Basic research

Oxidative damage and malignant migration of hepatocellular carcinoma cells LM3 induced by 14 weeks exposure to sodium arsenite

  • Jinli SUN ,
  • Weiwei SONG ,
  • Ming XU ,
  • Jingquan LI
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  • 1.Department of Food Safety and Toxicology, Shanghai Jiao Tong University School of Public Health, Shanghai 200025, China
    2.College of Biological Science and Technology, Baotou Normal College, Inner Mongolia University of Science and Technology, Baotou 014030, China
LI Jingquan, E-mail: jqli@shsmu.edu.cn.

Received date: 2022-09-29

  Accepted date: 2022-11-05

  Online published: 2022-12-28

Supported by

Experimental Technical Team Construction Project of Shanghai Municipal Education Commission(BJ1-3000-22-0090)

Abstract

Objective ·To investigate the effects of different doses of sodium arsenite exposure on human hepatocellular carcinoma cell line LM3, and analyze the underlying potential molecular mechanisms of sodium arsenite-mediated promotion of malignant migration of LM3 cells. Methods ·The model of sodium arsenite exposure was established by continuously culturing LM3 cells in high DMEM glucose medium containing 0, 1 and 10 μmol/L sodium arsenite for 14 weeks. After exposure, the sodium arsenite-containing cell culture medium was discarded, and the cells were allowed to recover in sodium arsenite-free culture medium for one week. Cells from the same batch without sodium arsenite treatment were used as the control group. Subsequently, CCK-8 proliferation assay, Transwell migration assay, and DCFH-DA reactive oxygen species fluorescence probe experiments were performed to verify the effect of chronic sodium arsenite exposure on LM3 cell proliferation, migration, and oxidative stress. Real-time quantitative PCR and Western blotting were performed to elucidate the effect of chronic sodium arsenite exposure on the expression of mRNA and protein related to tumor metastasis in LM3 cells. Results ·Compared with the control group, the sodium arsenite exposure groups (1 μmol/L and 10 μmol/L) had significantly higher reactive oxygen species (ROS) levels (both P>0.05), and sodium arsenite exposure promoted the malignant migration of LM3 cells in a dose-dependent manner (both P>0.05), but sodium arsenite had no significant effect on the proliferation of LM3 cells. Sodium arsenite exposure up-regulated the gene expression levels of vascular endothelial growth factor (VEGF), nicotinamide adenine dinucleotide phosphate oxidases 2 (NOX2), and mitogen-activated protein kinase 8 (MAPK8),and up-regulated the protein expression level of NOX2 and MAPK8. Conclusions ·Sodium arsenite exposure can promote malignant migration and oxidative stress level of LM3 cells, and up-regulate the expression levels of VEGF, NOX2 and MAPK8 genes, suggesting that the mechanism of sodium arsenite promoting malignant migration of LM3 cells may be related to oxidative damage and up-regulation of these genes.

Cite this article

Jinli SUN , Weiwei SONG , Ming XU , Jingquan LI . Oxidative damage and malignant migration of hepatocellular carcinoma cells LM3 induced by 14 weeks exposure to sodium arsenite[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022 , 42(12) : 1677 -1684 . DOI: 10.3969/j.issn.1674-8115.2022.12.004

References

1 SINHA D, PRASAD P. Health effects inflicted by chronic low-level arsenic contamination in groundwater: a global public health challenge[J]. J Appl Toxicol, 2020, 40(1): 87-131.
2 AHMAD S A, KHAN M H, HAQUE M. Arsenic contamination in groundwater in Bangladesh: implications and challenges for healthcare policy[J]. Risk Manag Healthc Policy, 2018, 11: 251-261.
3 GONZáLEZ-HORTA C, BALLINAS-CASARRUBIAS L, SáNCHEZ-RAMíREZ B, et al. A concurrent exposure to arsenic and fluoride from drinking water in Chihuahua, Mexico[J]. Int J Environ Res Public Health, 2015, 12(5): 4587-4601.
4 RODRíGUEZ-LADO L, SUN G F, BERG M, et al. Groundwater arsenic contamination throughout China[J]. Science, 2013, 341(6148): 866-868.
5 ALONSO D L, LATORRE S, CASTILLO E, et al. Environmental occurrence of arsenic in Colombia: a review[J]. Environ Pollut, 2014, 186: 272-281.
6 HUANG L, WU H Y, VAN DER KUIJP T J. The health effects of exposure to arsenic-contaminated drinking water: a review by global geographical distribution[J]. Int J Environ Health Res, 2015, 25(4): 432-452.
7 NELSON G M, AHLBORN G J, ALLEN J W, et al. Transcriptional changes associated with reduced spontaneous liver tumor incidence in mice chronically exposed to high dose arsenic[J]. Toxicology, 2009, 266(1/2/3): 6-15.
8 SMITH A H, MARSHALL G, YUAN Y, et al. Increased mortality from lung cancer and bronchiectasis in young adults after exposure to arsenic in utero and in early childhood[J]. Environ Health Perspect, 2006, 114(8): 1293-1296.
9 CHEN C J, CHEN C W, WU M M, et al. Cancer potential in liver, lung, bladder and kidney due to ingested inorganic arsenic in drinking water[J]. Br J Cancer, 1992, 66(5): 888-892.
10 International Agency for Research on Cancer. IARC monographs on the evaluation of the carcinogenic risk of chemicals to humans[R]. Lyons: World Health Organization, International Agency for Research on Cancer, 1982.
11 LI S, HONG M, TAN H Y, et al. Insights into the role and interdependence of oxidative stress and inflammation in liver diseases[J]. Oxid Med Cell Longev, 2016, 2016: 4234061.
12 WAALKES M P, LIU J, WARD J M, et al. Animal models for arsenic carcinogenesis: inorganic arsenic is a transplacental carcinogen in mice[J]. Toxicol Appl Pharmacol, 2004, 198(3): 377-384.
13 SADAF N, KUMAR N, ALI M, et al. Arsenic trioxide induces apoptosis and inhibits the growth of human liver cancer cells[J]. Life Sci, 2018, 205: 9-17.
14 SANTRA A, MAITI A, CHOWDHURY A, et al. Oxidative stress in liver of mice exposed to arsenic-contaminated water[J]. Indian J Gastroenterol, 2000, 19(3): 112-115.
15 DITZEL E J, LI H, FOY C E, et al. Altered hepatic transport by fetal arsenite exposure in diet-induced fatty liver disease[J]. J Biochem Mol Toxicol, 2016, 30(7): 321-330.
16 DITZEL E J, NGUYEN T, PARKER P, et al. Effects of arsenite exposure during fetal development on energy metabolism and susceptibility to diet-induced fatty liver disease in male mice[J]. Environ Health Perspect, 2016, 124(2): 201-209.
17 CAO M Q, YOU A B, CUI W, et al. Cross talk between oxidative stress and hypoxia via thioredoxin and HIF-2α drives metastasis of hepatocellular carcinoma[J]. FASEB J, 2020, 34(4): 5892-5905.
18 ZHANG C, ZHAO J, ZHAO J, et al. CYP2E1-dependent upregulation of SIRT7 is response to alcohol mediated metastasis in hepatocellular carcinoma[J]. Cancer Gene Ther, 2022: 2022Jul28.
19 戴翔宇, 陈超, 王大朋, 等. 缺氧诱导因子1α在砷所致人肝细胞上皮间质转化及其恶性转化中的作用[J].中华预防医学杂志, 2018, 52(10): 988-993.
19 DAI X Y, CHEN C, WANG D P, et al. Hypoxia-inducible factor-1α is involved in arsenite-induced epithelial-mesenchymal transition and malignant transformation of human liver epithelial cells via regulating Snail[J]. Chin J Prev Med, 2018, 52(10): 988-993.
20 LEE H Y, NGA H T, TIAN J W, et al. Mitochondrial metabolic signatures in hepatocellular carcinoma[J]. Cells, 2021, 10(8): 1901.
21 HSUEH Y M, CHIOU H Y, HUANG Y L, et al. Serum β-carotene level, arsenic methylation capability, and incidence of skin cancer[J]. Cancer Epidemiol Biomarkers Prev, 1997, 6(8): 589-596.
22 NURCHI V M, DJORDJEVIC A B, CRISPONI G, et al. Arsenic toxicity: molecular targets and therapeutic agents[J]. Biomolecules, 2020, 10(2): 235.
23 YU D. A physiologically based pharmacokinetic model of inorganic arsenic[J]. Regul Toxicol Pharmacol, 1999, 29(2 Pt 1): 128-141.
24 PERSON R J, NGALAME N N, MAKIA N L, et al. Chronic inorganic arsenic exposure in vitro induces a cancer cell phenotype in human peripheral lung epithelial cells[J]. Toxicol Appl Pharmacol, 2015, 286(1): 36-43.
25 SUN J, SHI L, XIAO T, et al. microRNA-21, via the HIF-1α/VEGF signaling pathway, is involved in arsenite-induced hepatic fibrosis through aberrant cross-talk of hepatocytes and hepatic stellate cells[J]. Chemosphere, 2021, 266: 129177.
26 PAN X J, DAI Y J, LI X, et al. Inhibition of arsenic-induced rat liver injury by grape seed exact through suppression of NADPH oxidase and TGF-β/Smad activation[J]. Toxicol Appl Pharmacol, 2011, 254(3): 323-331.
27 LEE K M, HWANG M K, LEE D E, et al. Protective effect of quercetin against arsenite-induced COX-2 expression by targeting PI3K in rat liver epithelial cells[J]. J Agric Food Chem, 2010, 58(9): 5815-5820.
28 WANG W J, CHENG S, ZHANG D F. Association of inorganic arsenic exposure with liver cancer mortality: a meta-analysis[J]. Environ Res, 2014, 135: 120-125.
29 SANTRA A, MAITI A, DAS S, et al. Hepatic damage caused by chronic arsenic toxicity in experimental animals[J]. J Toxicol Clin Toxicol, 2000, 38(4): 395-405.
30 SINHA D, PRASAD P. Health effects inflicted by chronic low-level arsenic contamination in groundwater: a global public health challenges[J]. J Appl Toxicol, 2020,40(1): 87-131.
31 HU Y Y, WANG S P, WANG A Q, et al. Antioxidant and hepatoprotective effect of Penthorum chinense Pursh extract against t-BHP-induced liver damage in L02 cells[J]. Molecules, 2015, 20(4): 6443-6453.
32 LI S, TAN H Y, WANG N, et al. The role of oxidative stress and antioxidants in liver diseases[J]. Int J Mol Sci, 2015, 16(11): 26087-26124.
33 MARTNER A, AYDIN E, HELLSTRAND K. NOX2 in autoimmunity, tumor growth and metastasis[J]. J Pathol, 2019, 247(2): 151-154.
34 金子林, 郭锡熔, 周旭华, 等. MAPK8基因在小鼠源性3T3-L1脂肪细胞诱导分化中表达水平的变化[J]. 中国儿童保健杂志, 2004, 12(4): 327-329.
34 JIN Z L, GUO X R, ZHOU X H, et al. Changes of MAPK8 gene expression during mouse 3T3-L1preadipocyte differentiation[J]. Chin J Child Health Care, 2004, 12(4): 327-329.
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