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目的:观察癌胚抗原(CEA)特异性基因沉默对溶瘤腺病毒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(RNAi)技术,构建针对人食管癌EC9706细胞的CEAsiRNA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,并以空载体转染和未进行转染的EC9706细胞作对照,建立人食管癌细胞裸鼠皮下移植瘤模型,用H101进行瘤内注射。采用Real Time PCR和Western Blot方法检测移植瘤组织中CEA的表达,测量肿瘤体积。结果:降低CEA在mRNA和蛋白质水平的表达,对人食管癌EC9706细胞裸鼠皮下移植瘤成瘤时间和体积无影响,经H101瘤内注射治疗后,干扰组、空载体组和对照组肿瘤体积分别为(50.16±19.01)、(208.13±72.29)、(219.46±46.56)mm3,干扰组与对照组比较差异有统计学意义(P<0.05),而空载体组与对照组比较差异无统计学意义(P>0.05)。结论:抑制食管癌EC9706细胞中CEA的表达,可引起H101敏感性增加。
Objective: To observe the effect of carcinoembryonic antigen (CEA) -specific gene silencing on the effect of oncolytic adenovirus H101 on esophageal carcinoma EC9706 cells in order to explore the underlying factors that affect the sensitivity of H101. Methods: CEA siRNA vector targeting human esophageal carcinoma cell line EC9706 was constructed by RNA interference (RNAi) technology. After transfected with ECAs, the stable CEA gene silencing system was established in EC9706 cells and transfected with empty vector and untransfected EC9706 cells as control, establish human esophageal cancer nude mice subcutaneous xenograft model, with H101 for intratumoral injection. Real Time PCR and Western Blot were used to detect the expression of CEA in the tumor tissue and the tumor volume was measured. Results: Decreasing the expression of CEA at mRNA and protein level had no effect on the tumorigenic time and volume of subcutaneous xenografts of human esophageal carcinoma EC9706 cells. After intratumoral injection of H101, the tumor volume of the interfering group, empty vector group and control group (50.16 ± 19.01), (208.13 ± 72.29) and (219.46 ± 46.56) mm3, respectively. The difference between the interference group and the control group was statistically significant (P <0.05), while the difference between the empty vector group and the control group was not statistically significant Significance (P> 0.05). Conclusion: Inhibition of CEA expression in esophageal carcinoma EC9706 cells can cause an increase in H101 sensitivity.