肾癌差异表达基因GYLZ-RCC18的全长克隆及意义

Cloning and identifying of renal cell carcinoma differentially expressed genes

目的 克隆并鉴定肾癌与正常肾组织之间差异表达的基因 ,为研究肾癌发生发展机制提供新的突破口。 方法 应用抑制性消减杂交技术 ,构建人肾癌组织与正常肾组织差异表达的cDNA消减文库 ,并从中克隆鉴定出肾癌特异表达的基因。结果 :构建成功高消减效率的人肾癌组织cDNA消减文库 ,对其中 10个克隆的插入cDNA片段进行测序后经基因库检索表明 10个片段均为未知新序列 ,其中GYLZ RCC18基因为 5个拷贝 ,这提示以上 10个cDNA片段可能来自 6个新基因。差异分析显示GYLZ RCC18在肾癌组织中有明显表达 ,而在正常肾组织中无表达。应用SMARTRACE技术获得GYLZ RCC18基因的全长 ,并证明GYLZ RCC18是一个 5′端有D3种不同剪切方式亚型的基因家族。 结论 GYLZ RCC18基因是肾癌特异表达的新基因。人肾癌cDNA消减文库可作为筛选、克隆肾癌特异性表达的未知新基因的基础。

Objective[WT5”BZ] To clone and identify RCC specially expressed genes different with normal kidney tissue. [WT5”HZ]Methods[WT5”BZ] A technique called suppression subtractive hybridization was used to construct the library which contains the differently expressing cDNAs between RCC and normal kidney. Then the RCC specially expressed genes were cloned from it. [WT5”HZ]Results[WT5”BZ] Human RCC subtractive library with high subtractive efficiency was set up successfully. The amplified library contains 350 positive clones.Sequence analysis was performed in 5 clones. All of the sequences were unknown before and the cDNA inserting GYLZ RCC18 had three copies. Northern blot analysis showed that GYLZ RCC18 cDNA expressed highly in RCC, but there was no any signal could be detected in normal kidney.Using SMART RACE technique,we obtained the full length of novel gene of GYLZ RCC18.We also identified that GYLZ RCC18 family contains 3 subtype genes. [WT5”HZ]Conclusions[WT5”BZ] The highly efficient cDNA subtractive library lays a solid foundation for screening and cloning new and specific oncogenes or tumor suppressor genes of RCC.The novel differentially expressed genes provide an important clue to studying the mechanism of the occurrence and develpment of RCC. [WT5”HZ]