目的:探究高表达和干扰PSMA7对A549细胞周期以及RB途径中Cyclin D1、CDK4、P16、Rb蛋白水平的影响。方法:将pcDNA3.1-PSMA7高表达和pGPU6/Hygro-PSMA7-265干扰载体分别瞬时转染A549细胞,然后用流式细胞仪检测PSMA7对细胞周期的影响,再通过Western blot实验检测Cyclin D1、CDK4、P16、Rb的蛋白水平。结果:和对照组相比,PSMA7高表达时周期时相分布无明显差异,但Cyclin D1、CDK4的蛋白表达水平明显降低,P16、Rb的表达明显增高;和对照组相比,干扰PSMA7后细胞的G0/G1、G2/M期比例均降低,而S期的值增高,均具有差异,而Cyclin D1、CDK4的蛋白表达水平明显增加,P16、Rb的表达明显降低,以上结果与对照组相比差异均有统计学意义。结论:PSMA7在A549肺腺癌细胞中能够影响RB途径中Cyclin D1、CDK4、P16、Rb的蛋白水平的表达,干扰PSMA7使A549细胞较早进入S期。
Objective: To investigate the effect of upregulated and downregulated PSMA7 on the cell cycle and Cyclin D1,CDK4,P16,Rb of RB pathway in A549 cells. Methods: Transfected upregulated pcDNA3. 1-PSMA7 vecter and downregulated pGPU6/Hygro-PSMA7-265 vecter into A549 cells,and then tested the effect of PSMA7 on the cell cycle of A549 cells by flow cytometry,and detected the protein level of Cyclin D1,CDK4,P16,Rb by Western blot. Results: Compared with the control group,the cell cycle of the A549 cells did not change significantly,and the expression of Cyclin D1,CDK4 decreased but P16,Rb increased when PSMA7 was upregulated. Compared with the control group,the proportion of phase G0/G1,G2/M of the A549 cells decreased and phase S increased,and the expression of Cyclin D1,CDK4 increased but P16,Rb decreased when PSMA7 was downregulated. There was statistical significance for those results. Conclusion: PSMA7 could affect the expression of Cyclin D1,CDK4,P16,Rb protein level of RB pathway in A549 and promoted the A549 cells into phase S when it's downregulated.