转染p16基因对人肺腺癌细胞放射效应的影响

Effect on radiosensitivity of human lung adenocarcinoma cells transfected with wild-type p16 gene

目的 :探讨p16基因与人肺腺癌细胞放射效应的关系。方法 :免疫组化、Westernblot方法检测人肺腺癌细胞株Anip973、AGZY83a的p16蛋白表达 ,FuGene转染方法把 p16表达载体转入这两个细胞株 ,进行细胞存活曲线、流式细胞仪 (FCM )分析细胞周期分布。结果 :p16蛋白表达AGZY83a为 87.4 % ,明显高于Anip973(5 2 % ) ,细胞存活曲线显示低表达的Anip973与转染后Anip973p16的Dq值有显著性差异 (P <0 .0 5 ) ,D0 值间无显著差异 (P >0 .0 5 )。p16蛋白高表达的AGZY83a与AGZY83ap16间D0 值、Dq值均无显著性差异 (P >0 .0 5 ) ;流式细胞仪测定细胞周期结果显示 ,Anip973转染 p16基因后出现G2 M期比例增加 ,S期比例减少。结论 :转染 p16基因后可能通过改变细胞周期分布及抑制细胞对照射后的亚致死性损伤修复 ,来改变p16蛋白低表达的人肺腺癌细胞株Anip973的放射效应 ,而对于高表达的AGZY83a则无显著影响。

Purpose:To investigate the correlation between p16 gene and the radiation effect on human lung adenocarcinoma cell lines. Methods:We examined the expression of p16 protein of human lung adenocarcinoma cell lines with immunohistochemistry and Western blot analysed, transferred the p16 expression vector into the two cell lines with Fu Gene Kit ,and analysed the cell survival curve. Flow cytometry(FCM)was used to show the distribution in cell cycle.Results:The expression of p16 in AGZY83a was 87.4%,which was markedly higher than Anip973(5.2%);The Dq of the two cell lines was significantly different ( P <0.05), while the D 0 of Anip973 and Anip973 p16 was similar ( P >0.05). there was no significant difference between AGZY83a and AGZY83a p16 in the D 0 and Dq value ( P >0.05); The result of FCM indicates that the cells transfected with p16 changed in cell cycle distribution ; proportion of G 2 M phase was increased, while that of S phase was decreased.Conclusions:Introduction of p16 gene into low expression human lung adenocarcinoma cell line Anip973 alters the radiation effect in a way which perhaps was correlated with the inhibition of sublethal damage repair and the cell cycle distribution change, but not in the high expression cell line AGZY83a.