外源CC10基因对肺腺癌A549细胞细胞周期蛋白D1 mRNA表达的影响

Effects of exogenous CC10 gene on CyclinD_1 mRNA expressions in the A549 lung cancer cells

目的:研究外源性CC10基因对肺腺癌A549细胞细胞周期蛋白D1(CyclinD1)及其mRNA表达的影响。方法:用脂质体法将抑癌基因CC10转染到人肺腺癌细胞A549中8、16、24、32、48h后,用Western blot法观察CC10蛋白表达,观察细胞生长细胞克隆形成,用流式细胞仪观察细胞周期变化,用RT-PCR检测周期蛋白CyclinD1 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期,CyclinD1 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 effects of exogenous CC10 gene on CyclinD1 mRNA expression in A549 cell line. Methods CC10 gene was transfected into A549 cell by medium with liposome for 8, 16, 24, 32 and 48 h. The change of CC10 protein expression in A549 cell line were observed by Western blot. The growth inhibitory rate was detected by MTT method. Flow cyometry analysis and AnnexinV-PI staining were used to determine the changes of cell cycle progression and apoptosis. CyclinD1 mRNA expression in A549 cell was detected by RT-PCR. Results Exogenous CC10 gene inhibited the A549 cells growing, and the growth inhibitory rate were 24.7%, 33.1%, 44.3%, 61.7% and 74.2%, respectively. CyclinD1 protein and mRNA expression were lower in A549 cells transfected by CC10 gene than that in normal A549 cells. Conclusion It is found that CyclinD1 mRNA expressions decrease in A549 cells transfected by CC10 gene, which may be correlated with cell cycle progression inhibiting in G0/G1 phase.