mRNA差异显示法克隆前列腺癌相关基因  

Cloning of the prostate cancer related genes with mRNA differential display

作　　者：杨学贞[1] 周利群[1] 周健[2] 王夏[2] 杨帆[3] 张志文[4] 那彦群[1] 顾方六[1] 郭应禄[1] 

机构地区：[1]北京大学第一医院泌尿外科北京大学泌尿外科研究所,100034 [2]中国科学院微生物研究所 [3]北京大学人民医院检验科 [4]北京大学医学部生理学系

出　　处：《中华泌尿外科杂志》2003年第8期545-547,共3页Chinese Journal of Urology

基　　金：国家自然科学基金资助项目 ( 3 980 0 173 ) ;国家教委留学归国人员科研启动基金资助项目 ( 98 -2 0 0 0 )

摘　　要：目的　克隆前列腺癌相关基因。　方法　应用改进的mRNA差异显示技术 ,从前列腺癌患者和正常成人前列腺组织中获得两者的差异片段 ,即表达序列标签 ,克隆测序分析 ,采用逆转录聚合酶链式反应 (RT PCR)方法证实。　结果　正常成人和前列腺癌患者的前列腺组织存在明显的基因表达差异。在GenBank分析的 7个差异显著的片段中 ,5个为新基因片段 ,1个与基因聚集素(clusterin) 10 0 %同源 ,在前列腺癌组织中高表达 ;另一个与超氧化物歧化酶 1(SOD1) 97%同源 ,在正常前列腺组织中高表达。RT PCR结果证明前列腺癌组织及前列腺癌细胞株中 ,clusterin与内参基因β actin的相对表达量明显高于正常前列腺组织。 　结论　 5个新基因片段、clusterin和SOD1基因表达在前列腺癌的发生、发展中可能发挥重要作用。Objective Cloning of some specific genes related to the prostate cancer. Methods Specimen from a patient with prostate cancer and from a normal adult were studied by the improved mRNA differential display and the differentially expressed sequence-tags were cloned,sequenced and analized.Reverse transcriptive-polymerase chain reaction(RT-PCR) was used to examine the expression level of Clusterin in prostate cancer tissues,prostate cancer cell line and normal prostate tissues. Results Significant difference was observed between the two kinds of tissues in gene expression and seven differentially expressed sequence-tags were found through analysis in GenBank.Among them,five were proved to be new gene tags,one shared 100% homology with Clusterin,which was highly expressed in the prostate cancer tissue and the other one shared 97% homology with superoxide dismutase 1 (SOD1),which was highly expressed in the normal prostate tissue.It was proved by RT-PCR that to compare with the internal marker gene β-actin,the expression level of clusterin in prostate cancer is much higher than that of normal prostate. Conclusions Five new gene tags,clusterin and SOD1 were found to be differentially expressed between tissues of prostate cancer and normal prostate and may play important roles in carcinogenesis and development of prostate cancer.

关 键 词：前列腺癌 MRNA差异显示技术 基因克隆 逆转录聚合酶链式反应 基因聚集素 超氧化物歧化酶 

分 类 号：R737.25[医药卫生—肿瘤]