敲低转录共激活子对胶质瘤细胞生物学特性的影响  

Effect of knocking down the transcriptional coactivator on the cell biological characteristics of glioma

作　　者：栗伟杰 王广秀[1] 于福华[1] 张安玲[1] 康春生[1] 韩磊[1] 浦佩玉[1] 贾志凡[1] 

机构地区：[1]天津医科大学总医院神经病学研究所神经肿瘤室,300052

出　　处：《中华神经外科杂志》2015年第6期620-624,共5页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金（30872985、81101915）

摘　　要：目的 研究敲低带PDZ结合域的转录共激活子（transcriptional coactivator with PDZ-binding motif,TAZ）表达后对胶质瘤细胞株细胞生物学特性的影响.方法 采用TAZ小干扰RNA（TAZsiRNA）敲低LN229和SNB19细胞株中TAZ表达,用实时定量聚合酶链式反应（real time-PCR） 及蛋白免疫印迹法（Western blot）分别鉴定转染后TAZ敲除效果,噻唑兰比色法检测两种细胞株的增殖能力变化;流式细胞术检测细胞凋亡的改变;划痕实验和Transwell实验检测细胞侵袭能力的变化;Western blot法检测凋亡、增殖及侵袭等相关蛋白的表达变化.结果 转染TAZ siRNA可显著敲低LN229和SNB19细胞株中TAZ mRNA及蛋白表达水平;两种细胞株中TAZ被抑制后,细胞增殖率均下降且呈逐渐下降趋势,而细胞凋亡率分别增高至（12.05±0.65）％和（8.02 ±0.66）％（LN229：F=414.2,P=0.000;SNB19：F=156.8,P=0.000）;两细胞株TAZ siRNA组细胞穿膜细胞数与对照组和无义序列组相比明显降低,划痕间隙融合能力也显著降低,差异均有统计学意义（P＜0.05）;Western blot法检测显示转染后细胞增殖核抗原（Kiel 67 antigen,Ki-67）、B淋巴细胞瘤2（B-cell lymphoma-2,Bcl-2）、基质金属蛋白酶9（matrix metallopeptidase 9,MMP-9）等增殖、凋亡及侵袭相关蛋白表达明显下调.结论 敲低TAZ可抑制胶质瘤细胞株LN229和SNB19增殖和侵袭能力,提示TAZ可能成为治疗胶质瘤的新靶点.Objective To study the effect of knocking down the transcriptional coactivator with PDZ-binding motif （TAZ） expression on the biological characteristics of glioma cell lines.Methods The TAZ small interfering RNA （TAZ siRNA） was used to knock down TAZ expression in LN229 and SNB19 cells.Real-time quantitative polymerase chain reaction （RealTime-PCR） and Western blot were used to identify the effect of TAZ knockout after transfection.Thiazole blue （MTT） colorimetric assay was used to detect the changes of the proliferation ability of 2 kind cells.Flow cytometry was used to detect the changes of apoptosis.Scratch experiment and Transwell experiment were used to detect the changes of cell invasion ability.Western blot was used detect the expression changes of related proteins including apoptosis,proliferation and invasion,etc.Results The transfection of TAZ siRNA significantly knocked down the expression levels of TAZ mRNA and protein in LN229 and SNB19 cells ; after TAZ was inhibited in the 2 kinds of cells,the cell proliferation rates were decreased and showed a gradual downward trend,while the apoptosis rates were increased to 12.05% ±0.65% and 8.02% ±0.66% （LN229：F =414.2,P =0.000;SNB19：F =156.8,P =0.000）.Compared with the control and nonsense sequence groups,the cell numbers of the cell-penetrating membrane per field were decreased significantly in the TAZ siRNA group in both cell lines.The fusion capability of scratch gap was also decreased significantly,and there was significant difference （P =0.05）.Western blot for the proliferations of cellular proliferative nuclear antigen （Kiel67 antigen,Ki-67） B-cell lymphoma-2 （Bcl-2） and matrix metallopeptidase 9 （MMP-9）,apoptosis and invasion-related protein expression were down-regulated significantly.Conclusion Knocking down TAZ inhibits proliferations and invasive abilities of LN229 and SNB19 of the glioma cell lines,suggesting TAZ may become a new target for the treatment of gliomas.

关 键 词：神经胶质瘤 细胞增殖 肿瘤侵润 凋亡 带PDZ结合域的转录共激活子 

分 类 号：R739.41[医药卫生—肿瘤]