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目的研究转染外源性CC10基因对肺腺癌A549细胞株的细胞周期及周期蛋白CyclinD1蛋白和mRNA表达影响。方法用脂质体法分别将外源性抑癌基因CC10转染到人肺腺癌细胞株A549中,8h、16h、24h、36h、48h后,分别用Western blot法检测转染CC10基因的A549细胞CC10蛋白表达,用流式细胞仪观察细胞周期的变化,用免疫细胞化学及RT-PCR检测转染CC10基因对周期蛋白CyclinD1蛋白、mRNA的影响。结果转染外源CC10基因对A549细胞生长具有抑制作用,细胞生长阻止于G0／G1,细胞周期S期+G2／M期比例下降。转染外源 CC10基因的A549细胞CC10蛋白表达增高,而CyclinD1的蛋白及mRNA表达明显降低。结论转染外源CC10基因对肺癌细胞G0／G1周期的抑制作用可能与CyclinD1基因表达下调有关。
Objective To investigate the effect of transfection of exogenous CC10 gene on cell cycle and the expression of cyclinD1 protein and mRNA in lung adenocarcinoma A549 cell line. METHODS: The exogenous tumor suppressor gene CC10 was transfected into human lung adenocarcinoma cell line A549 by liposome method. After 8h, 16h, 24h, 36h, and 48h, Western blot was used to detect A549 transfected with CC10 gene, respectively. The expression of CC10 protein was detected by flow cytometry. The effect of transfected CC10 gene on cyclinD1 protein and mRNA was detected by immunocytochemistry and RT-PCR. Results The transfection of exogenous CC10 gene inhibited the growth of A549 cells. The growth of cells was inhibited by G0/G1, and the proportion of cell cycle S+G2/M phase decreased. The expression of CC10 protein in A549 cells transfected with exogenous CC10 gene was increased, while the protein and mRNA expression of CyclinD1 was significantly decreased. Conclusion The inhibitory effect of transfection of exogenous CC10 gene on the G0/G1 cycle of lung cancer cells may be related to the down-regulation of CyclinD1 gene expression.