关键词: TIP30基因, 基因转染, 抑瘤作用

Abstract: Abstract:Objective To explore the effect of TIP30 gene on the growth inhibition of renal carcinoma cell line 786-0 and look for a potential therapeutic target for renal carcinoma.Methods TIP30 gene was amplified by reverse transcription-polymerase chain reaction(RT-PCR).A eukaryotic expression vector pcDNA3.1-TIP30 was constructed and transfected into 786-0 cells; pcDNA3.1(+)was also transfected as control.After transfection,the expression of TIP30 in 786-0 cells was detected by RT-PCR and Western blotting.The changes of cell proliferation and cell cycle were observed by MTT and FCM assay.Results The mRNA and protein expressions of TIP30 gene in pcDNA3.1-TIP30-transfected 786-0 cells were significantly increased than those in untreated and pcDNA3.1(+)-transfected cells(P<0.05).However,there was no difference in TIP30 expression between untreated and pcDNA3.1(+)-transfected 786-0 cells(P>0.05).The inhibitory rate of pcDNA3.1-TIP30-transfected 786-0 cells was significantly higher than those in untreated and pcDNA3.1(+)-transfected cells(P<0.01); Cell cycle analysis by flow cytometry showed that the number of cells in G₀-G₁ phase of pcDNA3.1-TIP30-transfected 786-0 cells was significantly increased while the number of cells in phase S and G₂-M was decreased compared with untreated and pcDNA3.1(+)-transfected cells(P<0.01).Conclusion 786-0 cells can stably express exogene TIP30. Transduction of TIP30 gene into lower expression renal carcinoma cells can restore its suppressive effect on cell growth,suggesting that TIP30 gene may be a new therapeutic target for renal carcinoma.

Key words: TIP30 gene, gene transfection, anti-tumor effect