敲减NLRC3表达对人正常支气管上皮BEAS-2B细胞生长的影响  

Effect of NLRC3 expression knock-down on growth of normal human bronchial epithelial BEAS-2B cells

作　　者：黄铀新[1] 孟祥磊 张敏鸿[3] 罗耀玲[3] 李杰[2] HUANG You-xin;MENG Xiang-lei;ZHANG MIN-hong;LUO YAO-ling;LI Jie(Department of Nuclear Medicine,2Department of Resperatory Medicine,3Clinical Research Center,The First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,China)

机构地区：[1]赣南医学院第一附属医院核医学科,江西赣州341000 [2]赣南医学院第一附属医院呼吸内科,江西赣州341000 [3]赣南医学院第一附属医院临床科研中心,江西赣州341000

出　　处：《中国病理生理杂志》2020年第6期985-990,共6页Chinese Journal of Pathophysiology

基　　金：江西省教育厅科技研究项目(No.GJJ150946);赣南医学院科研课题(No.YB201503)。

摘　　要：目的:探讨敲减含胱天蛋白酶募集结构域的NOD样受体家族蛋白3(NLRC3)表达对人正常支气管上皮细胞株BEAS-2B活力和凋亡的影响及机制。方法:使用Lipofectamine 2000转染试剂将NLRC3基因的小干扰RNA(siRNA)片段转染至BEAS-2B细胞;RT-qPCR筛选干扰片段;MTT法检测细胞活力;JC-1检测细胞线粒体膜电位变化;annexin V-FITC/PI凋亡试剂盒检测细胞凋亡率;Western blot检测Bcl-2和Bax的表达水平。结果:NLRC3基因的干扰片段3(siNLRC3-3)干扰效果最佳(P<0. 01)。在BEAS-2B细胞中敲减NLRC3可显著增强细胞活力(P<0. 01)。敲减NLRC3表达可显著升高线粒体膜电位,使细胞凋亡率显著下降(P<0. 05)。敲减NLRC3表达使细胞中Bcl-2蛋白表达水平显著上调,而Bax蛋白表达水平显著下调(P<0. 01)。结论:敲减NLRC3基因可通过上调Bcl-2蛋白表达、下调Bax蛋白表达,增强BEAS-2B细胞活力并抑制其凋亡。AIM:To investigate the effect of NOD-like receptor family caspase recruitment domain containing 3(NLRC3)expression knock-down on the viability and apoptosis of normal human bronchial epithelial BEAS-2B cells and its mechanism.METHODS:The small interfering RNA(siRNA)fragments of NLRC3 gene were transfected into BEAS-2B cells using Lipofectamine 2000 transfection reagent to knock down the NLRC3 expression.The interference fragment was screened by RT-qPCR.The cell viability was measured by MTT assay.The mitochondrial membrane potential was de⁃tected by JC-1 staining.The apoptotic rate was analyzed by flow cytometry with annexin V-FITC/PI staining.The protein expression levels of Bcl-2 and Bax were determined by Western blot.RESULTS:The interference segment 3 of NLRC3 gene(siNLRC3-3)displayed the best interference effect on NLRC3 expression in BEAS-2B cells(P<0.01).Knock-down of NLRC3 expression in BEAS-2B cells enhanced the cell viability(P<0.01).Knock-down of NLRC3 increased the mito⁃chondrial membrane potential,and decreased the apoptotic rate(P<0.05).Moreover,knock-down of NLRC3 significant⁃ly up-regulated Bcl-2 protein expression and significantly down-regulated Bax protein expression(P<0.01).CONCLU⁃SION:Knock-down of NLRC3 expression enhances the viability and inhibits the apoptosis of BEAS-2B cells,which may be related to increase in the expression of Bcl-2 protein and decrease in the expression of Bax protein.

关 键 词：肺癌 NLRC3蛋白 细胞活力 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤] R730.23[医药卫生—临床医学]