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  2. ERK/PKM2 Is Mediated in the Warburg Effect and Cell Proliferation in Arsenic-Induced Human L-02 Hepatocytes

ERK/PKM2 Is Mediated in the Warburg Effect and Cell Proliferation in Arsenic-Induced Human L-02 Hepatocytes

  • Biol Trace Elem Res. 2023 May 27. doi: 10.1007/s12011-023-03706-z.
Fanshuo Yin 1 2 Xin Zhang 1 2 Zaihong Zhang 1 2 Meichen Zhang 1 2 Yunyi Yin 1 2 Yanmei Yang 3 4 Yanhui Gao 5 6
Affiliations

Affiliations

  • 1 Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, No.157 Baojian Road, Nangang District, Harbin, 150081, Heilongjiang Province, China.
  • 2 Key Lab of Etiology and Epidemiology, Education Bureau of Heilongjiang Province & Ministry of Health of P. R. China, Harbin Medical University, Harbin, 150081, Heilongjiang Province, China.
  • 3 Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, No.157 Baojian Road, Nangang District, Harbin, 150081, Heilongjiang Province, China. yangyanmei@hrbmu.edu.cn.
  • 4 Key Lab of Etiology and Epidemiology, Education Bureau of Heilongjiang Province & Ministry of Health of P. R. China, Harbin Medical University, Harbin, 150081, Heilongjiang Province, China. yangyanmei@hrbmu.edu.cn.
  • 5 Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, No.157 Baojian Road, Nangang District, Harbin, 150081, Heilongjiang Province, China. gaoyanhui@hrbmu.edu.cn.
  • 6 Key Lab of Etiology and Epidemiology, Education Bureau of Heilongjiang Province & Ministry of Health of P. R. China, Harbin Medical University, Harbin, 150081, Heilongjiang Province, China. gaoyanhui@hrbmu.edu.cn.
Abstract

This study aimed to investigate the potential role of Pyruvate Kinase M2 (PKM2) and extracellular regulated protein kinase (ERK) in arsenic-induced cell proliferation. L-02 cells were treated with 0.2 and 0.4 μmol/L As3+, glycolysis inhibitor (2-deoxy-D-glucose,2-DG), ERK Inhibitor [1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)-butadiene, U0126] or transfected with PKM2 plasmid. Cell viability, proliferation, lactate acid production, and glucose intake capacity were determined by CCK-8 assay, EdU assay, lactic acid kit and 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]-D-glucose (2-NBDG) uptake kit, respectively. Also, levels of PKM2, phospho-PKM2S37, glucose transporter protein 1 (GLUT1), Lactate Dehydrogenase A (LDHA), ERK, and phospho-ERK were detected using Western blot and the subcellular localization of PKM2 in L-02 cells was detected by immunocytochemistry (ICC). Treatment with 0.2 and 0.4 μmol/L As3+ for 48 h increased the viability and proliferation of L-02 cells, the proportion of 2-NBDG+ cell and lactic acid in the culture medium, and GLUT1, LDHA, PKM2, phospho-PKM2S37, and phospho-ERK levels and PKM2 in nucleus. Compared with the 0.2 μmol/L As3+ treatment group, the lactic acid in the culture medium, cell proliferation and cell viability, and the expression of GLUT1 and LDHA were reduced in the group co-treated with siRNA-PKM2 and arsenic or in the group co-treated with U0126. Moreover, the arsenic-increased phospho-PKM2S37/PKM2 was decreased by U0126. Therefore, ERK/PKM2 plays a key role in the Warburg effect and proliferation of L-02 cells induced by arsenic, and also might be involved in arsenic-induced upregulation of GLUT1 and LDHA. This study provides a theoretical basis for further elucidating the carcinogenic mechanism of arsenic.

Keywords

Arsenic; Cell proliferation; Extracellular signal regulated kinase 1/2; PKM2; Warburg effect.

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