摘要
目的 研究p16基因对人鼻咽癌细胞系CNE 2细胞增殖、凋亡的影响。方法 采用脂质体介导的基因转染方法 ,将野生型p16基因、空载体转入该基因突变的CNE 2细胞中 ,同时设置不加任何质粒的CNE 2细胞作为对照。结果 脂质体转染、G418筛选所获得的抗性克隆 ,经免疫组化检测证实转入p16基因的CNE 2细胞有p16蛋白表达 ,细胞生长速度明显减慢 ;FCM检测显示 ,细胞周期主要阻滞在G1期 ,S期细胞明显减少 ;荧光染色和TUNEL两种检测法均表明 ,导入p16基因的CNE 2细胞的凋亡明显增加 ;免疫组化检测证实 ,p16基因导入后出现p5 3蛋白表达与之相伴随。结论 外源性p16基因导入CNE 2细胞后可有效地表达并导致CNE 2细胞G1阻滞 ,诱导凋亡发生。其机理可能与野生性p5
Objective To study the effects of exogenous p16 gene on proliferation and apoptosis of human nasopharyngeal carcinom cell line CNE 2.Methods Exogenous p16 gene was transfected into CNE 2 cells by lipofectin. After G418 selection, we obtained p16 stably expressed NPC cell clones ( CNE 2 p16), and compared it with the mock tranfected clones(CNE 2 vect). Cell cycle was determined by flow cytometry, and cell apoptosis was examined by fluorescent(Hoechst) staining and TUNEL. P16 protein and p53 protein were stained immunohistochemically with relevant antibodies.Results Exogenous p16 gene was successfully expressed by CNE 2 cells. The growth of CNE 2 p16 cells was inhibited significantly. In CNE 2 p16 cells, cycle arrest in G 1 phase, with decrease in percentage of S phase cells. Apoptotic index of CNE 2 p16 was significantly increased. CNE 2 p16 cellls were positive for p53 protein that it was negative in CNE 2 cells.Conclusion Exogenous p16 inhibits proliferation and induces apoptosis of human nasopharyngeal cancer CNE 2 cell line. It may be related to the up regulation of wild type p53 gene.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2000年第3期192-194,共3页
Chinese Journal of Oncology
关键词
鼻咽肿瘤
P16基因
基因转染
细胞凋亡
Nasopharyngeal neoplasms
p16 gene
Gene transfection
Apoptosis
