Carcinogenesis, Teratogenesis & Mutagenesis ›› 2022, Vol. 34 ›› Issue (3): 192-199,205.doi: 10.3969/j.issn.1004-616x.2022.03.005

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Effects of four chemicals on the expression of oxidative stress-related genes in HaCaT cells

DUAN Huijuan1,2, SUN Zhaogang1,2, HAO Weidong3,4, WEI Xuetao3,4, CHU Hongqian1,2   

  1. 1. Beijing Key Laboratory of Drug-resistant Tuberculosis Research, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing 101149;
    2. Laboratory of Translational Medicine, Beijing Chest Hospital, Capital Medical University, Beijing 101149;
    3. Department of Toxicology, School of Public Health, Peking University, Beijing 100191;
    4. Beijing Key Laboratory of Food Safety Toxicology Research and Evaluation, Beijing 100191
  • Received:2021-10-12 Revised:2022-02-22 Published:2022-06-10

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Abstract: OBJECTIVE: To investigate effects of hydrogen peroxide,curcumin,quercetin and tert-butyl hydroquinone(tBHQ) on expressions of 84 oxidative stress-related genes. METHODS: HaCaT cells in logarithmic growth phase were used,and different concentrations of hydrogen peroxide,curcumin,quercetin and tert-butyl hydroquinone were applied.The concentrations were:hydrogen peroxide (0.5 and 1.25 mmol/L),curcumin (5 and 20μmol/L),quercetin (200 and 500μmol/L) and tert-butyl hydroquinone (10 and 50μmol/L) for 4 h and 24 h,respectively.Cytotoxicity of the test substances were evaluated by tetramethylthiazole blue (MTT).RNA samples were extracted and their qualities were evaluated.Then,expression levels of 84 oxidative stress-related genes in HaCaT cells were detected by quantitative real-time PCR (qPCR). RESULTS: MTT assay showed that the survival rate of HaCaT cells was less than 80% when the concentrations of hydrogen peroxide,curcumin and tert-butyl hydroquinone were>1 mmol/L,>10μmol/L and>50μmol/L,respectively.Quercetin did not show obvious cytotoxicity at the concentration of 1 000μmol/L.Resultsof qPCR showed that the four chemicals affected expressions of oxidative stress-related genes in HaCaT cells at different concentrations and at different times.The expression levels of some oxidative stress-related genes were increased,and otherswere decreased.Hydrogen peroxide increased expression of HMOX1,SPINK1 (1.25 mmol/L,4 h) but reduced that of CCL5 (1.25 mmol/L,24 h).Curcumin increased HMOX1,ALB (20μmol/L,4 h)expression but reduce that of GSR (20μmol/L,4 h) and other genes.Quercetin increased ALB,HMOX1 (500μmol/L,4 h) expressions but reduced expression of SEPP1 (500μmol/L,4 h).tert-butyl hydroquinone increased expression of EPX,HMOX1 (10μmol/L,4 h) but reduced that of AOX1 (10μmol/L,24 h),etc. CONCLUSION: Exposure to hydrogen peroxide,curcumin,quercetin and tert-butyl hydroquinone caused changes in expression of oxidative stress-related genes,especially increased expression the HMOX1 gene.Moreover,higher concentrations of the chemicals caused more changes than lower doses.

Key words: oxidative stress, hydrogen peroxide, curcumin, quercetin, tert-butyl hydroquinone

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