肾癌抑制性消减文库构建及差异表达基因鉴定  

Construction and screening of suppression subtractive hybridization library of renal cell carcinoma

作　　者：张勇[1] 艾军魁[1] 张志文[2] 叶雄俊[1] 朱宏建[1] 辛殿旗[1] 梁丽莉[1] 那彦群[1] 郭应禄[1] 

机构地区：[1]北京大学第一医院泌尿外科研究所,100034 [2]北京大学医学部生理学系

出　　处：《中华外科杂志》2003年第2期90-92,共3页Chinese Journal of Surgery

基　　金：国家自然科学基金资助项目 (39870 841);首都医学发展科研基金资助项目(ZD19980 3);北京大学人类疾病基因中心科研基金资助项目 (2 0 0 0 A 18)

摘　　要：目的　应用抑制性消减杂交技术筛选人肾癌差异表达基因。　方法　以 786 0和HK 2为消减杂交对象构建人肾癌抑制性消减文库 ,挑选阳性克隆进行测序及GenbankBLAST分析。结果　文库包含 3 62个有插入片段的阳性克隆 ,随机分析 5 0个克隆 ,其中 2个为新基因 ,另 48个源于3 6个已知基因 ,这些差异表达基因与肿瘤细胞的转录、翻译、增殖、凋亡、代谢、信号转导、血管新生、膜受体表达异常等有关。　结论　该文库质量可靠 ,筛选出包括低峰度和新基因在内的肾癌差异表达基因 ,为进一步研究肾癌发生。Objective To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC). Methods Poly A + RNA was isolated from RCC lines 786 O(tester) and renal cell(RC) lines HK 2(driver), respectively. SSH procedure was performed according to the protocol of the PCR Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT Adv vector and transformed E.coli TOP10F′. All positive clones picked out were digested and some of which were sequenced. Results The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0 3 to 0 9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes. Conclusion The quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC.

关 键 词：肾肿瘤 基因文库 基因表达 抑制性消减杂交 

分 类 号：R737.11[医药卫生—肿瘤]