肿瘤靶向性细胞穿膜肽介导小分子干扰RNA对肝癌细胞的抑制作用  

Inhibitory effect of small interfering RNA mediated by tumor target cell penetrating peptides on in hepatoma cell line SMMC-7721

作　　者：何吉文[1,2] 李华[2] 向国安[1] 陈开运[1] 傅斌生[2] 杨扬[2] 陈规划[2] 

机构地区：[1]广东省第二人民医院普外一科,广州510317 [2]中山大学附属第三医院肝脏移植中心

出　　处：《中华实验外科杂志》2013年第10期2040-2043,共4页Chinese Journal of Experimental Surgery

基　　金：国家＂973＂计划资助项目（2009CB522404）;国家自然科学基金资助项目（30772135、81172038）;广东省科技计划资助项目（2009B030801078）

摘　　要：目的 观察以基质金属蛋白酶为靶点的肿瘤靶向性细胞穿膜肽介导小干扰RNA（siRNA）进入肝癌并对肝癌细胞SMMC-7721起一定抑制作用.方法 以体外培养的肝癌细胞SMMC-7721为研究对象,将细胞分为实验组和对照组,实验组加入siRNA质粒/穿膜肽复合物,对照组加入空质粒/穿膜肽复合物,培养48 h后收集两组细胞,流式细胞仪检测各组细胞的细胞周期,逆转录-聚合酶链反应（RT-PCR）检测各组细胞人端粒酶逆转录酶（hTERT）基因的表达水平,噻唑蓝（MTT）法检测SiRNA质粒/穿膜肽复合物对细胞增殖的抑制作用.结果 实验组与对照组比较G1期细胞所占比例增多（实验组75.14％、对照组62.55％）,G2期细胞比例减少（实验组11.39％、对照组12.14％）、S期细胞比例减少（实验组13.47％、对照组25.31％）,说明经穿膜肽质粒复合物处理后细胞被阻滞在G1期,实验组细胞hTERT mRNA的表达量约为未对照组的74％,即实验组mRNA表达水平下调约26％.肝癌细胞经穿膜肽复合物处理48 h后MTT法测细胞增殖抑制率为34.82％.结论 肿瘤靶向性细胞穿膜肽可介导siRNA进入肝癌细胞并可对肝癌细胞起一定的抑制作用.Objective To observe the inhibitory effect of small interfering RNA （siRNA） mediated by tumor target cell penetrating peptides （CPPS） targeting metalloproteinase on hepatoma cell line SMMC-7721.Methods SMMC-7721 cells were cultured,and divided into experimental group and control group.In experimental group,siRNA/CCPS complex was added,and in control group,silencer negative control/CCPS was added.The cells were harvested after culture for 48 h.Flow cytometry was use to examine the cell cycle.The human telomerase reverse transcriptase （hTERT） gene expression level was detected by using quantitative polymerase chain reaction （PCR）.Methyl thiazol tetrazolium （MTT） assay was used to meassure the inhibitory effect of siRNA/CCPS complex on SMMC-7721 cells.Results As compared with control group,the proportion of cells in G1 phase in experimental group was increased （75.14％ vs.62.55％）,while that in G2 phase in experimental group decreased （11.39％ vs.12.14％）,and that in the S phase also reduced （13.47％ vs.25.31％）.Treatment of CPPS plasmid complexes could arrest the cells in the G1 phase,and the expression quantity of hTERT mRNA in experimental group was 74％ of that in control group.After treatment with CPPS plasmid complexes for 48 h,the proliferation inhibition rate of SMMC-7721 cells was 34.82％.Conclusion The CCPS can be a transmitter to transfect siRNA into hepatoma cells and suppress hepatoma cells to some extent.

关 键 词：癌 肝细胞 穿膜肽 肿瘤靶向性 小干扰RNA 

分 类 号：R735.7[医药卫生—肿瘤]