of transfection was determined by fluorescence microscopy.The efficiency of knockdown and overexpression was evaluated by real-time quantitative PCR and Western blot.The proliferation was detected by CCK-8 assay.The apoptosis was detected by the Annexin V-APC single staining.The cell cycle was detected by the PI staining.The expression levels of P53,MDM4,and BCL-2 were evaluated by Western blot.Results:After TRIP13 was knocked down,the proliferation ability of Granta-519 and JVM-2 cells was significantly reduced,and the apoptosis rate significantly increased.After TRIP13 was overexpressed,the proliferation ability of Granta-519 and JVM-2 cells was significantly enhanced,and the apoptosis was significantly reduced.After TRIP13 was knocked down,Granta-519 cells had obvious G1 phase arrest,and JVM-2 cells had obvious G1 and G2/M phase arrest.After TRIP13 was knocked down in Granta-519 cells,the expression of BCL-2 protein decreased,while MDM4 protein increased.After TRIP13 was overexpressed,the expression of MDM4 protein decreased.After TRIP13 was overexpressed in JVM-2 cells,the expression of BCL-2 protein increased.Conclusion:TRIP13 promotes the proliferation of B-cell lymphoma cells,inhibits their apoptosis,and affects their proliferation and apoptosis by participating in the regulation of the cell cycle.TRIP13 promotes the expression of BCL-2 proteins and inhibits the expression of MDM4 protein in B-cell lymphoma cells.

关 键 词：B细胞淋巴瘤 甲状腺激素受体相互作用因子13 增殖 凋亡 Granta-519 JVM-2 

分 类 号：R733.1[医药卫生—肿瘤]