摘要
目的:探讨转录因子SREBP1通过促进SLC16A8表达调控肿瘤酸性微环境参与结直肠癌(CRC)发生发展的机制。方法:通过生物信息学数据库分析SLC16A8在CRC中的表达情况及其与患者预后的关系。使用RT-qPCR法检测并比较各CRC细胞系中SLC16A8的表达水平;过表达SLC16A8后使用CCK-8法检测细胞增殖活性;使用Transwell法检测细胞侵袭能力;使用Metascape数据库对SLC16A8的功能富集进行分析;使用乳酸试剂盒检测细胞上清液中乳酸水平。通过生物信息学数据库分析SLC16A8的转录因子及潜在结合位点;通过双荧光素酶实验检测SREBP1蛋白与SLC16A8相关性;通过UALCAN portal分析SREBP1 mRNA表达水平;通过Kaplan-Meier Plotter分析CRC中SREBP1表达水平与预后的关系;通过GEPIA2分析SREBP1表达水平与SLC16A8的相关性;使用Western blot分析过表达SREBP1对SLC16A8蛋白表达的影响。通过拯救实验分析SREBP1是否通过正调控SLC16A8促进乳酸转运。结果:SLC16A8高表达的CRC患者提示预后较差(P<0.05),且SLC16A8在CRC组织及细胞系中均为高表达水平(P<0.05)。过表达SLC16A8可提高CRC细胞增殖及侵袭能力。功能富集分析数据表明SLC16A8的功能聚集在乳酸转运及血管生成,且SLC16A8可提高CRC细胞上清液中乳酸水平。双荧光素酶实验证实SREBP1可与SLC16A8直接结合。SREBP1在CRC组织中高表达并与患者预后较差具有相关性(P<0.05)。SREBP1可促进SLC16A8蛋白表达,且SREBP1可通过正调控SLC16A8促进乳酸转运。结论:SLC16A8可能通过调控肿瘤酸性微环境促进CRC细胞增殖和侵袭,转录因子SREBP1通过上调SLC16A8表达参与CRC发生发展。
Objective:To study the molecular mechanism that transcription factor SREBP1 involves in the process of colorectal cancer(CRC)by promoting the expression of SLC16A8 and regulating acidic tumor microenvironment.Methods:The expression of SLC16A8 in CRC and the association between SLC16A8 on poor prognosis were analyzed by bioinformatics database.The expression of SLC16A8 in CRC cells were detected by quantitative real-time reverse-transcription(RT-qPCR).CCK-8 was used to analyze cell proliferation,and Transwell was used to analyze cell invasive ability.A functional enrichment analysis was carried out with Metascape database.The levels of lactic acid in supernatant was monitored with lactate kit.Bioinformatics method was used to screen transcription factor which could bind to SLC16A8 and potential transcription factor binding sites,and dual luciferase reporter assay was utilized to the correlation between SREBP1 protein and SLC16A8.UALCAN portal was used to analysis SREBP1 mRNA expression,relationship between the expression of SREBP1 and prognosis was analyzed by Kaplan-Meier Plotter.GEPIA2 was used to analysis the relationship between the expression of SREBP1 and SLC16A8.The effect of overexpression of SREBP1 on the expression of SLC16A8 protein were analyzed by Western blot.Rescue experiments were used to analyse whether SREBP1 promotes lactate transport through positive regulating SLC16A8.Results:CRC patients with high expression of SLC16A8 indicate poor prognosis(P<0.05),and SLC16A8 was highly expressed in CRC tissues and cell lines(P<0.05).Overexpression of SLC16A8 enhanced cell proliferation and invasion.GO enrichment analysis data further indicated that its functions were involved in the lactic acid transport and angiogenesis,and SLC16A8 could promote the levels of lactic acid in supernatant.The double Luciferase test confirmed that SREBP1 could bind directly to SLC16A8.SREBP1 was highly expressed in CRC tissue and was correlated with poor patient prognosis(P<0.05)SREBP1 could promote the expression of SLC16A8
作者
彭明沙
周翼
刘刚
冯雪雅
彭洪
PENG Ming-sha;ZHOU Yi;LIU Gang;FENG Xue-ya;PENG Hong(Department of Anorectal Surgery,the Second Clinical Medical College of North Sichuan Medical College,Nanchong 637000,Sichuan,China;Department of Ultrasound,Nanchong Central Hospital,the Second Clinical Medical College of North Sichuan Medical College,Nanchong 637000,Sichuan,China)
出处
《川北医学院学报》
CAS
2023年第6期729-735,共7页
Journal of North Sichuan Medical College
基金
四川省南充市市校合作项目(22SXQT0069,22SXQT0070,22SXQT0063)
川北医学院校级项目(CBY23-QA-Y53)
四川省青年创新科研项目(Q22049)。
