| miR-130a-3p 通过调控 PDK1 影响肝癌细胞糖代谢的研究 |
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| 引用本文： | 李海龙,石杰,郭林池,戚琪,于晓光. miR-130a-3p 通过调控 PDK1 影响肝癌细胞糖代谢的研究[J]. 医学分子生物学杂志, 2022, 19(3): 206-212. DOI: 10.3870/j.issn.1672-8009.2022.03.005 |
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| 作者姓名： | 李海龙 石杰 郭林池 戚琪 于晓光 |
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| 作者单位： | 1哈尔滨医科大学基础医学院生物化学与分子生物学教研室 哈尔滨市, 150081 2宁夏回族自治区人民医院全科医学 银川市, 750001 |
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| 基金项目： | 宁夏回族自治区自然科学基金 |
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| 摘 要： | 目的 探讨 miR-130a-3p 通过靶向糖代谢关键酶 PDK1 对肝癌细胞糖代谢以及增殖迁移的影响,为肝癌的诊断及治疗提供新的方向。 方法 qRT-PCR 技术检测肝癌的临床组织和细胞中 miR-130a-3p 的表达, 克隆形成、 CCK-8、 Transwell 和细胞划痕实验检测细胞的增殖和迁移; Western 印迹检测相关蛋白的表达; 乳酸检测试剂盒, ATP 检测试剂盒和 PDH 活性检测试剂盒分别检测细胞中乳酸的生成、 ATP 的生成和PDH 的活性; 双荧光素酶报告基因实验验证 miR-130a-3p 与 PDK1 3′UTR 靶向结合。 结果 在肝癌组织和细胞中 miR-130a-3p 呈低表达, 并与患者肿瘤组织的大小和 TNM 分期有关; 过表达 miR-130a-3p 能够增强肝癌细胞中 PDH 活性, 抑制乳酸和 ATP 的生成, 从而抑制肝癌细胞的增殖和迁移; miR-130a-3p 可以靶向调控 PDK1; 抑制 PDK1 的表达, 能够逆转 miR-130a-3p 过表达对细胞增殖和迁移能力的增强, 此外对 PDH 活性、 乳酸和 ATP 的影响也被逆转。 结论 miR-130a-3p 通过靶向调控 PDK1 影响肝癌细胞糖酵解及恶性进展。
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| 关 键 词： | miR-130a-3p PDK1 肝癌 糖代谢 |
Effect of miR-130 a-3 p on Glucose Metabolism of Liver Cancer Cells by Regulating PDK1 |
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| LI Hailong,SHI Jie,GUO Linchi,QI Qi,YU Xiaoguang. Effect of miR-130 a-3 p on Glucose Metabolism of Liver Cancer Cells by Regulating PDK1[J]. Journal of Medical Molecular Biology, 2022, 19(3): 206-212. DOI: 10.3870/j.issn.1672-8009.2022.03.005 |
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| Authors: | LI Hailong SHI Jie GUO Linchi QI Qi YU Xiaoguang |
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| Affiliation: | 1Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin, 150081,China 2Department of General Practice, People’s Hospital of Ningxia Hui Autonomous Region, Yinchuan,750001, China |
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| Abstract: | Objective To investigate the effect of miR-130a-3p on glucose metabolism, proliferation and migration of liver cancer cells by targeting PDK1, a key enzyme of glucose metabolism,so as to provide a new target for the diagnosis and treatment of liver cancer. Methods The expression of miR-130a-3p in clinical tissue samples and cell lines of hepatocellular carcinoma was detected by qRT-PCR. Cell proliferation was measured by clone formation and CCK-8. Cell migration wasmeasured by transwell and wound healing assay. Western blotting was used to detect the expression ofrelated proteins. The production of lactic acid and ATP in cells was assayed by lactate assay kit andATP assay kit respectively, and the PDH activity in cells was assayed by PDH activity assay kit. Thedual-luciferase reporter gene assay was used to verified the binding of miR-130a-3p to PDK1 3′UTR. Results The expression level of miR-130a-3p was lower in liver cancer tissues and cells,and it was related to the size of tumor tissues and TNM stage. Overexpression of miR-130a-3p couldenhance the activity of PDH in hepatoma cells, reduce the production of lactic acid and ATP, andinhibit the proliferation and migration of hepatoma cells. miR-130a-3p could bind to PDK1 and regulate its expression. Inhibition of PDK1 expression could reverse the enhancing effects of miR-130a-3poverexpression on cell proliferation and migration, and the effects on PDH activity, production oflactic acid and ATP were also reversed. Conclusion miR-130a-3p affects glycolysis and malignantprogression of hepatoma cells by targeting PDK1. |
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| Keywords: | miR-130a-3p PDK1 liver cancer glucose metabolism |
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