N端截短的视网膜母细胞瘤基因对人骨肉瘤细胞株SaOS-2生长的影响  

Effect of N - terminal truncated Rb^(66) expression on the growth of human osteosarcoma cell line SaOS -2.

作　　者：李扬[1] 乔慧[1] 邓家德[2] 张萌[1] 朱全胜[1] 丘钜世[1] 

机构地区：[1]中山大学中山医学院病理教研室,广州510089 [2]广州市第一人民医院检验科

出　　处：《中国骨肿瘤骨病》2004年第2期95-99,共5页Chinse Journal Of Bone Tumor And Bone Disease

基　　金：国家自然科学基金(3997028);中山医科大学"211工程"重点学科建设基金(No:98028)资助

摘　　要：目的 研究N端截短的视网膜母细胞瘤基因(Rb^(66))对骨肉瘤细胞Saos-2生长的影响。方法 构建携带Rb^(66)cDNA片段的真核细胞表达载体pcDNA3-1-Rb,用脂质体LIPOFECTAMIME^(TM)2000将真核表达载体pcDNA3-1-Rb、空载体对照pcDNA3-1转染至SaOS-2细胞中。RT-PCR及免疫细胞化学检测转染后SaOS-2中Rb^(66)mRNA及蛋白的表达;观察细胞形态的改变;测定细胞倍增时间及生长曲线的改变;流式细胞仪检测对细胞周期的影响。结果 成功地将N端截短的Rb^(66)cDNA导入骨肉瘤细胞SaOS-2中,并在mRNA及蛋白水平获得了表达。转染后的SaOS-2细胞胞浆展开,细胞核浆比例变小;Rb^(66)导入后,SaOS-2细胞生长减慢,群体倍增时间延长,细胞周期呈现G_1期阻滞。结论 Rb^(66)可通过阻止细胞G_1→S期进展而抑制骨肉瘤细胞SaOS-2的增殖。Objective To investigate the effect of truncated Rb66 expression on the growth of human os-tesarcoma cell line SaOS -2. Method The Rb66 expression vector PCDNA3 - 1Rb was constructed by insertin a 1. 65kD Rb cDNAsequence into the plasmid PCDNA3 - 1. SaOS -2 cells were transfected with PCDNA3 - 1 - Rb or PCDNA3 - 1 by lipofectamin?2000 , the pRb66 mRNA expression was detected by RT - PCR and protein expression by immunocytochemistry. Growth features were observed through the population doubling time and growth curve. Cell cycle analysis was performed by flow cytometry. Result Rb66 was successfully transfected into osteo-sarcoma cell line. SaOS - 2 and expressed in mDNA and protein level Rb66 - transfected cells grew slowly and the population doubling time prolonged. Compared with unransfected SaOS - 2 and SaOS - pcDNA3 - 1 , more SaOS -2 - Rb cells were arrested in G1/G0 phase. Conclusion Rb can suppress the growth of ostesarcoma cell line SaOS -2 through influencing cell cycle G1 - S transition.

关 键 词：N端截短 视网膜 母细胞瘤基因 骨肉瘤细胞株 SaOS-2生长 细胞周期 瘤细胞 

分 类 号：R738.1[医药卫生—肿瘤]