野生型p53基因转移抑制兔心肌细胞生长的实验研究  

Inhibition of wild-type p53 gene on the growth of cultured rabbit cardiomyocytes

作　　者：滕宗艳[1] 张一娜[1] 尹新华[1] 凌虹[2] 

机构地区：[1]哈尔滨医科大学附属第二医院,黑龙江哈尔滨150086 [2]哈尔滨医科大学微生物教研室,黑龙江哈尔滨150086

出　　处：《中国地方病学杂志》2002年第5期348-350,共3页Chinese Jouranl of Endemiology

基　　金：黑龙江省教委基金资助项目 (19990 6)

摘　　要：目的　探讨野生型 p5 3基因转移对兔心肌细胞生长的抑制作用及 p5 3基因用于心肌肥厚基因治疗的可能性。方法　将腺病毒介导的野生型 p5 3基因转入体外培养的乳兔心肌细胞 ,应用生化染色方法、Westernblot杂交检测野生型 p5 3基因的转染效率及表达结果 ,采用透射电镜、MTT增殖抑制实验 (MTT)及流式细胞仪细胞周期分析方法研究野生型 p5 3基因对细胞生长的抑制作用。结果　重组腺病毒能有效地将野生型 p5 3基因转入心肌细胞中 ,并且检测到 p5 3基因表达 ;转入的 p5 3基因导致心肌细胞收缩、体积变小 ;MTT结果显示 ,野生型 p5 3基因能够抑制心肌细胞生长 ,且生长抑制率呈现时间和剂量依赖性 ;心肌细胞增殖周期结果显示 ,细胞G0 - G1 期 DNA百分含量明显高于对照组 ,提示发生 G1 期阻滞。结论　野生型 p5 3基因能高效转染体外培养的心肌细胞 ,对心肌细胞生长有明显的抑制作用 ,其抑制作用的机理是由于野生型 p5 3基因影响心肌细胞的增殖周期 ,以上结果为心肌肥厚的基因治疗提供了重要的实验依据。Objective To study the inhibition of wild type p53 gene on the growth of cultured cardiomyocytes and the possibility of p53 gene therapy for cardiomegaly.Methods We transfered wild type p53 gene mediated by recombinant adenovirus vector into cardiomyocytes.The efficiency of transfection and the expression were detected by X gal staining and Western blot.Inhibition was verified by MTT, transmission electronic microscope and flow cytometry(FCM).Results p53 gene could be quickly and effectively transfered into the cardiomyocytes.The p53 protein could be detected in the transfered cells.Wild type p53 gene could significantly inhibit growth of cardiomyocytes,the inhibition depended on dosage and time.Result of FCM showed that wild type p53 gene could affect cell cycle and provide G 1 arrest.Conclusions Wild type p53 gene mediated by recombinant adenovirus vector can effectively transfect cultured cardiomyocytes,which can inhibit cultured cardiomyocytes growth.This study provides an important reliance for gene therapy of cardiomegaly.

关 键 词：实验研究 野生型P53基因 心肌细胞 腺病毒 心肌肥厚 基因治疗 

分 类 号：R542.2[医药卫生—心血管疾病]