转染p53基因对培养的人动脉平滑肌细胞的影响  

Effect of wild-type p53 gene modification on human aortic smooth muscle cell lines

作　　者：方五旺[1] 姜兰兰[1] 林丙来[1] 

机构地区：[1]皖南医学院附属芜湖市第二人民医院心内二科,安徽芜湖241000

出　　处：《心脏杂志》2010年第6期856-859,共4页Chinese Heart Journal

基　　金：芜湖市科技局课题资助(2007-68)

摘　　要：目的:探讨转染野生型p53(wtp53)基因对培养的人动脉平滑肌细胞(HASMCs)的影响,为冠状动脉支架内再狭窄的治疗提供新的思路。方法:以Lipofectamine 2000脂质体介导wtp53基因转染HASMCs株后,用免疫组化染色法检测转染wtp53基因后,其编码蛋白在HASMCs中的表达。绘制wtp53基因转染后HASMCs的增殖曲线。用WST-1法测定A值,并计算细胞生长的抑制率。用流式细胞仪检测wtp53基因转染对HASMCs凋亡的影响。结果:免疫组化染色法显示,wtp53基因转染后,在HASMCs的细胞核内高表达;wtp53基因的表达可抑制HASMCs的体外生长。HASMC/wtp53细胞的生长曲线较对照组明显降低;WST-1法显示,HASMC/wtp53的细胞活力与对照组相比有显著性差异(P<0.05)。流式细胞仪检测显示,转染wtp53基因的HASMCs的凋亡率达40%以上,显著高于对照组。结论:转染wtp53基因对HASMCs具有明显的抑制和凋亡作用,提示wtp53基因转染HASMCs技术有可能为人冠状动脉支架内再狭窄的治疗提供一种新的防治策略与手段。AIM： To investigate the inhibitory and apoptosis effect of wild-type p53 （wt p53） gene trans- fection on human aortic smooth muscle cell （HASMC） lines and to explore a new way for prevention of restenosis following coronary artery stenting via wt p53 gene transfection into HASMCs. METHODS： HASMC lines were transfected by wt p53 gene coated by liposome of lipofectamine 2000. The expression of wt p53 protein in HASMC was determined by immunohistochemistry. Cell proliferation and the apoptosis of wt p53 gene on cell growth were detected by cell growth curve, WST-1 assay and flow cytometry. RESULTS： Immunohistochemistry staining showed that the protein of wt p53 gene was expressed in the cellular nucleus of HASMCs. Overexpression of wt p53 gene significantly decreased HASMC proliferation in vitro compared with that of control vector. Growth rate of HASMC/wt p53 （ ＋ ） was significantly lower than control. WST-1 test showed significant differences in cell viability between HASMC/wt p53 and control cells （P 〈0. 05）. FCM showed that the rate of HASMC/wt p53 apoptosis was 〉40%, significantly higher than eontrol cells. CONCLUSION： Transfection of wt p53 gene obviously induces cellular inhibition and apoptosis in HASMCs, suggesting that wt p53 gene transfection into HASMCs can be an effective means for prevention and therapy of restenosis after coronary stenting.

关 键 词：P53基因 基因转染 人动脉平滑肌细胞 细胞凋亡 

分 类 号：R459.9[医药卫生—治疗学]