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

发布时间：2018-11-04 20:07

【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。
[Abstract]:Aim: to study the biological effects and transcriptional regulation of amino acid transporter gene SLC7A5 (solute carrier family 7 member 5 in tumor cells. Methods: the expression of SLC7A5 in human normal tissues and corresponding tumor tissues was analyzed by using gene expression and (Gene Expression Omnibus,GEO database. The recombinant plasmid of SLC7A5 gene was constructed and the effect of the gene on the proliferation of tumor cells was detected by methylthiazolyldiphenyl-tetrazolium bromide,MTT colorimetry and flow cytometry. The promoter plasmid of SLC7A5 gene and its transcription factor binding site were constructed by bioinformatics, and the luciferase reporter gene system was used. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to investigate the regulation of transforming growth factor 尾 1 (transforming growth factor- 尾 1 TGF- 尾 1 on the expression of SLC7A5 gene. Results: GEO database analysis showed that the distribution of SLC7A5 was tissue specific and the expression level in most tumor tissues was significantly higher than that in normal tissues. The results of MTT colorimetry and flow cytometry showed that SLC7A5 could promote cell proliferation. Through promoter analysis, vector construction, reporter gene detection and RT-PCR assay, it was confirmed that TGF- 尾 1 could up-regulate the activity of SLC7A5 promoter and promote the expression of the gene. Conclusion: SLC7A5 gene plays a promoting role in tumor cells and is regulated by TGF- 尾 1 signaling pathway.
【作者单位】：湖南大学生物学院;
【基金】：国家自然科学基金(81270735)~~
【分类号】：R730.2

【相似文献】