肾癌差异表达基因克隆的研究

目的 克隆并鉴定肾癌与正常肾之间差异表达的基因。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差异表达的基因。结果 构建成功了高消减效率的人肾癌cDNA消减文库，对其中5个克隆插入的cDNA片段进行测序后经Genbank检索表明，5个片段均为未知新序列，其中GYIZ-RCC18为3个拷贝，提示以上5个cDNA征段可能来自3个新基因。Northern杂交分析显示，GYIZ-RCC18在肾癌组织中有明显表达，而在正常肾组织中无表达，这证明GYIZ-RCC18是肾癌相关的新基因。结论 人肾癌cDNA消减文库的建立为进一步筛选，克隆肾癌特异性表达的新基因奠定了基础。

Cloning and identifying renal cell carcinoma differentially expressed genes and their significance

Objective To Clone study the differentially expressed new genes in renal cell carcinoma (RCC). Methods Using a technique known as suppression subtractive hybridization to construct the library which contains the differently expressing cDNAs between RCC and normal kidney cells. Then the RCC specifically expressed genes were cloned. Results Human RCC subtrctive library with high subtractive efficiency was set up sucessfully. The amplified library contained 350 positive clones. Sequence analysis were performed for 5 clones. All the sequences were unknown previioously and the cDNA insert GYLZ RCC18 had three copie. Northern blot analysis showed that GYLZ RCC18 cDNA expressed highly in RCC, but no signal could be detected in normal kidney. Using SMART RACE technique, the full length of novel gene of GYLZ RCC18 was obtained. Conclusions The highly efficient cDNA subtractive library may have formed solid foundation for screening and cloning new and specific oncogenes or tumor suppressor genes of RCC. The novel differentially expressed genes may provide an important clue for studying the mechanism of occurrence and develpment of RCC.