沉默肝癌衍生生长因子对大肠癌细胞增殖的抑制

Effects of specific HDGF siRNA on proliferation of human colon carcinoma cells

目的:研究沉默肝癌衍生生长因子(hepatoma-derived growth factor,HDGF)对人大肠癌lovo细胞增殖的影响.方法:将3条HDGF的特异性小干扰RNA(HDGF-siRNA)通过脂质体介导瞬时转染入大肠癌lovo细胞,RT-PCR及Western blot法分别检测HDGFmRNA和蛋白质表达受抑程度,并筛选出1条沉默效率最高的siRNA用于后续试验;MTT法观察HDGF-siRNA对大肠癌细胞增殖生长的影响;Westernblot法检测增殖细胞核抗原(PCNA)及ERK通路蛋白在转染前后大肠癌lovo细胞中表达含量的变化.结果:HDGF-siRNA转染能有效抑制大肠癌lovo细胞中HDGF的表达;与空白及阴性对照组相比,RNA干扰组大肠癌lovo细胞增殖率明显降低(t=4.432,4.263,均P<0.01)、PCNA和p-ERK1/2蛋白均明显下调(t=14.89,11.92,均P<0.01).结论:HDGF-siRNA不仅有效阻断HDGF在大肠癌lovo细胞的表达,而且明显抑制大肠癌lovo细胞的增殖,其作用机制可能与下调PCNA,抑制ERK1/2蛋白活化有关.

AIM： To study the effect of specific HDGF- siRNA on proliferation of human colon carcinoma lovo cells. METHODS： Three pairs of specific HDGF-siRNA were transiently transfected into lovo cells by liposome and then the expression of HDGF was detected by RT-PCR and Western blot. The most efficient siRNA was selected for following experiments. Cell proliferation was examined by MTT assay, Finally we also detected the expression of proliferating cell nuclear antigen （PCNA） by RT-PCR and Western blot. RESULTS： After the specific siRNA was transfected into colon carcinoma lovo cells, the expression level of HGDF showed a significantly decrease which was detected by RT-PCR and Western blot. The cell survival rate was signifi-cantly lower in interference group than in blank control group or in negative control group （t = 4.432, 4.263, both P 〈 0.01）, and the expression levels of PCNA and ERK1/2 protein were lower in interference group than in the other two groups （t = 14.89, 11.92, both P 〈 0.01）. CONCLUSION： The results show that the specific HDGF-siRNA can effectively silence the expression of HDGF, and HDGF-siRNA significantly inhibit proliferation of colon carcinoma cell line lovo cells, accompanied by down-regu- lation of PCNA and p-ERK1/2.