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http://dx.doi.org/10.48022/mbl.2107.07004

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)
Publication Information
Microbiology and Biotechnology Letters / v.49, no.4, 2021 , pp. 594-602 More about this Journal
Abstract
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.
Keywords
NLRP3; SK-Hep1; cell cycle; EMT;
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1 Thiery JP, Acloque H, Huang RY, Nieto MA. 2009. Epithelial-mesenchymal transitions in development and disease. Cell 139: 871-890.   DOI
2 Zeisberg M, Neilson EG. 2009. Biomarkers for epithelial-mesenchymal transitions. J. Clin. Invest. 119: 1429-1437.   DOI
3 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.   DOI
4 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.   DOI
5 Kitamura T, Taketo MM. 2007. Keeping out the bad guys: gateway to cellular target therapy. Cancer Res. 67: 10099-10102.   DOI
6 Kalluri R, Weinberg RA. 2009. The basics of epithelial-mesenchymal transition. J. Clin. Invest. 119: 1420-1428.   DOI
7 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.
8 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.   DOI
9 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.   DOI
10 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.   DOI
11 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.   DOI
12 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.   DOI
13 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.   DOI
14 Hanahan D, Weinberg RA. 2011. Hallmarks of cancer: the next generation. Cell 144: 646-674.   DOI
15 Saitoh M. 2018. Involvement of partial EMT in cancer progression. J. Biochem. 164: 257-264.   DOI
16 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.   DOI
17 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.
18 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.   DOI
19 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.   DOI
20 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.
21 Karki R, Man SM, Kanneganti TD. 2017. Inflammasomes and cancer. Cancer Immunol. Res. 5: 94-99.   DOI
22 Shaima'a Hamarsheh RZ. 2020. NLRP3 inflammasome activation in cancer: a double-edged sword. Front. Immunol. 11: 1444.   DOI
23 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.   DOI
24 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.   DOI
25 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.
26 Latz E. 2010. The inflammasomes: mechanisms of activation and function. Curr. Opin. Immunol. 22: 28-33.   DOI
27 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.   DOI
28 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.   DOI
29 Moossavi M, Parsamanesh N, Bahrami A, Atkin SL, Sahebkar A. 2018. Role of the NLRP3 inflammasome in cancer. Mol. Cancer 17: 158.   DOI
30 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.   DOI
31 Ballestar E, Esteller M. 2008. Epigenetic gene regulation in cancer. Adv. Genet. 61: 247-267.   DOI
32 Hui AM, Makuuchi M, Li X. 1998. Cell cycle regulators and human hepatocarcinogenesis. Hepatogastroenterology 45: 1635-1642.
33 Diepenbruck M, Christofori G. 2016. Epithelial-mesenchymal transition (EMT) and metastasis: yes, no, maybe?. Curr. Opin. Cell Biol. 43: 7-13.   DOI
34 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.   DOI