抑癌基因TIP30对肾癌细胞系786-0的生长抑制作用  

Inhibitory effect of TIP30 gene on growth of human renal carcinoma786-0 cells

作　　者：胡敬海[1] 沈彬[1] 徐宏[2] 王春喜[1] 

机构地区：[1]吉林大学第一医院泌尿外科,吉林长春130021 [2]吉林大学第一医院药剂科,吉林长春130021

出　　处：《吉林大学学报（医学版）》2009年第1期87-91,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划项目资助课题(990574-2)

摘　　要：目的:筛选含有外源性TIP30的人肾透明细胞腺癌细胞系786-0,探讨TIP30基因对786-0的生长抑制作用,寻找肾癌基因治疗的潜在靶点。方法:采用RT-PCR技术扩增TIP30基因,构建真核表达载体pcDNA3.1-TIP30,转染786-0细胞,利用RT-PCR及Western blotting检测稳定转染pcDNA3.1-TIP30载体、转染pcDNA3.1(+)空载体及未处理的786-0细胞中TIP30的表达,另通过MTT法检测转染后细胞增殖能力的变化,流式细胞仪测定细胞周期分布的变化。结果:与未处理的和转染pcDNA3.1(+)的786-0细胞比较,转染pcDNA3.1-TIP30的786-0细胞中TIP30基因的mRNA和蛋白表达水平均明显增加(P<0.05);未处理与转染pcDNA3.1(+)的786-0细胞中TIP30的表达水平比较差异无显著性(P>0.05)。转染pcDNA3.1-TIP30的786-0细胞生长抑制率明显高于未处理的和转染pcDNA3.1(+)的786-0细胞(P<0.01)。流式仪检测细胞周期,与未处理的和转染pcDNA3.1(+)的786-0细胞比较,转染pcDNA3.1-TIP30的786-0细胞G0-G1期细胞比例显著增加,S和G2-M期细胞比例降低(P<0.01)。结论:786-0细胞能够稳定表达外源基因TIP30;提高肾癌细胞中TIP30蛋白的表达,可抑制肿瘤细胞的生长;TIP30是肾癌基因治疗的潜在靶点。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 peDNA3.1-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.

关 键 词：肾肿瘤 TIP30基因 基因转染 抑瘤作用 

分 类 号：Q78[生物学—分子生物学] R737.11[医药卫生—肿瘤]