RNA干扰FABP5基因对人肝癌HepG2细胞裸鼠移植瘤生长的影响

目的:研究脂肪酸结合蛋白5(FABP5)基因沉默的慢病毒载体对人肝癌Hep G2细胞成瘤效应的影响。方法:采用RNA干扰技术构建重组逆转录慢病毒载体。Hep G2细胞分为3组:实验组以FABP5基因沉默慢病毒颗粒(LV-shRNA-FABP5)感染Hep G2细胞;阴性对照组以空载体慢病毒颗粒(LV-shRNA-NC)感染Hep G2细胞;空白对照组不做任何处理。将裸鼠随机分为3组,接种肿瘤细胞后观察裸鼠成瘤情况。4周后测量肿瘤的体积和重量,绘制移植瘤生长曲线。Real-time PCR、Western blot及免疫组织化学法检测裸鼠移植瘤中FABP5的表达。结果:LV-shRNA-FABP5可以降低Hep G2细胞FABP5的表达。3组裸鼠接种癌细胞后均有肿瘤形成。与空白对照组和阴性对照组相比,实验组肿瘤生长速度明显减慢,且体积及重量明显减小(P0.05);实验组裸鼠肝癌移植瘤组织的FABP5 mRNA和蛋白表达水平相比空白对照组和阴性对照组表达明显下降(P0.05)。结论:沉默FABP5基因表达能有效抑制人肝癌裸鼠移植瘤的生长。FABP5可能成为肝癌基因治疗的一个有效靶点。

Effects of RNA interference of FABP5 on subcutaneous tumor of human hepatocellular carcinoma cell line HepG 2 transplanted in nude mice

AIM: To investigate the effect of recombinant lentiviral vector for RNA interference (RNAi) on the expression of fatty acid-binding protein 5 (FABP5) gene in hepatocellular carcinoma HepG2 cells and tumor formation in nude mice.METHODS: RNAi lentiviral vector was used in the experiment. Human hepatocellular carcinoma HepG2 cells were divided into 3 groups:the HepG2 cells in experimental group were transfected with the recombinant lentivirirus vector LV-shRNA-FABP5, the cells in negative control group were transfected with a control lentiviral vector LV-shRNA-NC, and the cells in normal control group were without any treatment. The nude mice were randomly divided into 3 groups. The growth of the transplanted tumor cells in the nude mice was observed. The tumor growth curve, volume and weight were determined 4 weeks after the cell inoculation. The expression of FABP5 was detected by real-time PCR, Western blot and immunohistochemical staining.RESULTS: Transfection of the lentiviral vector FABP5-shRNA obviously reduced FABP5 expression in the HepG2 cells. Tumor formation was all positive in the 3 groups of the nude mice inoculated with the tumor cells. Compared with normal control group and negative control group, the tumor growth slowed significantly in experimental group with smaller volume and weight. FABP5 expression in the transplanted tumor tissues was significantly down-regulated at mRNA and protein levels in experimental group as compared with normal control group and negative control group.CONCLUSION: RNAi-induced down-regulation of FABP5 effectively inhibits the growth of transplanted hepatocellular carcinoma, suggesting that FABP5 gene may be an effective target for gene therapy in treating liver cancer.