食管癌相关基因片段的筛选与鉴定

Screening and identification of esophageal cancer associated genes

目的 :从已构建的中国人食管癌特异的消减cDNA文库中 ,初步筛选与鉴定食管癌相关基因片段。方法 :随机挑选 48个片段作反Northern印迹杂交分析 ,将杂交信号有差别的 15个片段送测序公司进行测序 ,用Gen BankBlast程序检索 ,以同源性很小的cDNA片段作为新基因的候选基因 ,观察其在食管癌组织和正常食管粘膜组织中的表达情况。结果 :所获得的 7个序列中 :3y5 9与已知基因同源性很小 ,且 3y5 9在 30例食管癌及其配对正常食管粘膜组织中 ,19例 (6 3.3 % )表达有差别 ,11例 (36 .7% )表达无差别 (P <0 .0 5 )。 3y88与已知基因geminin基因部分同源。另外 5个片段 3y2 0 ,3y14,3y90 ,3y91和 3y92分别与核糖体蛋白RpL7a,TLH2 9蛋白驱动子 ,Hsp70 /Hsp90 organizingprotein ,Keratin6 (KRT6C)和内膜蛋白等已知基因序列高度同源。结论 :3y5 9可能是与食管癌的发生、发展有关的新的候选基因。首次发现 ,geminin基因 ,核糖体蛋白RpL7a,TLH2 9蛋白驱动子 ,Hsp70 /Hsp90 organizingpro tein ,Keratin6 (KRT6C) 。

Aim: Initial screening and identificating the differentially expressed genes associated with esophageal cancer from subtractive cDNA library specific for Chinese. Method: From 100 positive recombinants, 48 clones were randomly picked out for reverse Northern blot analysis.Fifteen clones with markedly differential expression patterns on the blots were sent for sequencing. Homologies of the sequences were compared with the data provided by the GenBank. RT PCR was employed to confirm differential expression of the novel gene candidates in matched esophageal cancer tissues. Results: Among 7 different sequences obtained after analysis of homologies of 15 sequences ,3y59 fragment had little homology to the known sequences in the GenBank,which dramatically overexpressed in 63.3% (19/30) esophageal cancer tissues. 3y88 fragment shared partial homology to the known geminin gene, 5 other cDNA fragments shared great homology to the known genes in GenBank: 3y20 to ribosomal protein RpL7a, 3y14 to TLH29 protein precursor,3y90 to human Hsp 70/Hsp 90 organizing protein,3y91 to Keratin 6,and 3y92 to inner membrance protein. Conclusion : 3y59 might be a novel gene candidate for differentially expressed genes associated with esophageal cancer.Geminin gene and 5 other fragments isolated from esophageal cancer tissues are demonstrated to be associated with esophageal cancer for the first time.