c35在肝癌中的表达与靶向干预研究.docVIP

C35在肝癌中的表达与靶向干预研究 　　【摘要】 目的：?^察探讨C35在肝癌中的表达与靶向干预，为临床治疗提供依据。方法：设计两对C35编码区的siRNA并合成，构建到转录载体pTZU6+1上，形成重组质粒siRNA-C35，在脂质体介导下转染肝癌细胞株，RT-PCR分析RNA干扰后mRNA的变化，Western blot检测表达蛋白的变化。结果：扩增的目的基因条带亮度存在明显差异，与对照组相比，转染质粒pTZU6+1相对表达率为95.3%、siRNA1组相对表达率为40.0%，siRNA2组的相对表达率为28.4%。构建的重组质粒对C35mRNA的表达有抑制作用。只转染脂质体组和pTZU6+1的C35蛋白杂交带强于siRNA1和siRNA2，证实重组质粒可以抑制C35基因的表达，以对照组的蛋白带为标准，其图像分析可知，转染质粒pTZU6+1相对表达率为94.9%、siRNA1组相对表达率为29.6%，siRNA2组的相对表达率为32.2%。结论：通过设计C35基因敲除的siRNA可有效抑制C35基因的表达，肝癌细胞生长速度及侵袭性均发生改变，但C35基因在肝癌细胞株中及肝癌组织中低表达，不可作为肝癌的靶点干预基因。 　　【关键词】 C35； 肝癌； 表达； 靶向干预 　　Study of C35 Expression in Hepatocellular Carcinoma and Targeted Intervention/ZHAN Yuan-jing，HU Zhong-wei，GUO Jia-wei，et al.//Medical Innovation of China，2017，14（16）：025-028 　　【Abstract】 Objective：To investigate the expression of C35 in hepatocellular carcinoma（HCC），and provide the basis for clinical treatment.Method：Designed and compounded two groups C35 coding rigion siRNA，transcription vector pTZU6+1 to construct recombinant plasmid siRNA-C35，liposome mediated transfection for hepatocellular carcinoma cell lines，the change of mRNA protein expression after RNA interference by RT-PCR，the change of protein detected by Western blot.Result：There was significant difference in the brightness of the target gene.Compared with control group，the relative expression rates of the transfected plasmid pTZU6+1，siRNA1 and siRNA2 were respectively 95.3%，40.0% and 28.4% respectively.The constructed recombinant plasmid could inhibit the expression of C35mRNA.The transfection of C35 protein only with liposome and pTZU6+1 were stronger than that of siRNA1 and siRNA2，which further confirmed that the recombinant plasmid had inhibitory effect on C35 expression.By the standard of the the control groups’ protein band，the analysis of the image system showed the protein expression of the transfected plasmid pTZU6+1，siRNA1 and siRNA2 group were respectively 94.9%，29.6% and 32.2% respectively.Conclusion：Through the design of the C35 knockout siRNA can e