摘要
进行ＭＸＲ７基因ｃＤＮＡ的克隆鉴定并检测其在人正常组织和肿瘤组织中的表达。方法以人肝癌组织的ｃＤＮＡ为模板，应用ＰＣＲ方法合成ＭＸＲ７基因ｃＤＮＡ，连接于融合蛋白表达载体ｐＧＥＸ－５Ｘ－１，构建成符合读框的融合基因；应用Ｎｏｒｔｈｅｒｎｂｌｏｔ对正常人１２种不同组织和术前做治疗的７例非肝脏肿瘤组织及３０例肝癌和癌旁肝组织、１２例正常肝组织中ＭＸＲ７基因的表达进行检测。结果ＭＸＲ７ｍＲＮＡ在人肝脏等１２种正常组织及７例其它部位肿瘤组织中均不表达；在人肝癌中呈高表达，而癌旁肝组织中呈低表达，其阳性表达率分别为７６．７％和１３．３％。结论ＭＸＲ７．基因在人肝癌组织中的高水平表达具有普遍性和特异性，提示ＭＸＲ７基因可以作为肝癌的生物学标志物。
Objective To clone and identify the whole cDNA of MXR7 gene and to fied out its exprmsion in human normal and tumor tissues. Method The DNA primers were designed and syntheised acordingto the whole cDNA sequence of MXR7 gene. The cDNA of human HCC was taken as template while the cDNA of MXR7 gene was synthesized by polymerase chain reaction (PCR), Recombinant DNA conforming toreading frame was constructed by connecting purified PCR product of the cDNA of MXR7 gerre with expression vector pGEX-5X-1 of fusion protein. The plasmid MXR7/pGEX-5X-1 was identified by sequencing. Us ing 32 P labeled MXR7cDNA as probe, MXR7mRNA expression was detected by Northern blot analysis in 12different human normal tissues, 7 preoperatively untreated non-liver tumor tissue, 30 preoperatively UntreatedHCC and the corresponding surrounding non-cancerous hepatic tissues samples, 12 normal liver tissues samplesconfirmed pathologically. Results Enzyme digest and sequence analysis confirmed that the insertion sequencein vector PGEX-5X-1 was the same as the cDNA sequence of MXR7 gne. Northern blot analysis showed nodetectable expression of MXR7 mRNA in 12 human normal tissues including liver, 7 non-HCC tumor tissuesand 12 normal liver tissues and the frequencies of MXR7 mRNA expression in HCC and the coresponding sur rounding non-cancerous hepatic tissues are 76.6% and 13.3%, respectivdy. Concluslon MXR7 mRNAhigh expression in human HCC was common and specific, suggesting MXR7 mRNA may serve as a tumorbiomarker for HCC.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
1999年第2期144-146,F004,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金
美国Sugen高技术公司基金
关键词
肝肿瘤
MXR7基因
基因克隆
基因表达
Carcinoma, hepatocdlular Gene, MXR7 Gene cloning Gene expression，Normal tissue Tumor tissue
