CEA基因沉默对基因工程腺病毒(H101)杀伤食管癌EC9706细胞的影响

目的: 研究癌胚抗原(CEA)特异性基因沉默对基因工程腺病毒(H101)杀伤食管癌EC9706细胞作用的影响,以探讨影响H101敏感性的内在因素。 方法: 利用RNA干扰技术,构建针对人食管癌EC9706细胞的CEA siRNA载体,通过基因转染,在EC9706细胞中建立稳定的CEA基因沉默体系(干扰1组及干扰2组),并以空载体转染和未进行转染的EC9706细胞作对照,用H101在体外进行细胞毒性实验。 结果: 在此实验任何病毒浓度下,干扰2组与空载体组和对照组比较,生存率明显下降,有统计学意义(P<0.05);而干扰1组在低病毒浓度(≤1vp/cell)下,与空载体组和对照组比较,生存率下降,有统计学意义(P<0.05),当病毒浓度>1vp/cell时,无统计学意义(P>0.05)。 结论: 提示CEA在H1001感染并杀伤肿瘤细胞过程中可能起一定作用,为进一步从基因水平探讨CEA的作用机制和提高溶瘤腺病毒的治疗效果打下基础。

The Killing Effect of Genetically Engineered Adenovirus(H101)in Conjunction with CEA Gene Silencing in EC9706 Esophageal Carcinoma Cells

Objective: To study the killing effects of genetically engineered adenovirus (H101) in conjunction withCEA genespecific silencing on EC9706 esophageal carcinoma cells and to explore the internal factors influencing H101sensitivity. Methods: We constructed siRNA expression vectors with multiple sequences targeting CEA and transfectedthem into EC9706 cells. Clones with stable CEA gene silencing were selected (interference group 1 and interference group2). Cells with empty vector and untransfected cells served as the controls for cytotoxicity experiments that were then per-formed with H101. Results: There was an evident decrease in survival rate in interference group 2 compared with thecontrol at any given virus density(P<0.05). Compared with the control group, interference group 1 exhibited decreased sur-vival when the virus density was equal to or less than 1 vp/cell (P<0.05); when the virus density exceeded 1 vp/cell, nosignificant difference was found in cell survival between the two groups (P>0.05). Conclusion: Cell survival was signifi-cantly different between the interference groups and the control group. CEA gene silencing in EC9706 cells can decreaseCEA expression and increase the death rate of cancer cells.