普罗布考通过调节肝癌细胞HepG2代谢重编程逆转Warburg效应抑制其转移和侵袭的机制研究

Probucol inhibited metastasis and invasion by reversing Warburg effect and regulated reprogramming of hepatoma cell HepG2

目的:研究普罗布考调节HepG2的代谢重编程逆转Warburg效应抑制其转移和侵袭的机制,为脂代谢调节药物的研究提供支持。方法:HepG2细胞体外培养后分为对照组(NC)、高剂量组(High)、中剂量组(Middle)和低剂量组(Low),其中高、中、低剂量组的细胞分别使用100、50、10μmol/L的普罗布考干预。CCK-8检测细胞活力,流式细胞术检测细胞周期的变化,细胞划痕实验检测细胞转移能力,Transwell小室实验检测细胞侵袭能力,Western blot法检测细胞中己糖激酶(HK)、丙酮酸激酶(PK)、乳酸脱氢酶(LDH)、磷酸果糖激酶(PFKP)、丙酮酸脱氢酶(PDH)以及柠檬酸合酶(CS)的表达变化,试剂盒检测培养基中乳酸的含量,ECAR检测细胞糖酵解的压力,929 oxygen system检测细胞对于氧气的吸收水平检测。结果:普罗布考可以抑制HepG2的增殖,且有剂量依赖,还可以阻滞细胞周期。普罗布考可以提高HK、PK、LDH、PFKP以及PDH和CS的表达,增加耗氧量减少乳酸的产生,促进细胞的三羧酸循环。普罗布考还可以抑制HepG2的侵袭和迁移能力。结论:普罗布考可以调节肝癌细胞HepG2的代谢重编程、逆转Warburg效应从而抑制肿瘤细胞的增殖、迁移和侵袭。

AIM： To study the mechanism of probucol-modulated the metabolic reprogramming of HepG2,reversed the Warburg effect and inhibited its metastasis and invasion,so as to provide support for the research of lipid metabolism regulating drugs.METHODS： HepG2 cells were divided into control group（ NC）,high dose group（ High）,middle dose group（ Middle） and low dose group（ Low） after cultured in vitro. The cells in the high dose,middle dose and low dose group were treated with probucol of 100,50,10 μmol/L,respectively. CCK-8 was used to detect cell viability; the cell cycle changes were detected by flow cytometry; the cell migration assay was performed by cell scratch test; the cell invasion ability was detected by Transwell experiment;the expression of hexokinase（ HK）,pyruvate kinase（ PK）,lactate dehydrogenase（ LDH）,phospho fructose kinase（ PFKP）,pyruvate dehydrogenase（ PDH） and citric acid synthase（ CS） in cells were detected by Western blot; the level of lactic acid in the medium was detected by the kit; the pressure of glycolysis was detected by ECAR; and the oxygen absorption was detected by the 929 oxygen system.RESULTS： Probucol dose-dependently inhibited the proliferation of HepG2,and blocked cell cycle. Probucol increased the expression of HK,PK,LDH,PFKP,PDH and CS,increased oxygen consumption and reduced the production of lactic acid,and promoted the cycle of three carboxylic acids. Probucol also inhibited the invasion and migration of HepG2.CONCLUSION： Probucol can regulate the metabolic reprogramming,reverse the Warburg effect of hepatoma cell HepG2,which can inhibit the proliferation,migration and invasion of tumor cells.