摘要
用反转录ＰＣＲ从正常人胚胎肺细胞中获得了ｐ２１基因ｃＤＮＡ，将其插入真核表达载体ｐＭＳＣＶｎｅｏ，构建成重组质粒，ｐＭＳ２１，并将其转染至肺癌细胞株Ａ５４９．通过集落形成观察到ｐ２１对肺癌细胞具有明显的抑制作用，经ＲＮＡ狭缝杂交、Ｗｅｓｔ－ｅｒｎｂｌｏｔ分析和免疫细胞化学实验证实这是ｐ２１表达的结果．荷瘤裸鼠实验也进一步证实了ｐ２１对肺癌细胞具有明显的抑制作用．为ｐ２１的深入研究提供了基础．
p21 cDNA was obtained from normal human embryonic lung cells by RT PCR.The recombinant plasmid pMS21 was constructed by inserting the p21 cDNAs into eukaryotic expression vector pMSCV which contains a neomycin resistence marker gene and subsequently transfected into lung cancer cell line A549 with lipofectamine.It was observed that p21 could inhibit clonal formation of lung cancer cell.Slot hybridization of RNA,Western blot and immunocytochemical assay demonstrated that it was the expression of exogenic p21 that led to the above result.Tumor bearing nude mice experiment further showed the inhibitory effect of p21 on lung cancer cell.The data will be of benefit for studying the functional mechanism of p21 and gene therapy for tumor suppression.
