稳定表达外源性p16基因肺癌A549细胞株的建立及鉴定

为构建稳定表达外源性抑癌基因p16的肺癌A549细胞株，用脂质体介导的基因转染方法，借助真核质粒表达载体（pcDNA3)。将抑癌基因p16转移入此基因缺失的人肺癌细胞株A549细胞中，经G418筛选，获得稳定表达的细胞克隆，用逆转录聚合酶链反应（RT-PCR）及免疫组织化学鉴定p16基因的表达，同时对克隆细胞分泌蛋白进行活性检测。结果显示转染p16基因的A549细胞中可以检测到p16mRNA及蛋白的表达，说明建立的p16真核表达载体能在肺肿瘤细胞中分泌表达蛋白，表达P16抑癌蛋白的A549细胞株的建立有助于研究抑癌基因p16在肺癌发生中的作用。

ESTABLISHMENT AND IDENTIFICATION OF A549 CELL LINE WITH STABLEEXPRESSION OFEXOGENOUS P16 GENE

To establish A549 cell line with stably expressing tumor suppressor gene p16,the exogenous tumor suppressor gene p16 was transfected into A549 cells with lipofectin by eukaryotic expressive vector(pcDNA3). We got clone cells after selected by G418. All the clone cells were detected by RT PCR and immunohistochemical staining on mRNA and protein levels. Meanwhile, P16 protein activity were measured by MTT. The Results showed that p16 mRNA and protein were expressed in those transfected A549 cells. It demonstrated that the p16 eukaryotic expression vector can express P16 protein in human lung cancer cells. Exogenous p16 gene may stably expressed in human lung canner cell line A549. The establishment of the cell line can help to study the functions of tumor suppression gene p16 in human lung cancer.;