肾癌差异表达基因GYLZ-RCC18的全长克隆及意义

目的 克隆并鉴定肾癌与下沉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的肾癌组织中有明显表达,而在正常肾组织中无表达,应用ＳＭＡＲＴ ＲＡＣＥ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ｃ

Cloning and identifying of renal cell carcinoma differentially expressed genes

OBJECTIVE: To clone and identify RCC specially expressed genes different with normal kidney tissue. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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 development of RCC.