Carcinogenesis, Teratogenesis & Mutagenesis ›› 2021, Vol. 33 ›› Issue (2): 129-135,142.doi: 10.3969/j.issn.1004-616x.2021.02.008

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Effects of p38MAPK gene on PM2.5-induced expression of oncogenes and apoptosis genes in HK-2 cells

LI Boru1,2, QIN Shuangjian1,2, LI Runbing2,3, CAI Ying2,3, ZHENG Kai2,3, ZENG Ming1, XIAO Fang1, XU Xinyun2   

  1. 1. Xiangya School of Public Health, Central South University, Changsha 410078, Hunan;
    2. Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, Guangdong;
    3. School of Public Health, University of South China, Hengyang 421001, Hunan, China
  • Received:2020-08-21 Revised:2020-11-10 Online:2021-03-30 Published:2021-04-12

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Abstract: OBJECTIVE: To investigate effects of p38MAPK gene on expression of oncogenes and apoptosis related genes in human kidney epithelial cells (HK-2 cells) which had been exposed to PM2.5. METHODS: According to the mRNA sequence of p38MAPK gene provided by GenBank,interference sequences were designed and synthesized,and a recombinant lentiviral vector was constructed and was transfected into HK-2 cells to construct p38MAPK gene-silenced cells. Real-time fluorescent quantitative PCR (qPCR) and western blot were used to identifyeffects from p38MAPK gene silencing. HK-2 cells and p38MAPK gene-silenced cells were treated with 50 μg/mL PM2.5 suspension for 24 h,and qPCR and western blot were used to detect their expression of oncogenes (c-myc,c-fos and p53) and apoptosis-related genes (Caspase-8,Caspase-9 and Bcl-2). RESULTS: p38MAPK mRNA expression levels in p38MAPK-silenced cells were inhibited by 58.50% and p38MAPK protein expression levels by 51.33% when compared with the normal HK-2 cells (P<0.01). These results indicate that the p38MAPK silenced cells were successfully constructed. The qPCR results showed that when compared with the HK-2 cells in the control group,the PM2.5-exposed cells indicated the mRNA of c-myc,c-fos,Caspase-8 and Casepase-9 in expressions were significantly increased by 39.89%,15.12%,19.47% and 15.45%,respectively,and p53 and Bcl-2 mRNA expressions were significantly decreased by 21.54% and 31.77% respectively (P<0.05). When compared with the PM2.5 exposed HK-2 cells,the PM2.5-exposed p38MAPK-silenced cells showed that the mRNA of c-myc,c-fos,p53,Caspase-8 and Caspase-9 expressions were significantly decreased by 84.55%、63.55%、34.49%、37.19% and 54.97% respectively,(P<0.05). At the protein level,when compared with the HK-2 cells in the control group,the PM2.5 exposed cells showed that the protein levels of c-myc,and Caspase-8 were increased and that of Bcl-2 were decreased. When compared with the PM2.5-exposed HK-2 cells,the PM2.5-exposed p38MAPK-silenced cells showed that the mRNA levels of c-myc,Caspase-8 and Bcl-2 were decreased. CONCLUSION: p38MAPK-silenced cells were successfully constructed in this study. Our data show that PM2.5 promoted the expression of oncogenes and apoptotic-related genes in HK-2 cells,and the p38MAPK gene was involved in the cytotoxicity of PM2.5.

Key words: p38MAPK, PM2.5, human kidney epithelial cells, gene silence, oncogenes, apoptosis genes

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