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[Cloning and identifying of renal cell carcinoma differentially expressed genes]
1Department of Urology, First Hospital, Peking University, Beijing 100034, China.
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.