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目的:研究肝激酶B1(LKB1)基因对大肠癌细胞增殖的影响。方法:用脂质体转染法将LKB1基因表达质粒(pReceiver-LKB1)瞬时转染SW1116及SW480细胞,半定量RT-PCR及Western blot方法检测LKB1 mRNA和蛋白水平的表达;CCK8法检测SW1116及转染重组质粒的细胞体外增殖变化;半定量RT-PCR和Western blot方法检测胞内LKB1对细胞周期蛋白cyclin D1的影响。结果:转染LKB1基因后,质粒转染组LKB1表达较空质粒组及空白对照组明显升高;质粒转染组在转染后48、72、96 h时,细胞活性较空质粒组及空白对照组明显降低(P<0.01);与对照组相比,质粒转染组的细胞内cyclin D1 mRNA和蛋白的表达明显降低。结论:LKB1可抑制大肠癌细胞的体外增殖能力,并且这一抑制效应可能通过下调cyclin D1产生。
Objective: To study the effect of hepatic kinase B1 (LKB1) gene on the proliferation of colorectal cancer cells. METHODS: SW1116 and SW480 cells were transiently transfected with the LKB1 gene expression plasmid (pReceiver-LKB1) by liposome transfection. Semi-quantitative RT-PCR and Western blot were used to detect the expression of LKB1 mRNA and protein. CCK8 method was used to detect SW1116 and SW480 cells. The proliferation of cells transfected with recombinant plasmids was detected in vitro. Semiquantitative RT-PCR and Western blot were used to detect the effect of intracellular LKB1 on cyclin D1. RESULTS: After transfected with LKB1 gene, the expression of LKB1 in plasmid transfection group was significantly higher than that of empty plasmid group and blank control group. At 48, 72, and 96 hours after transfection, the activity of LKB1 in plasmid transfection group was higher than that of empty plasmid group and blank. The control group significantly decreased (P <0.01); compared with the control group, the expression of cyclin D1 mRNA and protein in the plasmid transfection group was significantly reduced. Conclusion: LKB1 can inhibit the proliferation of colorectal cancer cells in vitro, and this inhibitory effect may be through down-regulation of cyclin D1 production.