靶向survivin利用RNA干扰技术影响云南个旧肺鳞癌细胞株YTMLC凋亡的初步研究
RNA Interference-mediated Significant Enhancement of Apoptosis in Yunnan Gejiu Lung Squamous Carcinoma Cells(YTMLC) by siRNA Expression Vector Targeting Survivin Gene
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摘要: 目的检测云南个旧肺鳞癌细胞株YTMLC细胞内源凋亡抑制因子survivin基因的表达以及促进YTMLC细胞凋亡的情况。方法构建和转染重组质粒pshRNA-survivin-1和pshRNA-survivin-2至YTMLC细胞株,通过免疫荧光和半定量RT-PCR检测survivin蛋白表达及mRNA转录水平的变化,并通过流式细胞仪使用PI-AnnexinV双染法检测YTMLC细胞株的凋亡。结果构建的2种重组质粒pshRNA-survivin-1和pshRNA-survivin-2均能明显抑制survivin基因的表达;应用免疫荧光检测survivin基因的表达,转染重组质粒pshRNA-survivin-1和pshRNA-survivin-2的实验组survivin荧光强度明显低于转染空载体pGE-1和pshRNA-negative非特异对照质粒;通过半定量RT-PCR检测到sur-vivin基因mRNA转录明显减少,抑制率为80.60%和70.09%;通过PI-AnnexinV双染法检测YTMLC细胞的凋亡分别达(33.42±2.42)%(P<0.01)和(20.09±1.32)%(P<0.01)。结论重组...Abstract: Objective To apply the small interfering RNAs targeting survivin to inhibit expression of endogenous survivin gene in Yunnan Gejiu lung squamous carcinoma cells(YTMLC) and increase the apoptosis of Yunnan Gejiu lung squamous carcinoma cells(YTMLC). Methods The recombinant plasmid pshRNA-survivin-1 and pshRNA-survivin-2 were constructed and transfected into YTMLC cell line.The expression level of survivin was determined by immunofluorescence staining and the transcription of survivin gene was detected by semi...
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