艰难梭菌毒素A对K562细胞Rho GTPases及细胞骨架的影响  

Effects of Tcd A on Rho GTPases and the Cytoskeleton of Leukemia Cell Line K562

作　　者：赵龙[1] 陈彻 李明[1] 张豪[1] 刘蓓[1] 傅思武[3] 席亚明[1] ZHAO Long;CHEN Che;LI Ming;ZHANG Hao;LIU Bei;FU Si-Wu;XI Ya-Ming(Department of Hematology,Institute of Hematology,The First Hospital of Lanzhou University,Lanzhou 730000,Gansu Province,China;Teaching Center of Medical Basic Experiment,Gansu College of Traditional Chinese Medicine,Lanzhou 730000,Gansu Province,China;Medical Collage of Northwest University for Nationalities,China,Lanzhou 730000,Gansu Province,China)

机构地区：[1]兰州大学第一医院血液科,血液学研究所,甘肃兰州730000 [2]甘肃中医药大学临床医学院,甘肃兰州730000 [3]西北民族大学医学院,甘肃兰州730000

出　　处：《中国实验血液学杂志》2018年第5期1330-1335,共6页Journal of Experimental Hematology

基　　金：2014年兰州市科技计划项目(2014-1-37),2015年兰州大学第一医院院内基金(ldyyyn2015-09)。

摘　　要：目的:研究艰难梭菌毒素A(TcdA)对白血病K562细胞株Rho GTPase及细胞骨架的影响。方法:体外培养K562细胞经不同浓度的TcdA处理,采用四唑蓝比色试验(MTT)检测TcdA对K562细胞增殖的影响;RT-PCR法检测TcdA作用后细胞骨架调节蛋白相关基因cdc42、RhoA、Rac1 mRNA表达的变化;激光共聚焦显微镜观察TcdA作用48h后细胞微丝的变化。结果:TcdA能抑制K562细胞的增殖,作用48h后的抑制率(IR)为47.67%,TcdA可下调细胞骨架调节蛋白相关基因cdc42、RhoA、Rac1 mRNA表达,与对照组相比差异具有统计学意义(P <0.05)。激光共聚焦显微镜显示,TcdA处理后细胞内微丝含量下降。结论:艰难梭菌毒素A可抑制白血病细胞株K562增殖,其作用机制可能与Rho GTPase通路蛋白相关基因的表达下降,微丝形成受抑相关。Objective:To investigate the effect of clostridium difficile toxin A(TcdA)on the Rho GTPases and the cytoskeleton in K562 cells.Methods:K562 cells were cultured in vitro with different concentration of TcdA.The effect of TcdA proliferation of cells was detected by MTT method after the K562 cells were stimulated with TcdA for 24,48 and 72h;the expression of cdc42,RhoA,Rac1 mRNA was assessed by RT-PCR;the changes of the microtubule,the microfilament were observed by confocal laser scanning microscopy.Results:The proliferation of K562 cells was inhibited after exposure to TcdA for 24,48 and 72h,and the inhibitory rate was 47.67%in the treatment for 48 h.the cdc42,RhoA and Rac1 mRNA expressions in the experimental groups decreased after treated with TcdA(P<0.05),which positively correlated with concentration of TcdA.Also,the microfilament decreased,which was observed by confocal laser scanning microscopy.Conclusion:TcdA inhibites K562 cell proliferation and induces apoptosis,TcdA can change the cytoskeleton structure through the cytoskeletal protein genes cdc42 and RhoA,Rac1 mRNA expression,.It is related with cell microfilament content decreasing.

关 键 词：艰难梭菌毒素A K562细胞 RHO GTPASES RAC1 细胞骨架 

分 类 号：R733.7[医药卫生—肿瘤]