p53RFP对SW480细胞增殖、细胞周期及凋亡的影响  

Effect of p53RFP on the proliferation,cell cycle and apoptosis of SW480 cells

作　　者：裴静娴[1] 王月刚[2] 程诚[2] 刘城[2] 吴平生[2] Pei Jingxian;Wang Yuegang;Cheng Cheng;Liu Cheng;Wu Pingsheng(Department of Cardiology,Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260;Department of Cardiology,Nanfang Hospital of Southern Medical University,Guangzhou 510515,China)

机构地区：[1]广州医科大学附属第二医院心血管内科,广东广州510260 [2]南方医科大学南方医院心血管内科,广东广州510515

出　　处：《广州医科大学学报》2022年第3期17-20,共4页Academic Journal of Guangzhou Medical University

基　　金：国家自然科学基金青年基金资助项目(30800462)。

摘　　要：目的:观察p53RFP过表达对SW480细胞增殖、细胞周期及凋亡的影响。方法:利用脂质体介导法将p53RFP表达载体转染SW480细胞,Western blot检测p53RFP蛋白表达水平,实时定量RT-PCR检测细胞周期相关蛋白mRNA表达。采用CCK-8检测p53RFP表达上调对SW480细胞增殖的影响、PI染色流式细胞术进行细胞周期分析、Annexin V-FITC/PI双染流式细胞术检测细胞凋亡。结果:p53RFP转染后48 h、72 h、96 h细胞增殖受到显著抑制(P<0.001),p53RFP转染后48 h细胞G2期细胞含量显著高于空载转染组(P<0.05),细胞凋亡率亦显著高于空载对照组(P<0.05)。p53RFP过表达可诱导p21 WAF1/CIP1mRNA表达增加(P<0.05),对CyclinB1、CyclinD1、CDK1 mRNA表达无显著影响。结论:p53RFP过表达可抑制SW480细胞增殖、介导G2期阻滞、诱导细胞凋亡。Objective:To investigate the effect of p53RFP overexpression on the proliferation,cell cycle and apoptosis of SW480 cells.Methods:p53RFP expression vector was introduced into SW480 cells by liposome-mediated transfection.The protein expression level of p53RFP was detected by Western blot,and the mRNA expression of cell cycle-related proteins by real-time quantitative RT-PCR.CCK-8 was used to assess the effect of up-regulated p53RFP expression on proliferation of SW480 cells,Flow cytometry was used to analyze the cell cycle with PI staining,and to detect cell apoptosis with Annexin V-FITC/PI double staining.Results:The cell proliferation was significantly inhibited at 48h,72h,and 96h after transfection of p53RFP expression vector(P<0.001).At 48h after p53RFP transfection,the number of cells at G2 phase was significantly higher than that in the empty vector transfection group(P<0.05),and so was the rate of cell apoptosis(P<0.05).p53RFP overexpression induced an increase in the expression of p21WAF1/CIP1 mRNA(P<0.05),but had no significant effect on the mRNA expression of CyclinB1,CyclinD1,and CDK1.Conclusion:Overexpression of p53RFP can inhibit the proliferation of SW480 cells,mediate cell arrest at G2 phase,and induce cell apoptosis.

关 键 词：p53RFP 增殖 细胞周期 凋亡 

分 类 号：R3[医药卫生—基础医学]