| Objective:Environmental arsenic exposure is one of the major public health problems in the world,and the mechanism of arsenic carcinogenesis is still unclear.Arsenic can cause damage to a variety of systemic organs,and lung is the main target organ for arsenic carcinogenesis.Epidemiology and in vivo and in vitro studies have proved that arsenic is closely related to lung cancer.An important marker of metabolic reprogramming of tumors,is a series of changes in metabolic characteristics mainly due to Warburg effect due to the structural and functional changes of some genes in tumor cells,which are mainly manifested as enhanced glycolysis,increased glucose uptake and consumption,enhanced synthesis of lipids and protein,as well as increased uptake and catabolism of amino acids such as glutamine.These metabolic changes provide adequate energy supply for abnormal proliferation of tumor cells.By elucidating the alteration of metabolic spectra in malignant transformation of cell induced by chronic inorganic arsenic,we can gain a significant foundation for examining the carcinogenic mechanism of arsenic.The most common metabolic change in tumor cells is enhanced glycolysis,which is mainly caused by the enhanced expression or activity of key glycolytic enzymes.HK2 the key rate-limiting enzyme in glycolytic pathways,is hexokinase.This study employed a BEAS-2B cell malignant transformation model exposed to chronic arsenic to conduct targeted metabolome detection,beginning with the glycolytic metabolic pathway,in order to investigate effect of glycolytic pathway on malignant transformation cells induced by chronic inorganic arsenic and its mechanism and to offer fresh insights and is used for diagnosing,preventing and treating tumors induced by chronic inorganic arsenic.Methods:1.Establishment of human bronchial epithelial cell malignant transformation model induced by chronic inorganic arsenic:BEAS-2B cells were incubated with 0.1μM Na As O2for 44 weeks and the normal medium was used in the control group.Testing of cell proliferation,migration,and invasion ability was conducted through CCK8,scratch healing test,and transwell.2.Cellular metabolome detection:The ultra-performance liquid chromatographic system is used for targeted detection of total differential metabolites in cells.3.RT-qPCR and Western blotting were employed to identify metabolic enzymes associated with glycolytic pathways in malignant transformed BEAS-2B cells after 44 weeks of persistent arsenic exposure,thereby screening for key glycolytic enzymes.4.The malignant phenotype caused by chronic arsenic exposure was revealed in BEAS-2B cells after 44 weeks of exposure,when they were treated with a culture solution without glucose or glycolysis inhibitor2-DG.CCK8,scratch healing experiment and transwell to detect cell proliferation,migration and invasion.Results:1.Chronic inorganic arsenic exposure to BEAS-2B cells caused morphological changes of spindle-like and diamond-like nature,with a marked enhancement in cell proliferation,migration,and invasion capabilities（P<0.05）.2.The targeted metabolome method was employed to analyze more than 600 common related metabolites,including carbohydrates,organic acids,amino acids,bile acids,indoles,purine nucleotides,and lipids,through the utilization of an ultra-performance liquid chromatography system for the detection of cell metabolome.3.The pathway of significantly different metabolites was analyzed in KEGG,and most metabolites in the glycolysis pathway were significantly changed.4.The ability to proliferate,migrate,and invade inorganic arsenic-intoxicating cells was significantly reduced in the glucose-deprived or 2-DG-treated groups when compared to normally cultured inorganic arsenic-intoxicating cells.5.The protein and mRNA expression levels of enzymes related to glycolytic metabolic pathway in BEAS-2B cells with chronic inorganic arsenic was observed,which led to malignant transformation of cells.Among them,the up-regulation of HK2 was obvious（P<0.05）.Conclusion:The expression of HK2,glucose dependence on growth,and glycolytic activity of human bronchial epithelial cells when exposed to inorganic arsenic are all indicative of malignant transformation,which may be the cause of the cells’malignant transformation. |
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