人端粒酶反转录酶基因诱导人永生化成骨细胞的生物学特性  

Biological characteristics of human immortalized osteoblast cell line induced by human telomerase reverse transcriptase gene

作　　者：殷晓雪[1] 陈仲强[1] 党耕町[1] 郭昭庆[1] 马庆军[1] 

机构地区：[1]北京大学第三医院骨科,北京市100083

出　　处：《中国临床康复》2003年第26期3550-3551,共2页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金(30271305)~~

摘　　要：目的:研究反转录病毒(hTERT)基因诱导的人永生化成骨细胞系Clone5的生物学特性,包括增殖能力、细胞周期和是否恶性变。方法:收集30～31代Clone5和6～7代人成骨细胞(hOB),绘制两种细胞的生长曲线并用流式细胞仪检测细胞周期特征,软琼脂集落实验和裸鼠致瘤实验检测Clone5是否恶性变。结果:由生长曲线计算出细胞倍增时间:hOB:190h;Clone5:62h;曲线显示Clone5的增殖能力明显增强;流式细胞仪检测发现,Clone5细胞周期中G1期细胞占53.46%,S期细胞占28.24%,凋亡率为0.76%,hOBG1期细胞占79.81%,S期细胞占19.76%,凋亡率为33.33%。和hOB相比,Clone5G1期细胞比例减少,S期比例增加,细胞的凋亡率显著降低;软琼脂集落实验未见细胞团形成,裸鼠接种不致瘤。结论:转导hTERT基因的Clone5增殖能力明显增强,凋亡率降低,且未发生恶性变,可用做组织工程的种子细胞和研究正常骨生理、生化的工具细胞。AIM:To investigate the biological characteristics of immortalized human osteob last cell line Clone5 including proliferation,cell cycle and tumorigenesis. METHODS:Thirtieth-31th passage Clone5 and 6th-7th passage hOB cells were exa mined by analysising cell growth curves for proliferate ability, by flow cytomet ry for cell cycles and by soft agar colony formation and tumor formation by inoc ulating cells into nude mice for tumorigenesis. RESULTS:Population doubling time of Clone5 and hOB were approximately 62 hours and 190 hours respectively.G1 and S cells comprised 53.46%and 28.24%respectiv ely,and apoptosis rate was 0.76%in the cell cycle of Clone5 by flow cytometer. G1 and S cells comprised 79.81%and 19.76%respectively, and apoptosis rate was 33.33%in hOB. In the cell cycle of Clone5,compared to hOB, G1 cells were reduce d and S cells increased, the rate of cell apoptosis declined greatly. Furthermor e, Clone5 failed to form colonies in soft agar and did not develop tumors by ino culating cells into nude mice.

关 键 词：人永生化成骨细胞系 Clone5 人端粒酶反转录酶基因 生物学特性 细胞增殖 细胞周期 

分 类 号：R318.17[医药卫生—生物医学工程]