重组腺病毒介导的人AWP1基因转移对人源肝癌细胞增殖的影响

Influence of human AWP1 recombinant adenovirus on proliferation of hepatocarcinoma cells

目的以重组腺病毒作为AWP1(associated with PRK1)的转基因载体,研究AWP1对人肝癌细胞增殖的影响,为进一步研究AWP1基因功能提供依据。方法重组腺病毒质粒pAd-flag-AWP1转染293细胞进行重组腺病毒的包装和扩增,并进行PCR鉴定。用空病毒Ad-Null和重组腺病毒Ad-flag-AWP1分别感染人源肝癌细胞SMMC-7721,并于感染后24 h、72 h、第5天和第7天收集细胞计数,绘制细胞生长曲线,计算细胞倍增时间。结果重组腺病毒Ad-flag-AWP1能够感染SMMC-7721细胞;依据细胞生长曲线计算的SMMC-7721细胞倍增时间分别为:SMMC-7721细胞约为5 d;而Ad-Null感染后其生长被抑制,倍增时间约为6 d;Ad-flag-AWP1感染后,其倍增时间约为3 d。结论重组flag-AWP1腺病毒转染SMMC-7721细胞过表达的AWP1蛋白可能具有促进人肝癌细胞生长的作用。

Objective To explore the influence of human AWP1（associated with protein kinase C related kinase 1） recombinant adenovirus on the proliferation of hepatocarcinoma cells so as to further investigate the biologic function of AWP1.Methods Recombinant adenoviruses Ad-flag-AWP1 were packaged and amplified in 293 cells transfected by recombinant plasmids pAd-flag-AWP1,and were identified by PCR assays.Human hepatocarcinoma cells SMMC-7721 were infected by Ad-Null or Ad-flag-AWP1 and the cell number was counted at 24 h;72 h;5 d and 7 d after the infection.The cell growth curves were painted and the proliferation time was counted.Results According to the growth curves of SMMC-7721,the cell doubling time of SMMC-7721 was five days and that of Ad-Null was six days,but for the cells that were infected by Ad-flag-AWP1,the doubling time was only three days.Conclusion The expression of human AWP1 gene in human hepatocarcinoma cells infected by AWP1 recombinant adenoviruses may be able to accelerate the cellular proliferation.