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

doi:10.12092/j.issn.1009-2501.2018.06.001

Abstract

Abstract:

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.

Key words: probucol, metabolic reprogramming, Warburg effect, invasion

CLC Number:

R965.2

WANG Jin, YANG Yi, GUO Li, LI Wenyan, RUAN Shuiliang. Probucol inhibited metastasis and invasion by reversing Warburg effect and regulated reprogramming of hepatoma cell HepG2[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(6): 601-607.

References

［1］Wei GX, Wang SL, Zhang L, et al. Impact of postoperative pituitary tumor adrenal insufficiency and hormone replacement therapy (HRT) on tumor recurrence［J］. Eur Rev Med Pharmacol Sci, 2017:2671-2675.
［2］Liu J, Zhang C, Hu W, et al. Tumor suppressor p53 and its mutants in cancer metabolism［J］. Can Lett, 2015, 356(2):197-203.
［3］MerinoRamos T, VzquezCalvo , Casas J, et al. Modification of the host cell lipid metabolism induced by hypolipidemic drugs targeting the acetyl coenzyme a carboxylase impairs west Nile virus replication［J］. Antimicrob Agents Chemother, 2015, 60(1):307-315.
［4］李婷, 胡夕春. 非诺贝特抗肿瘤作用的研究进展［J］. 中华乳腺病杂志:电子版, 2013, 7(5):40-44.
［5］Savic LJ, Chapiro J, Duwe G, et al. Targeting glucose metabolism in cancer: new class of agents for locoregional and systemic therapy of liver cancer and beyond ［J］ ? Hepat Oncol, 2016, 3(1):19-28.
［6］Hsu PP, Sabatini DM. Cancer cell metabolism: Warburg and beyond［J］. Cell, 2008, 134(5):703-707.
［7］Wei H, Guo L, Li L, et al. Mechanism of Warburg effect and its effect on tumor metastasis［J］. Chin J Lung Cancer, 2015, 18(3):179-183.
［8］Gordon N, Skinner AM, Pommier RF, et al. Gene expression signatures of breast cancer stem and progenitor cells do not exhibit features of Warburg metabolism［J］. Stem Cell Res Ther, 2015, 6(1):157-162.
［9］Sugiyama Y, Hagiya Y, Nakajima M, et al. The heme precursor 5aminolevulinic acid disrupts the Warburg effect in tumor cells and induces caspasedependent apoptosis［J］. Oncol Rep, 2014, 31(3):1282-1286.
［10］Xu Q, Zhang Q, Ishida Y, et al. EGF induces epithelialmesenchymal transition and cancer stemlike cell properties in human oral cancer cells via promoting Warburg effect［J］. Oncotarget, 2016, 8(6):9557-9571.
［11］Hardie DG, Alessi DR. LKB1 and AMPK and the cancermetabolism link ten years after［J］. BMC Biology, 2013, 11(1):36.
［12］Tanaka T, Ikegami Y, Nakazawa H, et al. Lowdose farnesyltransferase inhibitor suppresses HIF1α and snail expression in triplenegative breast cancer MDAMB231 cells in vitro［J］. J Cell Physiol, 2016, 232(1):192-201.
［13］冯建飞, 穆瑞斌. 他汀类调脂药物联合普罗布考降低血脂的疗效观察［J］. 临床医学研究与实践, 2017, 2(24):37-38.
［14］李燕妮, 仲健, 郭秋野,等. 糖尿病肾病患者接受普罗布考联合奥美沙坦酯治疗后糖脂代谢、氧化和炎症反应程度的评估［J］. 海南医学院学报, 2016, 22(22):2699-2702.
［15］Chen ZP, Liu JH, Jiang DY, et al. Clinical observation of blood sugar uncontrolled nonproliferative diabetic retinopathy patients treated by probucol［J］. Chin J Arterioscl, 2011, 19(9):751-755.

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