Types And Expression Patterns Of Alternatively Spliced Variants Of TCF-19 In Clear Cell Renal Cell Carcinoma

[Objective]To study the existance and expression of alternative spliced isoforms of TCF19, a cell Growth regulatory gene, in human clear cell renal cell carcinoma (ccRCC) tissues and their relationship to the tumor.[Method]cDNA microarray technique was used to Survey the differentially expressed genes between the two ccRCC cell lines (RCC05-TXJ and RCC05-ZYJ). After screening with DNA microarray, Choose TCF19 gene to be studied. Alternative Splicing Database was used to predict the splice variants of TCF19. According to the prediction,Designing the RT-PCR primers of TCF19 gene by using Primer Premier 5.0 software, Oligo 6.0 software and BLAST program. Using the specific primer sets, the splice variants were identified in primary ccRCCs and normal tissues adjacent to cancer. The variants were amplified by RT-PCR, and were cloned into pMD18-T vector, which were subsequently transformed into E.Coli DH5a. After affirmation the existence of variants by gene sequencing, the mRNA expression of TCF19-1,TCF19-3 were detected in cancer tissues, and normal tissues side to cancer,then designing the realtime primers of TCF19-1,TCF19-3,To study the expression difference of two kinds of isomers in cancer tissues and normal tissue side to cancer by realtime.The tissue specimens were obtained from the Urology,which Affiliated Hospitals of Second Military Medical University from Jan.2005 to Dec.2008, Tissue specimens from 32 patients (ages from 36-78) with ccRCC were collected after surgical treatments immediately. All specimens were saved in liquid nitrogen and stored at-80℃.[Result]Two isoforms of TCF-19 (TCF19-1,TCF19-3)was identified from 5 predicteed isomers, And TCF19-1 express in cancer tissue (26/32) higher than that in normal tissue side to cancer (6/32). the expression of TCF19-3 was not significantly different yet. The results revealed that the transcripts of TCF19-1 was related with the occurrence and progress of primary ccRCC likely. (P<0.05),[Conclution] Two new isoforms of TCF19 (TCF19-1 and TCF19-3) were found in ccRCC and the adjacent tissues. Expression of TCF19-1 was significantly higher in primary ccRCCs than in the adjacent normal tissues (P< 0.05). No significant difference in the expression of TCF19-3 in the primary tumor and adjacent tissues was found (P=0.082).