野生型p14ARF基因转染对人肺癌细胞生长抑制的这实验研究

目的 探讨外源性p14ARF基因能否对体内外生长的肺癌细胞起抑制增殖的作用。方法 选择4株具有不同基因背景的肺癌细胞系,转染人的野生型p14ARF基因。通过RT-PCR、免疫组化及Westernblot杂交检测,筛选出同时表达p14ARF mRNA及蛋白的阳性克隆。以母细胞及转染空载体细胞为对照,比较它们的增殖、细胞周期分布、裸鼠体内成瘤性以及形态学上的差异。结果 三株具有野生型p53基因的肺癌细胞在转染p14ARF基因后,细胞周期被显著阻滞于G1或G1/G2期,增殖受到抑制。结论 外源性野生型p14ARF基因转染能抑制人肺癌细胞的增殖,p14ARF基因是人体肺癌基因治疗的理想侯选基因。

Transfection of wild-type p14ARF gene leads to growth inhibition of human lung cancer cell lines

Objective To examine whether wild type p14ARF gene is a candidate suppressor gene for lung cancer. Methods Human lung cancer cell lines having various endogenous backgrounds in INK4a, p53 and Rb genes were used as the recipients of the wild type p14ARF gene. The expression of p14ARF mRNA and protein was detected with RT PCR, immunohistochemistry and Western blot after G418 selection. Clones which expressed both p14ARF mRNA and protein were identified and selected for further experiements. By comparing with the parental and negative control cells treated with empty vectors, the effects of exogenously transfected p14ARF on cell division rate, cell cycle distribution and morphologic alteration were analyzed. In vivo evaluation of the growth rate was also made with the experiment of nude mice tumor formation. Results Upon transfection with p14ARF gene, cells were arrested at G1 or G1/G2 phase of cell cycle in three wtp53 lung cancer cell lines and their proliferation rates were also inhibited. Conclusion Human wild type p14ARF gene has suppressive effect on abnormal proliferation of lung cancer cells, especially in some wtp53 lung cancer cells, and it might be an ideal candidate for gene therapy of human lung cancer.