Using Suppression Subtractive Hybridization Technique To Clone Differentially Expressed Genes Between Renal Cell Carcinoma And The Matched Normal Renal Epithelium

Renal cell carcinoma(RCC) is of great malignance.Itsclinical manifestation is multiform.It aceouts forapproximately 3% of adult malignaneies.The ineidenee of RCCinereases about 2% all over the world every year.Nearly 100,000patients died from RCC every year.In China,the ineidence ofRCC is on the second place of urologie tumors and the ineidencehas increased in recent years.It is a great threat to humanhealth.There is a large number of genes related to RCC incontrolling cell growth,cell differentiation and cell death.order to find out the initiation and progression mechanism RCC It is necessary to identify and elone the RCCdifferentially expressed genes and find out the RCC geneprofile.This is of great value to the RCC diagnosis,biologiealtreatment and prognosis analysis. There are several teehniques used to clone differentiallyexpressed genes.Suppression subtraetive hybridization(SSH)technique established by Diatehenko has more merits than theother ones.In an attempt to identify genes differentiallyexpressed in RCC we used the method of suppression subtraetivehybridization to eonstruet an RCC cDNA subtraetivebetween human RCC tissue and normal renal epithelium.Then thedifferentially expressed genes were seleeted by using dotblotting.Some of the Positive elones were sequeneed randomly.The result was analyzed with the Blast 2.0 softwere in GenBank. The results showed that human RCC cDNA subtraetive1ibrary with high subtraetive effieieney was set upsueeessfully.The library ineluded 400 positive elones.Twohundred elones were seleeted randomly to be amPlified.Onehundred and seventy seven elones were real Positive under dotblotting analysis.Fifty five elones with inserted fragmentswere sequeneed.The blast result of GenBank showed that therewere 26 kinds of genes.Twenty four kinds of genes were known,and two eDNA elones did not mateh with any sequenees in theGenBank,indieating that they may be novel genes. With the human RCC subtractive library set upsueeessfully we obtained not only some known genes related totumor but also some novel genes.The known genes we obtainedval idated the Previous researeh results while the novel geneslaid foundation for analyzing the new genes'fuetion.Thesematerials Provides an1 nstruet for sPeeifie diagnosis,biologietheraPy and Prognosis analysis in future.That the human RCCsubtraetive library was establied sueeessfully showed SSHtechnique 15 an effieient method to identify and elone RCCdifferentially exPressed genes.Thislarge seale sereening and eloning of newmakes it Possible for and sPeeifie RCC genes In future.More over it providesanew1 nitiation and develoPment meehanism method for studying the of RCC.