干扰骨桥蛋白表达对肝癌细胞Warburg效应的影响及机制
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内蒙古医科大学

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Effect of osteopontin (OPN) interference on Warburg effect ofhepatoma cells and its mechanism
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Inner Mongolia Medical University

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    摘要:

    目的 探讨小干扰RNA(siRNA)靶向抑制骨桥蛋白(OPN)表达后对肝癌细胞Warburg效应的影响及其作用机制。方法 将靶向OPN的siRNA(si-OPN)及阴性对照序列(si-NC)分别转染肝癌细胞HepG2和SK-HEP-1。采用2-NBDG荧光探针法检测肝癌细胞的葡萄糖摄取能力,乳酸试剂盒检测肝癌细胞乳酸生成情况。采用RT-qPCR及Western blot检测肝癌细胞中糖酵解相关基因GLUT1、HK2及LDHA的表达变化。在HepG2和SK-HEP-1细胞中沉默LGALS3BP表达后,Western blot分别检测肝癌细胞中LGALS3BP及糖酵解相关基因GLUT1、HK2及LDHA的表达变化,2-NBDG法检测细胞葡萄糖摄取能力和乳酸试剂盒检测乳酸生成情况。 结果 与未转染组(control)组和si-NC组相比,干扰OPN组(si-OPN)组的HepG2和SK-HEP-1细胞对葡萄糖的摄取量及乳酸生成量均显著降低,同时GLUT1、HK2及LDHA基因在mRNA及蛋白水平的表达均显著降低。HepG2和SK-HEP-1细胞中干扰OPN后LGALS3BP的蛋白表达下调,干扰LGALS3BP后葡萄糖摄取量和乳酸产量的水平显著降低,GLUT1、HK2及LDHA的蛋白表达显著降低。结论 干扰OPN能通过下调LGALS3BP抑制糖酵解相关基因的表达,从而抑制肝癌细胞的Warburg效应。

    Abstract:

    Objective To investigate the effect of osteopontin (OPN) interference on the Warburg effect of hepatoma cells and the related molecular mechanism. Methods Small interfere RNA (siRNA) targeting OPN (si-OPN) and negative control (si-NC) were transfected into HepG2 and SK-HEP-1 cells. 2-NBDG fluorescent probe was used to detect the glucose uptake capacity of hepatoma, and the lactate detection kit was used to evaluate the lactate production in HepG2 and SK-HEP-1 cells. The expression of glycolysis-related genes, suchas GLUT1, HK2 and LDHA was assessed by RT-qPCR and western blot assays. After silencing the expression of LGALS3BP in HepG2 and SK-HEP-1 cells, Western blot was used to detect the expression of LGALS3BP, GLUT1, HK2 and LDHA. The glucose uptake and lactate production were also evaluated in LGALS3BP silenced HepG2 and SK-HEP-1 cells. Results Compared with the control group and si-NC group, the levels of glucose uptake and lactate production were significantly reduced in HepG2 and SK-HEP-1 cells interfe

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  • 收稿日期:2025-01-14
  • 最后修改日期:2025-07-14
  • 录用日期:2025-09-10
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