SLC7A5对肿瘤细胞增殖的影响及其与转化生长因子-β1信号调控的关系  

Effect of SLC7A5 on the proliferation of tumor cells and its relationship with transforming growth factor-β1 signal pathway

作　　者：赵婷[1] 李丹[1] 刘晓雯[1] 刘智勇[1] 吴道兵[1] 周世华[1] 段红艳[1] 

机构地区：[1]湖南大学生物学院,长沙410082

出　　处：《中南大学学报（医学版）》2017年第5期485-492,共8页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(81270735)~~

摘　　要：目的:研究氨基酸转运载体基因SLC7A5(solute carrier family 7,member 5)在肿瘤细胞中发挥的生物学效应及其转录调控机制。方法:利用基因表达综合(Gene Expression Omnibus,GEO)数据库分析SLC7A5在人正常组织和相应的肿瘤组织中的表达情况;构建SLC7A5基因的重组质粒,采用噻唑蓝(methylthiazolyldiphenyl-tetrazolium bromide,MTT)比色法、流式细胞仪技术检测该基因对肿瘤细胞增殖的影响;通过生物信息学方法分析SLC7A5基因启动子及其转录因子结合位点,构建该基因的启动子质粒,并利用荧光素酶报告基因系统、反转录聚合酶链反应(RT-PCR)探讨转化生长因子-β1(transforming growth factor-β1,TGF-β1)对SLC7A5基因的表达调控。结果:GEO数据库分析显示SLC7A5的分布具有组织特异性,且在大多数的肿瘤组织中的表达水平明显高于相应的正常组织;MTT比色法、流式细胞仪检测结果表明SLC7A5具有促进细胞增殖的作用;通过启动子分析、载体构建、报告基因检测和RT-PCR实验证实TGF-β1可上调SLC7A5启动子的活性,从而促进该基因的表达。结论:SLC7A5基因在肿瘤细胞中发挥促癌作用,且受TGF-β1信号通路的调控。Objective： To explore the biological effects of amino acid transporter gene SLC7A5 （solute carrier family 7, member 5） on tumor cells and the regulatory mechanism at transcriptional level. Methods： Tae expression of SLC7A5 was examined in human normal tissues and corresponding tumor tissues by Gene Expression Omnibus （GEO） database. The recombinant plasmid of SLC7A5 gene was constructed, and the effect of the SLC7A5 gene on tumor cell proliferation was investigated by methylthiazolyldiphenyl-tetrazolium bromide （MTT） and flow cytometry. SLCTA5 gene promoter and transcription factor binding sites were predicted through bioinformatics analysis, and the gene promoter recombinant plasmid was constructed. Then the dual luciferase reporter gene assay and reverse transcription polymerase chain reaction （RT-PCR） were used to explore the regulation of transforming growth factor-β1（TGF-β1） signal on SLC7A5 gene expression. Results： The GEO database analysis showed that the distribution of SLC7A5 was tissue specific, and its expression level was significantly higher in the tumor tissues than that in the corresponding normal tissues. The results of MTT and flow cytometry showed that SLC7A5 could promote cell proliferation. Results from the promoter analysis, reporter gene assay and RT-PCR confirmed that TGF-β1 could up-regulate the activity of SLC7A5 promoter and promote the expression of the SLC7A5 gene. Conclusion： SLC7A5 gene plays a role in promoting tumor development, which is regulated by the TGF-β1 signaling pathway.

关 键 词：SLC7A5基因 细胞增殖 TGF-β1信号通路 

分 类 号：R730.2[医药卫生—肿瘤]