PLCε调节肾细胞癌血管生成的作用机制  

Action Mechanism of Phospholipase C Epsilon in Regulating Angiogenesis of Renal Cell Carcinoma

作　　者：王春远[1] 罗春丽[1] 杨淑哲[1] 潘翠翠[1] 颜令[1] 张彦懿[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国生物制品学杂志》2011年第4期443-448,共6页Chinese Journal of Biologicals

摘　　要：目的探讨血管内皮生长因子(Vascular endothelial growth factor,VEGF)在PLCε-NF-κB信号通路中影响肿瘤血管生成的作用机制。方法将PLCε-shRNA表达质粒pGenesil-PLCε转染人肾透明细胞癌786-0细胞株,沉默磷脂酶Cε(Phos-pholipase C epsilon,PLCε)基因的表达,采用RT-PCR及Western blot检测转染细胞中PLCε、NF-κB和VEGF基因mRNA及蛋白水平表达的变化;应用NF-κB特异性抑制剂BAY11-7082处理细胞后,采用MTT法检测BAY11-7082对786-0细胞增殖的抑制作用,RT-PCR和Westernblot检测VEGF基因mRNA和蛋白水平表达的变化。结果重组质粒pGenesil-PLCε可明显抑制PLCε基因mRNA和蛋白水平的表达,抑制率分别为71.43%和50.01%,并明显下调NF-κB和VEGF基因mRNA和蛋白水平的表达;BAY11-7082可明显抑制786-0细胞增殖,且呈剂量和时间依赖性;应用BAY11-7082后,VEGF基因mRNA和蛋白的表达均被明显抑制。结论 PLCε可能通过抑制NF-κB基因的表达,从而抑制NF-κB依赖性基因VEGF的表达,进而影响肾细胞癌血管生成。Objective To investigate the mechanism of vascular endothelial growth factor（VEGF） in influencing the angiogenesis of tumors through phospholipase C epsilon（PLCε）-NF-κB signal pathway.Methods Plasmid pGenesil-PLC for expression of PLC ε-shRNA was transfected to human renal cell carcinoma 786-0 cell strain to silence the expression of PLCε gene.The expressions of PLC ε,NF-κB and VEGF in transfected cells at mRNA level were determined by RT-PCR,while those at protein level by Western blot.The cells were treated with BAY11-7082,a specific inhibitor of NF-κB,and determined for inhibiting effect on cell proliferation by MTT method,then for expressions of VEGF mRNA and protein by RT-PCR and Western blot respectively.Results Recombinant plasmid pGenesil-PLCε inhibited the expressions of PLCε at mRNA and protein levels significantly,of which the inhibiting rates were 71.43% and 50.01% respectively.The plasmid also down-regulated the expressions of NF-κB and VEGF mRNAs and proteins significantly.BAY11-7082 showed significantly dose-and time-dependent inhibitory effect on the proliferation of 786-0 cells.Both the expressions of VEGF mRNA and protein in the cells after treatment with BAY11-7082 were inhibited significantly.Conclusion PLCε might inhibited the expression of NF-κB-dependent VEGF gene by inhibiting the expression of NF-κB gene and influence the angiogenesis of renal cell carcinoma.

关 键 词：癌 肾细胞 磷脂酶C RNA干扰 基因沉默 血管内皮生长因子类 

分 类 号：R737.11[医药卫生—肿瘤]