转染p21基因对人动脉平滑肌细胞生长的作用  

Effect of p21 gene transfection on human aortic smooth muscle cell lines

作　　者：姜兰兰[1] 方五旺[1] 林丙来[1] 

机构地区：[1]安徽省芜湖市第二人民医院心内二科,安徽芜湖241000

出　　处：《安徽医药》2010年第3期311-313,共3页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨转染p21基因对人动脉平滑肌细胞(HASMC)的影响,以探讨转染p21基因实现抗冠状动脉支架内再狭窄的作用,为冠状动脉支架内再狭窄的治疗提供新思路。方法以Lipofectamine 2000脂质体介导p21基因转染HASMC株;免疫组化法检测转染p21基因后,其编码蛋白在HASMCs的表达情况;描绘p21基因转染后HASMCs的增殖曲线;WST-1法测定OD值,计算细胞生长抑制率及流式细胞仪检测p21基因转染对HASMCs凋亡的影响。结果免疫组化法显示p21基因成功转入HASMCs后,其编码的蛋白能在细胞质内进行高表达;转染p21基因的HASMCs在体外生长曲线较对照组明显降低;WST-1法显示转染p21基因的HASMCs细胞活力与对照组相比有显著性差异(P<0.01);流式细胞仪检测发现转染p21基因的HASMCs细胞凋亡率达40%以上,且显著高于对照组。结论成功将p21基因转入HASMCs,其编码蛋白可能参与了抑制细胞生长和诱导细胞凋亡作用,提示p21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 inhibition and apoptosis effect of p21 gene transfection on human aortic smooth muscle cell（HASMC） lines,in order to discuss p21 gene transfection into HASMCs seeking a new way for prevention of restenosis after coronary artery stent insertion（CASI）. Methods p21 express vector or pcDNA3.1 （＋） blank vector was transfected,into HASMC lines and normal HASMCs were used as normal control. How was the protein of p21 gene expressed by immunohistochemistry assay？ The inducing proliferation ,inhibition and apoptosis effects of p21 gene on cell growth was described by cell growth curve, WST - 1 assay and flow cytometry（FCM） analysis. Results Immunohistochemistry assay showed that the protein of p21 gene was expressed in the cytoplasm of HASMCs. Expression of p21 gene significantly inhibited HASMC proliferation in vitro, and the growth of HASMC/p21 was notably lower than that of control. WST- 1 test showed that the difference of cell viability between HASMC/p21 and control cells was also significant（ P 〈 0.01 ）. FCM showed that the rate of HASMC/p21 apoptosis was notably higher than that of control cells,and above 40%. Conclusions The findings showed that the transfection of p21 gene obviously induced cellular inhibition and apoptosis in HASMCs, which suggested that p21 gene transfeetion into HASMCs technique can be effective means in the prevention and therapy of restenosis after CASI.

关 键 词：P21基因 转染 HASMC 凋亡 

分 类 号：R543[医药卫生—心血管疾病]