国际肿瘤学杂志

国际肿瘤学杂志 ›› 2017, Vol. 44 ›› Issue (11): 819-823.doi: 10.3760/cma.j.issn.1673-422X.2017.11.004

• 论著 • 上一篇    下一篇

miR-320通过靶向E2F1基因抑制结直肠癌细胞的糖代谢

刘延锋，王铎，张华洲，张允东   

  1. 710038西安医学院第二附属医院普外科
  • 收稿日期:2017-07-04 出版日期:2017-11-08 发布日期:2017-11-24
  • 通讯作者: 刘延锋 E-mail:liliyan204@163.com

miR-320 inhibits glycometabolism in colorectal cancer by targeting E2F1 gene

Liu Yanfeng, Wang Duo, Zhang Huazhou, Zhang Yundong   

  1. Department of General Surgery, Second Affiliated Hospital of Xi′an Medical University, Xi′an 710038, China
  • Received:2017-07-04 Online:2017-11-08 Published:2017-11-24
  • Contact: Liu Yanfeng E-mail:liliyan204@163.com

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摘要： 目的研究微小RNA（miR320）靶向E2F1基因对结直肠癌的肿瘤糖代谢的影响。方法使用实时荧光定量聚合酶链反应（qRTPCR）检测miR320在结直肠癌细胞株和癌组织中的表达水平。生物信息学预测miR320与E2F1的结合位点。荧光素酶实验检测miR320是否可靶向调节E2F1。从mRNA水平及蛋白水平验证E2F1与miR320相互关系，使用葡萄糖/葡萄糖氧化酶法试剂盒和乳酸检测试剂盒分析在SW480和LOVO细胞中过表达miR320及干扰E2F1表达后，肿瘤细胞糖代谢的变化。结果qRTPCR测得miR320在结直肠癌细胞株（F=42.327，P＜0.001）和癌组织（t=4.345，P=0.023）中低表达，荧光素酶报告基因检测miR320可靶向负调节E2F1的表达（t=4.716，P=0.042），mRNA水平（t=4.780，P=0.041；t=5.506，P=0.031）和蛋白水平证实在LOVO和SW480细胞株中E2F1与miR320可相互作用，在SW480和LOVO中过表达miR320可使细胞上清中的葡萄糖（t=5.262，P=0.034；t=21.079，P=0.002）和乳酸（t=9.609，P=0.011；t=18.582，P=0.003）含量降低，同时降低E2F1的表达，则可加强miR320对葡萄糖（t=5.128，P=0.036；t=5.089，P=0.037）和乳酸（t=8.573，P=0.013；t=13.364，P=0.006）含量的抑制作用。结论E2F1是miR320的靶基因，miR320通过靶向E2F1基因调节结直肠癌细胞的肿瘤糖代谢。

关键词: 结直肠肿瘤, 微RNAs, 代谢, E2F1转录因子

Abstract: ObjectiveTo study the effect of microRNA320 （miR320） targeting E2F1 gene on tumor glycometabolism in colorectal cancer. MethodsThe miR320 expression level in colorectal cancer cell lines and cancer tissues was detected using quantitative realtime polymerase chain reaction (qRTPCR). The binding sites of miR320 and E2F1 were predicted by bioinformatics. Luciferase assay was used to detect the targeting regulation of miR320 on E2F1. The relationship between E2F1 and miR320 was verified in mRNA level and protein level. When the miR320 in SW480 and LOVO cells was upregulated and the E2F1 was downregulated, the changes of glycometabolism in tumor cells were analyzed using glucose/glucose oxidase kit and lactate test kit. ResultsThe qRTPCR results showed low expressions of miR320 in colorectal cancer cell lines and cancer tissues (F=42.327, P＜0.001; t=4.345, P=0.023). Luciferase assay showed that miR320 could negatively regulate the expression of E2F1 (t=4.716, P=0.042). The expression levels of E2F1 protein and mRNA (t=4.780, P=0.041; t=5.506, P=0.031) confirmed that miR320 could interact with E2F1 in LOVO and SW480 cells. Overexpression of miR320 could reduce the contents of glucose (t=5.262, P=0.034; t=21.079, P=0.002) and lactic acid (t=9.609, P=0.011; t=18.582, P=0.003) in the cellular supernatant in SW480 and LOVO cells. Downregulating the expression of E2F1 at the same time could enhance the inhibitory effect of miR320 on glucose (t=5.128, P=0.036; t=5.089, P=0.037) and lactic acid (t=8.573, P=0.013; t=13.364, P=0.006). ConclusionE2F1 is the target gene of miR320, and miR320 can regulate the glycometabolism of colorectal cancer cells by targeting E2F1 gene.