Involvement of p38 MAPK pathway in benzo(a)pyrene-induced human hepatoma cell migration and invasion

Abstract

The objective of this study was to investigate the potential role of p38 mitogen-activated protein kinases (MAPK) in benzo(a)pyrene (BaP)-induced hepatoma cell migration and invasion. Western blot assay was applied to detect the expression of proteins. qRT-PCR assay was used to measure the expression of mRNA. Wound healing assay and Transwell invasion assay were performed to evaluate cell migratory ability and cell invasive ability, respectively. Our data showed that BaP exposure increased the expression of p-p38 protein in human hepatoma HepG2 cells. Exposure to BaP facilitated HepG2 cell migration and invasion, which could be blocked by p38 MAPK inhibitors. In addition, BaP exposure induced upregulation of MMP9 mRNA expression, which was modulated by p-p38. In conclusion, p38 MAPK pathway was involved in BaP-induced hepatoma cell migration and invasion.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. Arnould C, Lelievre-Pegorier M, Ronco P, Lelongt B (2009) MMP9 limits apoptosis and stimulates branching morphogenesis during kidney development. J Am Soc Nephrol 20:2171–2180

    CAS  Google Scholar 

  2. Ba Q, Li J, Huang C, Qiu H, Chu R, Zhang W, Xie D, Wu Y, Wang H (2015) Effects of benzo[a]pyrene exposure on human hepatocellular carcinoma cell angiogenesis, metastasis, and NF-kappaB signaling. Environ Health Perspect 123:246–254

    CAS  Google Scholar 

  3. Begeman CR, Colucci JM (1968) Benzo(a)pyrene in gasoline partially persists in automobile exhaust. Science 161:271

    CAS  Google Scholar 

  4. Bertuccio P, Turati F, Carioli G, Rodriguez T, La Vecchia C, Malvezzi M, Negri E (2017) Global trends and predictions in hepatocellular carcinoma mortality. J Hepatol 67:302–309

    Google Scholar 

  5. Bolotina NA, Gasparian AV, Dubovaja TK, Evteev VA, Kobliakov VA (2007) Benzo[a]pyrene-dependent activation of transcription factors NF-kappaB and AP-1 related to tumor promotion in hepatoma cell cultures. Biochemistry (Mosc) 72:552–557

    CAS  Google Scholar 

  6. Bonney EA (2017): Mapping out p38MAPK. Am J Reprod Immunol 77

    Google Scholar 

  7. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68:394–424

    Google Scholar 

  8. Chen X, Bo L, Zhao X, Chen Q (2015) MicroRNA-133a inhibits cell proliferation, colony formation ability, migration and invasion by targeting matrix metallopeptidase 9 in hepatocellular carcinoma. Mol Med Rep 11:3900–3907

    CAS  Google Scholar 

  9. Cheng G, Gao F, Sun X, Bi H, Zhu Y (2016) Paris saponin VII suppresses osteosarcoma cell migration and invasion by inhibiting MMP2/9 production via the p38 MAPK signaling pathway. Mol Med Rep 14:3199–3205

    CAS  Google Scholar 

  10. Coulthard LR, White DE, Jones DL, McDermott MF, Burchill SA (2009) p38(MAPK): stress responses from molecular mechanisms to therapeutics. Trends Mol Med 15:369–379

    CAS  Google Scholar 

  11. Cuadrado A, Nebreda AR (2010) Mechanisms and functions of p38 MAPK signalling. Biochem J 429:403–417

    CAS  Google Scholar 

  12. Cui H, Gu X, Chen J, Xie Y, Ke S, Wu J, Golovko A, Morpurgo B, Yan C, Phillips TD, Xie W, Luo J, Zhou Z, Tian Y (2017) Pregnane X receptor regulates the AhR/Cyp1A1 pathway and protects liver cells from benzo-[a]-pyrene-induced DNA damage. Toxicol Lett 275:67–76

    CAS  Google Scholar 

  13. del Barco Barrantes I, Nebreda AR (2012) Roles of p38 MAPKs in invasion and metastasis. Biochem Soc Trans 40:79–84

    Google Scholar 

  14. Feng M, Wang Y, Chen K, Bian Z, Jinfang W, Gao Q (2014) IL-17A promotes the migration and invasiveness of cervical cancer cells by coordinately activating MMPs expression via the p38/NF-kappaB signal pathway. PloS one 9:e108502

    Google Scholar 

  15. Gelboin HV (1980) Benzo[alpha]pyrene metabolism, activation and carcinogenesis: role and regulation of mixed-function oxidases and related enzymes. Physiol Rev 60:1107–1166

    CAS  Google Scholar 

  16. Guo Y, Jiang M, Zhao X, Gu M, Wang Z, Xu S, Yue W (2018) Cyclophilin A promotes non-small cell lung cancer metastasis via p38 MAPK. Thorac Cancer 9:120–128

    CAS  Google Scholar 

  17. Hsieh YH, Wu TT, Huang CY, Hsieh YS, Hwang JM, Liu JY (2007) p38 mitogen-activated protein kinase pathway is involved in protein kinase Calpha-regulated invasion in human hepatocellular carcinoma cells. Cancer Res 67:4320–4327

    CAS  Google Scholar 

  18. Hsieh MJ, Chen KS, Chiou HL, Hsieh YS (2010) Carbonic anhydrase XII promotes invasion and migration ability of MDA-MB-231 breast cancer cells through the p38 MAPK signaling pathway. Eur J Cell Biol 89:598–606

    CAS  Google Scholar 

  19. Hsu HH, Liu CJ, Shen CY, Chen YJ, Chen LM, Kuo WH, Lin YM, Chen RJ, Tsai CH, Tsai FJ, Huang CY (2012) p38alpha MAPK mediates 17beta-estradiol inhibition of MMP-2 and -9 expression and cell migration in human lovo colon cancer cells. J Cell Physiol 227:3648–3660

    CAS  Google Scholar 

  20. Huang Q, Lan F, Wang X, Yu Y, Ouyang X, Zheng F, Han J, Lin Y, Xie Y, Xie F, Liu W, Yang X, Wang H, Dong L, Wang L, Tan J (2014) IL-1beta-induced activation of p38 promotes metastasis in gastric adenocarcinoma via upregulation of AP-1/c-fos, MMP2 and MMP9. Mol Cancer 13:18

    Google Scholar 

  21. Huberman E (1978) Mutagenesis and cell transformation of mammalian cells in culture by chemical carcinogens. J Environ Pathol Toxicol 2:29–42

    CAS  Google Scholar 

  22. IARC (2010) Some non-heterocyclic polycyclic aromatic hydrocarbons and some related exposures. IARC Monogr Eval Carcinog Risks Hum 92:1–853

    Google Scholar 

  23. Jacob A, Prekeris R (2015) The regulation of MMP targeting to invadopodia during cancer metastasis. Front Cell Dev Biol 3:4

    Google Scholar 

  24. Kumar B, Koul S, Petersen J, Khandrika L, Hwa JS, Meacham RB, Wilson S, Koul HK (2010) p38 mitogen-activated protein kinase-driven MAPKAPK2 regulates invasion of bladder cancer by modulation of MMP-2 and MMP-9 activity. Cancer Res 70:832–841

    CAS  Google Scholar 

  25. Marrone AK, Tryndyak V, Beland FA, Pogribny IP (2016) MicroRNA responses to the genotoxic carcinogens aflatoxin B1 and benzo[a]pyrene in human HepaRG cells. Toxicol Sci 149:496–502

    CAS  Google Scholar 

  26. Mastral AM, Callen MS, Garcia T, Lopez JM (2001) Benzo(a)pyrene, benzo(a)anthracene, and dibenzo(a,h)anthracene emissions from coal and waste tire energy generation at atmospheric fluidized bed combustion (AFBC). Environ Sci Technol 35:2645–2649

    CAS  Google Scholar 

  27. Min J, Feng Q, Liao W, Liang Y, Gong C, Li E, He W, Yuan R, Wu L (2018) IFITM3 promotes hepatocellular carcinoma invasion and metastasis by regulating MMP9 through p38/MAPK signaling. FEBS Open Bio 8:1299–1311

    CAS  Google Scholar 

  28. Padwal M, Siddique I, Wu L, Tang K, Boivin F, Liu L, Robertson J, Bridgewater D, West-Mays J, Gangji A, Brimble KS, Margetts PJ (2017) Matrix metalloproteinase 9 is associated with peritoneal membrane solute transport and induces angiogenesis through beta-catenin signaling. Nephrol Dial Transplant 32:50–61

    CAS  Google Scholar 

  29. Plaks V, Rinkenberger J, Dai J, Flannery M, Sund M, Kanasaki K, Ni W, Kalluri R, Werb Z (2013) Matrix metalloproteinase-9 deficiency phenocopies features of preeclampsia and intrauterine growth restriction. Proc Natl Acad Sci U S A 110:11109–11114

    CAS  Google Scholar 

  30. Pujada A, Walter L, Patel A, Bui TA, Zhang Z, Zhang Y, Denning TL, Garg P (2017) Matrix metalloproteinase MMP9 maintains epithelial barrier function and preserves mucosal lining in colitis associated cancer. Oncotarget 8:94650–94665

    Google Scholar 

  31. Reed L, Arlt VM, Phillips DH (2018) The role of cytochrome P450 enzymes in carcinogen activation and detoxication: an in vivo-in vitro paradox. Carcinogenesis 39:851–859

    CAS  Google Scholar 

  32. Rubin H (2001) Synergistic mechanisms in carcinogenesis by polycyclic aromatic hydrocarbons and by tobacco smoke: a bio-historical perspective with updates. Carcinogenesis 22:1903–1930

    CAS  Google Scholar 

  33. Shen KH, Hung SH, Yin LT, Huang CS, Chao CH, Liu CL, Shih YW (2010) Acacetin, a flavonoid, inhibits the invasion and migration of human prostate cancer DU145 cells via inactivation of the p38 MAPK signaling pathway. Mol Cell Biochem 333:279–291

    CAS  Google Scholar 

  34. Smith CJ, Perfetti TA, Rumple MA, Rodgman A, Doolittle DJ (2000) “IARC group 2A Carcinogens” reported in cigarette mainstream smoke. Food Chem Toxicol 38:371–383

    CAS  Google Scholar 

  35. Souza T, Jennen D, van Delft J, van Herwijnen M, Kyrtoupolos S, Kleinjans J (2016) New insights into BaP-induced toxicity: role of major metabolites in transcriptomics and contribution to hepatocarcinogenesis. Arch Toxicol 90:1449–1458

    CAS  Google Scholar 

  36. Su Y, Zhao B, Guo F, Bin Z, Yang Y, Liu S, Han Y, Niu J, Ke X, Wang N, Geng X, Jin C, Dai Y, Lin Y (2014) Interaction of benzo[a]pyrene with other risk factors in hepatocellular carcinoma: a case-control study in Xiamen, China. Ann Epidemiol 24:98–103

    Google Scholar 

  37. Sui X, Kong N, Ye L, Han W, Zhou J, Zhang Q, He C, Pan H (2014) p38 and JNK MAPK pathways control the balance of apoptosis and autophagy in response to chemotherapeutic agents. Cancer Lett 344:174–179

    CAS  Google Scholar 

  38. Sun SJ, Wang N, Sun ZW, Chen J, Cui HW (2018) MiR-5692a promotes the invasion and metastasis of hepatocellular carcinoma via MMP9. Eur Rev Med Pharmacol Sci 22:4869–4878

    Google Scholar 

  39. Tormos AM, Talens-Visconti R, Nebreda AR, Sastre J (2013) p38 MAPK: a dual role in hepatocyte proliferation through reactive oxygen species. Free Radic Res 47:905–916

    CAS  Google Scholar 

  40. Umannova L, Machala M, Topinka J, Schmuczerova J, Krcmar P, Neca J, Sujanova K, Kozubik A, Vondracek J (2011) Benzo[a]pyrene and tumor necrosis factor-alpha coordinately increase genotoxic damage and the production of proinflammatory mediators in alveolar epithelial type II cells. Toxicol Lett 206:121–129

    CAS  Google Scholar 

  41. van Kempen LC, Coussens LM (2002) MMP9 potentiates pulmonary metastasis formation. Cancer Cell 2:251–252

    Google Scholar 

  42. van Kesteren PC, Zwart PE, Pennings JL, Gottschalk WH, Kleinjans JC, van Delft JH, van Steeg H, Luijten M (2011) Deregulation of cancer-related pathways in primary hepatocytes derived from DNA repair-deficient Xpa-/-p53+/- mice upon exposure to benzo[a]pyrene. Toxicol Sci 123:123–132

    Google Scholar 

  43. van Kesteren PC, Zwart PE, Schaap MM, Pronk TE, van Herwijnen MH, Kleinjans JC, Bokkers BG, Godschalk RW, Zeilmaker MJ, van Steeg H, Luijten M (2013) Benzo[a]pyrene-induced transcriptomic responses in primary hepatocytes and in vivo liver: toxicokinetics is essential for in vivo-in vitro comparisons. Arch Toxicol 87:505–515

    Google Scholar 

  44. Vesselinovitch SD, Kyriazis AP, Mihailovich N, Rao KV (1975) Conditions modifying development of tumors in mice at various sites by benzo(a)pyrene. Cancer Res 35:2948–2953

    CAS  Google Scholar 

  45. Wang Q, Yu W, Huang T, Zhu Y, Huang C (2016) RUNX2 promotes hepatocellular carcinoma cell migration and invasion by upregulating MMP9 expression. Oncol Rep 36:2777–2784

    CAS  Google Scholar 

  46. Wang Y, Pan T, Li L, Wang H, Zhang D, Yang H (2018a) Benzo(a)pyrene promotes Hep-G2 cell migration and invasion by upregulating phosphorylated extracellular signal-regulated kinase expression. Oncol Lett 15:8325–8332

    Google Scholar 

  47. Wang Y, Xu Y, Yan W, Han P, Liu J, Gong J, Li D, Ding X, Wang H, Lin Z, Tian D, Liao J (2018b) CFIm25 inhibits hepatocellular carcinoma metastasis by suppressing the p38 and JNK/c-Jun signaling pathways. Oncotarget 9:11783–11793

    Google Scholar 

  48. Wang Y, Shi L, Li J, Li L, Wang H, Yang H (2019) Long-term cadmium exposure promoted breast cancer cell migration and invasion by up-regulating TGIF. Ecotoxicol Environ Saf 175:110–117

    CAS  Google Scholar 

  49. Wei W, Li XF, Li XN, Chen XM, Liu AL, Lu WQ (2012) Oxidative stress and cell-cycle change induced by coexposed PCB126 and benzo(a) pyrene to human hepatoma (HepG2) cells. Environ Toxicol 27:316–320

    CAS  Google Scholar 

  50. Wester PW, Muller JJ, Slob W, Mohn GR, Dortant PM, Kroese ED (2012) Carcinogenic activity of benzo[a]pyrene in a 2 year oral study in Wistar rats. Food Chem Toxicol 50:927–935

    CAS  Google Scholar 

  51. Yang F, Yin Y, Wang F, Wang Y, Zhang L, Tang Y, Sun S (2010) miR-17-5p promotes migration of human hepatocellular carcinoma cells through the p38 mitogen-activated protein kinase-heat shock protein 27 pathway. Hepatology 51:1614–1623

    CAS  Google Scholar 

  52. Yang H, Zhang H, Pan T, Wang H, Wang Y (2018) Benzo(a)pyrene promotes migration, invasion and metastasis of lung adenocarcinoma cells by upregulating TGIF. Toxicol Lett 294:11–19

    CAS  Google Scholar 

  53. Yu Y, Li H, Xue B, Jiang X, Huang K, Ge J, Zhang H, Chen B (2014) SDF-1/CXCR7 axis enhances ovarian cancer cell invasion by MMP-9 expression through p38 MAPK pathway. DNA Cell Biol 33:543–549

    CAS  Google Scholar 

  54. Zhu B, Shi S, Ma YG, Fan F, Yao ZZ (2012) Lysophosphatidic acid enhances human hepatocellular carcinoma cell migration, invasion and adhesion through P38 MAPK pathway. Hepatogastroenterology 59:785–789

    CAS  Google Scholar 

Download references

Availability of data and materials

The datasets generated/analyzed during the present study are included in the published article.

Funding

This work was funded by the National Natural Science Foundation of China (No. U1404815) and Henan Province University Science and Technology Innovation Talent Projects (17HASTIT045). The funder had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Author information

Affiliations

Authors

Corresponding authors

Correspondence to Yadong Wang or Haiyan Yang.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Responsible editor: Philippe Garrigues

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Wang, Y., Shi, L., Li, J. et al. Involvement of p38 MAPK pathway in benzo(a)pyrene-induced human hepatoma cell migration and invasion. Environ Sci Pollut Res 26, 35838–35845 (2019). https://doi.org/10.1007/s11356-019-06733-3

Download citation

Keywords

  • Benzo(a)pyrene
  • Migration
  • Invasion
  • p38 MAPK
  • MMP9
  • HepG2 cells