ΔNp73基因对不同p53基因状态肺癌细胞系体外生长的影响

Effect of exogenous ΔNp73 gene on the growth of human lung adenocarcionoma cell lines with different p53 status

目的 探讨 p73基因N末端转录活化区缺失的异构体ΔNp73转染对不同p5 3基因状态肺癌细胞系体外生长的影响。方法 利用脂质体将ΔNp73α基因分别转染P5 3蛋白表达阴性与表达野生型P5 3蛋白的肺癌细胞系H12 99和A5 4 9,筛选出稳定的表达株 ,四甲基偶氮唑盐 (MTT)法检测细胞生长情况 ,平板克隆形成率观察细胞增殖能力的变化 ,流式细胞术分析细胞周期及细胞凋亡率。结果 ΔNp73α基因转染使H12 99和A5 4 9细胞生长明显增快 ,克隆形成率明显提高 ,细胞凋亡率明显降低但对细胞周期没有明显的影响。结论 ΔNp73基因在肺癌的发生发展过程中发挥癌基因的功能 ,并且其作用不受 p5 3基因状态的影响。

Objective To study the effect of ΔNp73 gene on human lung adenocarcionoma cell lines H1299 and A549 with different p53 status.Methods The ΔNp73α expression vector (pcDNA3-ΔNp73α) was respectively transfected into H1299 (loss of P53 protein expression) and A549 (positive wtP53 protein expression). The postive cell clone was screened by G418. The cell growth curve was analyzed by MTT method. The cell proliferation and survival capacity were observed by colony formation assay. Cell apoptosis percentage and cell cycle phase distribution were measured by flow cytometric assay (FCM).Results The growth and colony forming efficiency of H1299 and A549 cells were significantly enhanced after transfected with ΔNp73α. Furthermore, ΔNp73α upregulation inhibited apoptosis but had no effect on cell cycle.Conclusion Overexpression of ΔNp73α protein results in malignant transformation of human lung adenocarcionoma cell lines with different p53 status. It supports an oncogenic function of ΔNp73α.