Objective:To investigate the inhibitory effect of small interfering RNA(siRNA)on yes-associated protein(YAP) expression and adriamycin(ADM)chemosensitivity of hepatoma cell lines.Methods:The siRNA sequences were designed based on YAP gene and were transfected into four different kinds of hepatoma cells (PLC/PRF/5,Huh-7,MHCC 97H,MHCC 97L) through lipofectamineTM 2000.The inhibition result of YAP expression in hepatoma cells was measured by Western blotting and fluorescence microscope.The viability rate of hepatoma cells was measured by methyl thiazolyl tetrazolium(MTT)colori-metry after treating with different concentrations of ADM.Results:The siRNA expression vector to aim directly at YAP gene was constructed successfully.Western blotting revealed that transfection with siRNA could inhibit the expression of YAP pro-tein in hepatoma cell lines.The expression of YAP protein in hepatoma cell lines was specially and effectually inhibited by RNA interference(RNAi).MTT colorimetry detection result revealed that the viability rate of hepatoma cell lines significantly de-creased after transfection with YAP siRNA.Conclusions:Inhibiting the expression of YAP by siRNA can enhance the sensitivi-ty of hepatoma cell lines to ADM.%目的:研究用小片段干扰 RNA(small interfering RNA,siRNA)抑制肝癌细胞株中 YAP(yes-associated protein)表达后肝癌细胞株对阿霉素(adriamycin,ADM)的敏感性变化。方法:设计 YAP 基因的 siRNA 序列,用脂质体 LipofectamineTM 2000将 siRNA 转染至4株肝癌细胞株 PLC/PRF/5、Huh-7、MHCC 97H、MHCC 97L 中,在荧光显微镜下观察并采用蛋白免疫印迹(Western blotting)法检测肝癌细胞株中 YAP 的表达。将细胞以不同浓度的 ADM 处理后,采用四甲基偶氮唑蓝(methyl thialzolyl tetrazolium,MTT)比色法检测细胞的存活率。结果:成功构建针对 YAP 的 siRNA 表达载体。Western blotting 结果显示,肝癌细胞株经特异性 siRNA 转染后,YAP 蛋白的表达受到抑制,表明 RNA 干扰(RNA interference, RNAi)能特异、有效地抑制肝癌细胞株中 YAP 的表达。MTT 结果显示,转染 YAP siRNA 后肝癌细胞株的存活率受到明显抑制。结论:siRNA 抑制肝癌细胞株中 YAP 的表达可增强肝癌细胞株对 ADM 的敏感性。
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