siRNA下调AEG-1对大肠癌细胞体内成瘤影响的研究

Effect of AEG-1 silencing by siRNA on tumor formation in colorectal cancer cell line

目的探讨小干扰RNA(siRNA)下调星形胶质细胞升高基因-1(AEG-1)表达对大肠癌细胞HCT116体内成瘤的影响。方法采用siRNA下调的方法建立低表达AEG-1的结直肠癌稳定细胞株HCT116,采用实时荧光定量PCR(qRT-PCR)和蛋白免疫印迹法检测转染前后AEG-1的表达;用转染前后的细胞株进行裸鼠体内成瘤实验,4周后测量瘤体体积及质量。结果成功建立低表达AEG-1的结肠癌细胞模型HCT116,qRT-PCR和蛋白免疫印迹法证实采用siRNA下调AEG-1 mRNA和蛋白表达明显有效,4周后下调AEG-1表达的裸鼠瘤体体积[(94.53±27.31)mm^3 vs.(47.54±10.13)mm^3]和质量[(0.039±0.016)g vs.(0.013±0.002)g]明显减少(P均<0.05)。结论下调AEG-1的表达能有效抑制体内肿瘤的生长,AEG-1有望成为肿瘤基因治疗的靶点。

Objective To evaluate the effect of astrocyte elevated gene-1(AEG-1)silencing by siRNA on tumor formation in the colorectal cancer cell line HCT116.Methods A stable colorectal cancer cell line HCT116 with AEG-1 silencing was constructed.The expression levels of AEG-1 before and after transfection were quantitatively measured by qRT-PCR and Western blot.The tumor formation experiment was performed by using the cell lines before and after transfection.The tumor volume and weight were measured at 4 weeks.Results The colorectal cancer cell line HCT116 with AEG-1 silencing was successfully constructed.qRT-PCR and Western blot demonstrated that the expression levels of AEG-1 mRNA and protein were significantly down-regulated by siRNA.At 4 weeks after AEG-1 silencing,the tumor volume was significantly decreased from(94.53±27.31)mm^3 to(47.54±10.13)mm^3,and the tumor weight was remarkably reduced from(0.039±0.016)g to(0.013±0.002)g(both P<0.05).Conclusion AEG-1 silencing can effectively inhibit tumor growth,which becomes a promising target for tumor gene therapy.