人脑胶质瘤低氧诱导因子-1α的基因表达  

Expression of hypoxia-inducible factor-1α in humanglioma

作　　者：于如同[1] 孙兵[1] 赵惠仁[1] 姜曙[2] 高立达[2] 倪鸣山[1] 贺民[2] 毛伯镛[2] 

机构地区：[1]徐州医学院附属医院神经外科,221002 [2]华西医科大学附属第一医院神经外科

出　　处：《中华实验外科杂志》2001年第4期325-326,共2页Chinese Journal of Experimental Surgery

基　　金：江苏省科委资助项目 (DJ9950 1 )

摘　　要：目的　从人脑胶质瘤中克隆低氧诱导因子 1α(HIF 1α)cDNA ,探讨其对人脑胶质瘤发生、发展的作用。方法　从人脑胶质瘤组织中提取总RNA ,以逆转录 多聚酶链反应 (RT PCR)方法扩增出全长 2 481bp的HIF 1αcDNA ,经NheⅠ和BamHⅠ双酶切后 ,将其与表达载体pWAV1连接 ,转化入E .coliJM 10 9,建立HIF 1α的cDNA克隆 ;测序后与大鼠HIF 1αcDNA序列进行同源性分析。结果　RT PCR产物进行琼脂糖凝胶电泳 ,可见一清晰特异扩增带 ,长 2 .5kb ,HIF 1αcDNA经筛选、双酶切鉴定后 ,可见 5 .3kb的载体片段及 2 .5kb的插入片段。进行序列分析 ,与鼠有 90 %的同源性。结论　HIF 1α已成功从人脑胶质瘤中得到克隆和序列分析。Objective To obtain the cDNA of hypox ia-inducible factor-1α(HIF-1α) in human glioma and to investigate its clini cal significance in the development of human glioma. Methods The total RNA was extracted from human glioma and the intact cDNA of HIF-1α was amplified by RT-PCR. The results of RT-PC R were digested by NheⅠand BamHⅠ. The cDNA was cloned into pWAV1 vector and tr ansformed into E. coli JM109 to establish the cDNA clone of HIF-1α. The sequen ci ng was identified by ABI377 automatic sequencing instrument and the homology bet ween the cDNA sequence of rat and human HIF-1α was analyzed. Results After the product of RT-PCR was identified by ASEM, a 2.5 kb amplified ladder was seen. After the recombinant plasmids were ev aluated by NheⅠand BamHⅠagain, the 5.3 kb vector fragments and 2.5 kb insertin g fragments were expressed. The sequencing was identified successfully and the s equence analysis showed that the homology of cDNA between rat and human was 90%. Conclusion HIF-1α cDNA has been cloned in human gliom a and its sequence analyzed.

关 键 词：低氧诱导因子-1Α 神经胶质瘤 CDNA克隆 序列分析 脑瘤 

分 类 号：R739.41[医药卫生—肿瘤] R730.264[医药卫生—临床医学]