p16基因转染对胰腺癌细胞生长及辐射敏感性的研究

目的 探索p16基因转染对胰腺癌细胞生长辐射敏感性的作用。方法 将外源野生型p16基因连接到pCDNA3.1^＋载体构建pCDNA3.16＋ -p16重组质粒,利用Lipofectamine^TM介导转染胰腺癌JF305细胞,筛选阳性克隆。照射后,RT-PCR检测p16基因表达,Western blot检测蛋白表达;用MTT法测定细胞生长曲线和肿瘤细胞杀伤率。结果 转染p16基因的JF305细胞有外源p16基因的整合及表达,生长速度明显减少,对辐射的敏感性增强。结论 导入外源野生型p16基因可抑制胰腺癌细胞恶性增殖,提高胰腺癌细胞对辐射的敏感性。

Effects of p16 gene on the growth and radiation sensitivity of human pancreatic carcinoma cell line

ObjectiveTo observe the effects of p16 gene transfection on the growth and radiation sensitivity of human pancreatic carcinoma cell lines. Methods Wild-type p16 cDNA was ligated to pcDNA3.1~+ to construct pcDNA3.1-p16 plasmid. Wild-type p16 gene was transfected into JF305 pancreatic carcinoma cell lines by lipofectamine TM, then the clones were randomly selected. Being irradiated,expression of exogenous p16 gene were detected with RT-PCR and Western blot. MTT assay was used to determine the curve of cell growth and the rate of damaged cells by irradiation. Results The integration and expression of p16 gene was demonstrated in JF305 cell after p16 transfected.The cell growth velocity was found obviously decreased and radiation sensitivity increased. ConclusionThe transftection of p16 gene can inhibit malignant generation and increase sensitivity of human pancreatic carcinoma cells to radiation.