Carcinogenesis, Teratogenesis & Mutagenesis ›› 2020, Vol. 32 ›› Issue (4): 298-303,308.doi: 10.3969/j.issn.1004-616x.2020.04.009

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Silencing of c-fos gene on expression of oncogenes and apoptosis genes in HBE cells exposed to PM2.5

CAI Ying1,2, ZHENG Kai1,2, QIN Shuangjian2,3, LI Boru2,3, YU Shuyuan2, LIU Ning2, JI Jiajia2, ZHANG Zhaohui1, XU Xinyun2   

  1. 1. School of Public Health, University of South China, Hengyang 421001, Hunan;
    2. Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, Guangdong;
    3. Xiangya School of Public Health, Central South University, Changsha 410078, Hunan, China
  • Received:2020-04-21 Revised:2020-07-01 Online:2020-07-31 Published:2020-08-01

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Abstract: OBJECTIVE: To explore the effect of c-fos gene silencing on expression of oncogenes and apoptosis-related genes in human bronchial epithelial (HBE) cells which were exposed to PM2.5. METHODS: According to the c-fos gene mRNA sequence provided by GenBank,interfering sequences were designed and synthesized,and the recombinant lentiviral vector was transfected into HBE cells. Real-time quantitative PCR (qPCR) and Western blot were used to identify the silencing of c-fos gene. HBE cells and c-fos gene-silenced cells were exposed to PM2.5 suspension with a concentration of 50 μg/mL for 24 h,and a non-exposed control group was established. The mRNA levels and protein levels of oncogenes including c-myc,k-ras,c-fos,p53 and apoptosis-related genes including Caspase-3,Caspase-8 were detected by qPCR and Western blot. RESULTS: In HBE cells transfected with c-fos-shRNA lentivirus,qPCR and Western blot analyses revealed that expression of c-fos gene mRNA was 70.1% lower,and expression of c-fos protein was 53.7% lower than that of control group HBE cells. After HBE cells were treated with PM2.5, mRNA expression of c-myc, k-ras, c-fos, Caspase-3,and Caspase-8 were increased by 30.46%,27.17%,and 50.78%,71.91%,74.16%,respectively, compared with the untreated HBE cells,p53 mRNA expression level decreased by 12.27% (P < 0.05). After the c-fos gene silenced cells were treated with PM2.5, mRNA expression of c-myc, k-ras, Caspase-3, and Caspase-8 decreased by 13.08%, 5.39%, 25.03%, 21.37%, respectively, and p53 mRNA decreased by 53.39%,compared with the untreated group. In comparison with normal HBE cells,mRNA expression of c-myc,k-ras,p53,c-fos,Caspase-3,and Caspase-8 were significantly reduced(P < 0.05). Furthermore,protein level of c-myc also showed a decreasing trend after c-fos gene-silenced cells were exposed to PM2.5,compared with that of normal HBE cells. CONCLUSION: c-fos gene-silenced cells were successfully constructed. PM2.5 promoted expression of oncogenes and apoptotic genes in HBE cells. On the other hand,c-fos gene silencing inhibited the expression of oncogenes and apoptotic genes after the PM2.5 treatment in HBE cells.

Key words: c-fos, PM2.5, human bronchial epithelial cells, gene silence, oncogene, apoptosis gene

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