人前列腺癌高低转移细胞株差异表达基因的筛选  

Screening of differently expressed genes in human prostate cancer cell lines with different metastatic potentials

作　　者：宋安萍[1] 廖国宁[1] 吴明富[1] 卢运萍[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤生物医学中心,湖北武汉430030

出　　处：《中华肿瘤防治杂志》2007年第13期961-964,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家重大基础研究项目(973;2002CB513107);国家自然科学基金(30571950)

摘　　要：目的:筛选高低转移能力人前列腺癌细胞株PC3M-1E8和PC3M-2B4间差异表达基因。方法:应用抑制性消减杂交(SSH)技术,对高转移能力人前列腺癌细胞株PC3M-1E8及其同源低转移细胞株PC3M-2B4进行2次消减杂交,第1次SSH实验,以PC3M-2B4细胞株为实验方,PC3M-1E8株为驱动方,构建正向消减文库。第2次实验则以PC3M-1E8株为实验方,PC3M-2B4为驱动方,构建反向消减文库。筛选出来的阳性克隆进行测序,并在GeneBank数据库中进行同源性比较后从中选取若干序列进行实时定量PCR验证。结果:2个消减文库共得到238个阳性克隆,从中各随机选取8个克隆测序并进行同源性分析,发现其中12条序列来自于11个已知基因,另外4条序列为新的未知功能的基因序列标签(EST)。结论:成功构建了高低转移力人前列腺癌细胞株的双向消减杂交文库。OBJECTIVE： To screen genes differentially expressed in two human prostate cancer cells PC3M-1E8 and PC3M-2B4 with different metastatic potentials. METHODS： Suppression subtractive hybridization （SSH） was done twice on human prostate cancer cell line with high potential of metastatic PC3M-1E8 and its synogenetic cell line PC3M-2B4 with low metastatic potential. The first subtraction used PC3M-2B4 as tester and PC3M-1E8 as driver and constructed the forward subtractive library, while the second one interchanged the tester and driver and constructed the reverse subtractive library. The screened clones of both libraries were sequenced and Gene Bank homology search was performed. Some clones were confirmed by quantitative real-time PCR. RESULTS： Two subtractive libraries containing 238 positive clones were constructed. Analysis of 16 sequenced clones randomly picked from two libraries showed that 4 differentially expressed gene fragments were identified as new EST with unknown functions. CONCLUSION： The two subtractive libraries of human prostate cancer cell lines with different metastatic potentials are constructed successfully.

关 键 词：前列腺肿瘤 杂交 遗传 肿瘤转移 克隆 

分 类 号：R737.25[医药卫生—肿瘤]