DOI QR코드

DOI QR Code

The Inhibitory Effect of NLRP3 Deficiency in Hepatocellular Carcinoma SK-Hep1 Cells

  • Choi, Wonhyeok (College of Pharmacy, Duksung Women's University) ;
  • Cho, Hyosun (College of Pharmacy, Duksung Women's University)
  • 투고 : 2021.07.15
  • 심사 : 2021.10.12
  • 발행 : 2021.12.28

초록

The NLRP3 (nucleotide-binding domain, leucine-rich repeat family pyrin domain containing 3) inflammasome plays an important role in the initiation of inflammatory responses, through the recognition of pathogen-associated molecular patterns and tumor progression, including tumor growth and metastasis. In this study, we examined the effects of defective NLRP3 on the growth, migration, and invasiveness of hepatocellular carcinoma (HCC) SK-Hep1 cell. First, HCC SK-Hep1 cells were transfected with human NLRP3 targeting LentiCRISPRv2 vector using the CRISPR-Cas9 system, and NLRP3 deficiency was confirmed by RT-qPCR and western blotting. NLRP3 deficient SK-Hep1 cells showed delayed cell growth and decreased protein expression of PI3K, p-AKT, and pNF-κB when compared to NLRP3 complete SK-Hep1 cells. In addition, NLRP3 deficiency arrested the cell cycle at G1 phase through an increase in p21 and a reduction in CDK6. NLRP3 deficient SK-Hep1 cells also showed significantly delayed cell migration, invasion, and wound healing. The expression of epithelial-mesenchymal transition signaling molecules, such as N-cadherin and MMP-9, was found to be dramatically decreased in NLRP3 deficient SK-Hep1 cells compared to NLRP3 complete SK-Hep1 cells.

키워드

과제정보

This research was supported by Duksung Women's University Research Grants 2020-3000005168.

참고문헌

  1. Balogh J, Victor III D, Asham EH, Burroughs SG, Boktour M, Saharia A, et al. 2016. Hepatocellular carcinoma: a review. J. Hepatocell. Carcinoma 3: 41-53. https://doi.org/10.2147/JHC.S61146
  2. Lee HE, Lee JY, Yang G, Kang HC, Cho YY, Lee HS, et al. 2019. Inhibition of NLRP3 inflammasome in tumor microenvironment leads to suppression of metastatic potential of cancer cells. Sci. Rep. 9: 1-9. https://doi.org/10.1038/s41598-018-37186-2
  3. 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. https://doi.org/10.3322/caac.21492
  4. Eun JR, Jung YJ, Zhang Y, Zhang YH, Tschudyseney B, Ramsamooj R, et al. 2014. Hepatoma SK Hep-1 cells exhibit characteristics of oncogenic mesenchymal stem cells with highly metastatic capacity. PLoS One 9: 110744.
  5. Latz E. 2010. The inflammasomes: mechanisms of activation and function. Curr. Opin. Immunol. 22: 28-33. https://doi.org/10.1016/j.coi.2009.12.004
  6. Moossavi M, Parsamanesh N, Bahrami A, Atkin SL, Sahebkar A. 2018. Role of the NLRP3 inflammasome in cancer. Mol. Cancer 17: 158. https://doi.org/10.1186/s12943-018-0900-3
  7. Shaima'a Hamarsheh RZ. 2020. NLRP3 inflammasome activation in cancer: a double-edged sword. Front. Immunol. 11: 1444. https://doi.org/10.3389/fimmu.2020.01444
  8. Wei Q, Mu K, Li T, Zhang Y, Yang Z, Jia X, et al. 2014. Deregulation of the NLRP3 inflammasome in hepatic parenchymal cells during liver cancer progression. Lab. Invest. 94: 52-62. https://doi.org/10.1038/labinvest.2013.126
  9. Wree A, Eguchi A, McGeough MD, Pena CA, Johnson CD, Canbay A, et al. 2014. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice. Hepatology 59: 898-910. https://doi.org/10.1002/hep.26592
  10. Ballestar E, Esteller M. 2008. Epigenetic gene regulation in cancer. Adv. Genet. 61: 247-267. https://doi.org/10.1016/S0065-2660(07)00009-0
  11. Hui AM, Makuuchi M, Li X. 1998. Cell cycle regulators and human hepatocarcinogenesis. Hepatogastroenterology 45: 1635-1642.
  12. Diepenbruck M, Christofori G. 2016. Epithelial-mesenchymal transition (EMT) and metastasis: yes, no, maybe?. Curr. Opin. Cell Biol. 43: 7-13. https://doi.org/10.1016/j.ceb.2016.06.002
  13. Wan S, Meyer AS, Weiler SME, Rupp C, Toth M, Sticht C, et al. 2018. Cytoplasmic localization of the cell polarity factor scribble supports liver tumor formation and tumor cell invasiveness. Hepatology 67: 1842-1856. https://doi.org/10.1002/hep.29669
  14. Thiery JP, Acloque H, Huang RY, Nieto MA. 2009. Epithelial-mesenchymal transitions in development and disease. Cell 139: 871-890. https://doi.org/10.1016/j.cell.2009.11.007
  15. Zeisberg M, Neilson EG. 2009. Biomarkers for epithelial-mesenchymal transitions. J. Clin. Invest. 119: 1429-1437. https://doi.org/10.1172/JCI36183
  16. Kitamura T, Taketo MM. 2007. Keeping out the bad guys: gateway to cellular target therapy. Cancer Res. 67: 10099-10102. https://doi.org/10.1158/0008-5472.CAN-07-2100
  17. Kalluri R, Weinberg RA. 2009. The basics of epithelial-mesenchymal transition. J. Clin. Invest. 119: 1420-1428. https://doi.org/10.1172/JCI39104
  18. Karki R, Man SM, Kanneganti TD. 2017. Inflammasomes and cancer. Cancer Immunol. Res. 5: 94-99. https://doi.org/10.1158/2326-6066.CIR-16-0269
  19. Imaeda AB, Watanabe A, Sohail MA, Mahmood S, Mohamadnejad M, Sutterwala FS, et al. 2009. Acetaminophen-induced hepatotoxicity in mice is dependent on Tlr9 and the Nalp3 inflammasome. J. Clin. Invest. 119: 305-314. https://doi.org/10.1172/JCI35958
  20. Fan SH, Wang YY, Lu J, Zheng YL, Wu DM, Li MQ, et al. 2014. Luteoloside suppresses proliferation and metastasis of hepatocellular carcinoma cells by inhibition of NLRP3 inflammasome. PLoS One 9: e89961. https://doi.org/10.1371/journal.pone.0089961
  21. Wan L, Yuan X, Liu M, Xue B. 2018. miRNA-223-3p regulates NLRP3 to promote apoptosis and inhibit proliferation of hep3B cells. Exp. Ther. Med. 15: 2429-2435.
  22. Zhang Y, Yang H, Sun M, He T, Liu Y, Yang X, et al. 2020. Alpinumisoflavone suppresses hepatocellular carcinoma cell growth and metastasis via NLRP3 inflammasome-mediated pyroptosis. Pharmacol. Rep. 72: 1370-1382. https://doi.org/10.1007/s43440-020-00064-8
  23. Wang H, Luo Q, Feng X, Zhang R, Li J, Chen F. 2018. NLRP3 promotes tumor growth and metastasis in human oral squamous cell carcinoma. BMC Cancer 18: 500. https://doi.org/10.1186/s12885-018-4403-9
  24. Zhang L, Li H, Zang Y, Wang F. 2019. NLRP3 inflammasome inactivation driven by miR-223-3p reduces tumor growth and increases anticancer immunity in breast cancer. Mol. Med. Rep. 19: 2180-2188.
  25. Tian X, Zhang S, Zhang Q, Kang L, Ma C, Feng L, et al. 2020. Resveratrol inhibits tumor progression by down-regulation of NLRP3 in renal cell carcinoma. J. Nutr. Biochem. 85: 108489. https://doi.org/10.1016/j.jnutbio.2020.108489
  26. Xue L, Lu B, Gao B, Shi Y, Xu J, Yang R, et al. 2019. NLRP3 promotes glioma cell proliferation and invasion via the interleukin-1β/ NF-κB p65 signals. Oncol. Res. 27: 557-564. https://doi.org/10.3727/096504018X15264647024196
  27. Zhou T, Xiang DK, Li SN, Yang LH, Gao LF, Feng C. 2018. MicroRNA-495 ameliorates cardiac microvascular endothelial cell injury and inflammatory reaction by suppressing the NLRP3 inflammasome signaling pathway. Cell. Physiol. Biochem. 49: 798-815. https://doi.org/10.1159/000493042
  28. Weichand B, Popp R, Dziumbla S, Mora J, Strack E, Elwakeel E, et al. 2017. S1PR1 on tumor-associated macrophages promotes lymphangiogenesis and metastasis via NLRP3/IL-1β. J. Exp. Med. 214: 2695-2713. https://doi.org/10.1084/jem.20160392
  29. Deng Q, Geng Y, Zhao L, Li R, Zhang Z, Li K, et al. 2019. NLRP3 inflammasomes in macrophages drive colorectal cancer metastasis to the liver. Cancer Lett. 442: 21-30. https://doi.org/10.1016/j.canlet.2018.10.030
  30. Chen YC, Shen SC, Lee WR, Lin HY, Ko CH, Shih CM, et al. 2002. Wogonin and fisetin induction of apoptosis through activation of caspase 3 cascade and alternative expression of p21 protein in hepatocellular carcinoma cells SK-HEP-1. Arch. Toxicol. 76: 351-359. https://doi.org/10.1007/s00204-002-0346-6
  31. Hanahan D, Weinberg RA. 2011. Hallmarks of cancer: the next generation. Cell 144: 646-674. https://doi.org/10.1016/j.cell.2011.02.013
  32. Xu G, Zhu L, Wang Y, Shi Y, Gong A, Wu C. 2017. Stattic enhances radiosensitivity and reduces radio-induced migration and invasion in HCC cell lines through an apoptosis pathway. Biomed Res. Int. 2017: 1832494.
  33. Saitoh M. 2018. Involvement of partial EMT in cancer progression. J. Biochem. 164: 257-264. https://doi.org/10.1093/jb/mvy047
  34. Shao X, Lei Z, Zhou C. 2020. NLRP3 promotes colorectal cancer cell proliferation and metastasis via regulating epithelial mesenchymal transformation. Anticancer Agents Med. Chem. 20: 820-827. https://doi.org/10.2174/1871520620666200220112741