人p16基因是最近发现的一种肿瘤抑制基因.p16蛋白作为一种细胞增殖的负调控因子,在恶性肿瘤发生发展中起重要作用.为了进一步探讨p16基因的抗肿瘤特性,我们通过运用分子克隆技术构建重组人p16基因表达载体(pcDNA3-p16),并通过电穿孔方法将p16基因导入到人肺腺癌NCI-H460细胞中,经G418筛选获得可稳定表达p16的人肺腺癌细胞,观察p16基因对人肺腺癌细胞周期的影响和细胞增殖的作用.结果表明,导入p16基因能使人肺腺癌细胞生长速度较对照细胞明显减慢,克隆形成率下降,细胞周期G1阻滞.结果证明人p16基因能抑制人肺腺癌细胞的生长.本研究为人肺腺癌p16实验性基因治疗提供了实验依据,说明基于恢复p16肿瘤抑制基因功能的基因治疗在治疗人肺腺癌方面有广阔的应用前景.

p16 gene was a tumor supressor gene found recently. The p16 protein is a negative regulator of cell proliferation . Loss of normal p16 function is associated with the development of neoplasms. To detect antitumorigenic effect of p16 on human lung adenocarcinoma cells, we cloned a p16 cDNA into the pcDNA3 vector at the sites of BamHl and Xho I to gain a p16 gene recombinant expression vector plasmid. We then transferred the p16 gene recombinant plasmid into human lung adenocarcinoma cell line (NCI-H460) by electroporation method. After G418 selection we assessed cell growth properties and cell cycle pattern by flow cytometry to G418-resistant clones of NCI-H460-pl6 and NCI-H460-vect respectively. The results show that human p16 gene could suppress the phenotype of NCI-H460 cell line and p16 gene therapy plays a positive role in human lung adenocarcinoma treatment.

R73-36

自然科学基金(39600181)资助