摘要
背景与目的:p53基因突变是肺腺癌发生中的常见的分子生物学事件,第245位点突变为其突变热点之一,因此研究p53基因第245位点突变可能为肺腺癌的诊断、治疗提供一定的理论基础。本研究旨在研究肺癌常见的245位点突变对p53缺失的人肺腺癌细胞系H1299的影响。方法:应用流式细胞术观察脂质体瞬时转染p53基因第245突变体(G245V)对细胞凋亡的影响,同时用稳定转染观察克隆形成率和生长曲线的不同。结果:转染G245V突变体的H1299细胞凋亡平均数低于转染野生型p53基因的细胞,差异有统计学意义(P<0.001);大中克隆形成数量比导入野生型p53基因的细胞大中克隆形成数量明显增多,差异有统计学意义(P<0.05);细胞的生长速度也明显加快。结论:p53基因第245位点发生突变后基本丧失了野生型p53的功能,是肿瘤细胞增殖的原因之一。
Background and purpose:The mutation of the p53 gene could be a common molecular biological event during the process of lung adenocarcinoma because 245 site was a hot site for p53 gene mutation.A study of 245 site's mutation of p53 may provide the basic theories for the diagnosis and treatment for lung adenocarcinoma.This study aimed to investigate the influence of p53 245 site's mutation on p53-deficient human lung cancer H1299 cell line in vitro.Methods:H1299 cells were transiently transfected with wide type p53 and G245V p53 plasmid.Flow cytometry was performed for cell apoptosis analysis.H1299 cells were also stably transfected with wide type p53 and G245V p53 plasmid.The G418-selected cells were assayed for colony-forming efficiency,and the transfected cells were examined using MTT for the purpose of drawing the growth curve and evaluating cell growth.Results:In the experimental study of cell apoptosis,we could see that the apoptosis rate of H1299 cells transfected with G245V-p53 gene appeared significantly less than in cells transfected with wide type p53 gene(P<0.001),similar happened to those transfected with pcmv plasmid.The colony-forming efficiency showed that there were more cells transfected with G245V-p53 gene than those transfected with wide type p53 gene(P<0.05).The growth curve indicates that cells transfected with G245V-p53 gene grew faster than those transfected with wide type p53 gene.Conclusion:Wild type p53 gene loses all function when mutation happens to the site 245.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2011年第3期177-181,共5页
China Oncology
