Targeting protein palmitoylation decreases palmitate‑induced sphere formation of human liver cancer cells

Chong,Lee‑Won; Tsai,Chia‑Ling; Yang,Kou‑Ching; Liao,Chen‑Chung; Hsu,Yi‑Chao

doi:10.3892/mmr.2020.11172

Targeting protein palmitoylation decreases palmitate‑induced sphere formation of human liver cancer cells

Authors:
- Lee‑Won Chong
- Chia‑Ling Tsai
- Kou‑Ching Yang
- Chen‑Chung Liao
- Yi‑Chao Hsu
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Affiliations: Division of Hepatology and Gastroenterology, Department of Internal Medicine, Shin Kong Wu Ho Su Memorial Hospital, Taipei, Taiwan, R.O.C., Institute of Biomedical Sciences, Mackay Medical College, New Taipei City, Taipei, Taiwan, R.O.C., Proteomics Research Center, National Yang‑Ming University, Taipei, Taiwan, R.O.C.
Published online on: May 22, 2020 https://doi.org/10.3892/mmr.2020.11172
Pages: 939-947
Copyright: © Chong et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Although non‑alcoholic fatty liver disease (NAFLD) is considered a benign disorder, hepatic steatosis has been proposed to be involved in the tumorigenesis of liver cancer. However, the underlying mechanism for carcinogenesis in fatty liver diseases remains unclear. Cancer stem cells (CSCs) have been hypothesized to serve a key role in tumorigenesis. Tumor formation begins with a subset of heterogeneous cells that share properties with stem cells, such as self‑renewal and undifferentiated properties. Our previous study reported that the saturated fatty acid palmitate (PA) significantly enhanced the CSC properties of the HepG2 human liver cancer cell line; however, its underlying mechanisms are unknown. In the present study, a proteomic approach was used to investigate the palmitoylation of proteins in HepG2 CSCs. CSC behavior was induced in HepG2 cells via 200 µM PA. Proteomic analysis was performed to identify post‑transcriptional modifications of proteins in HepG2 CSCs in response to PA treatment. The present study identified proteins modified by palmitoylation in HepG2 CSC spheres formed following PA treatment. It was therefore hypothesized that palmitoylation may be crucial for CSC sphere formation. Furthermore, the present study demonstrated that two palmitoylation inhibitors, tunicamycin (5, 10 and 25 µg/ml) and 2‑bromohexadecanoic acid (25, 50 and 150 µM), significantly decreased CSC sphere formation without affecting cell viability. An association was identified between sphere formation capacity and tumor‑initiating capacity of CSCs. The results of the present study demonstrated that protein palmitoylation may influence the PA‑induced CSC tumor‑initiating capacity, and that the inhibition of palmitoylation may be a suitable chemopreventive strategy for treating patients with NAFLD.

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1	Younossi ZM, Koenig AB, Abdelatif D, Fazel Y, Henry L and Wymer M: Global epidemiology of nonalcoholic fatty liver disease-Meta-analytic assessment of prevalence, incidence, and outcomes. Hepatology. 64:73–84. 2016. View Article : Google Scholar : PubMed/NCBI
2	Younossi Z, Tacke F, Arrese M, Chander Sharma B, Mostafa I, Bugianesi E, Wai-Sun Wong V, Yilmaz Y, George J, Fan J and Vos MB: Global perspectives on non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. Hepatology. 69:2672–2682. 2019. View Article : Google Scholar : PubMed/NCBI
3	Angulo P and Lindor KD: Treatment of non-alcoholic steatohepatitis. Best Pract Res Clin Gastroenterol. 16:797–810. 2002. View Article : Google Scholar : PubMed/NCBI
4	Malaguarnera M, Di Rosa M, Nicoletti F and Malaguarnera L: Molecular mechanisms involved in NAFLD progression. J Mol Med (Berl). 87:679–695. 2009. View Article : Google Scholar : PubMed/NCBI
5	Leamy AK, Egnatchik RA and Young JD: Molecular mechanisms and the role of saturated fatty acids in the progression of non-alcoholic fatty liver disease. Prog Lipid Res. 52:165–174. 2013. View Article : Google Scholar : PubMed/NCBI
6	Cao L, Zhou Y, Zhai B, Liao J, Xu W, Zhang R, Li J, Zhang Y, Chen L, Qian H, et al: Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines. BMC Gastroenterol. 11:712011. View Article : Google Scholar : PubMed/NCBI
7	Huang P, Qiu J, Li B, Hong J, Lu C, Wang L, Wang J, Hu Y, Jia W and Yuan Y: Role of Sox2 and Oct4 in predicting survival of hepatocellular carcinoma patients after hepatectomy. Clin Biochem. 44:582–589. 2011. View Article : Google Scholar : PubMed/NCBI
8	Sun C, Sun L, Li Y, Kang X, Zhang S and Liu Y: Sox2 expression predicts poor survival of hepatocellular carcinoma patients and it promotes liver cancer cell invasion by activating Slug. Med Oncol. 30:5032013. View Article : Google Scholar : PubMed/NCBI
9	Yin X, Li YW, Zhang BH, Ren ZG, Qiu SJ, Yi Y and Fan J: Coexpression of stemness factors Oct4 and Nanog predict liver resection. Ann Surg Oncol. 19:2877–2887. 2012. View Article : Google Scholar : PubMed/NCBI
10	Ma XL, Sun YF, Wang BL, Shen MN, Zhou Y, Chen JW, Hu B, Gong ZJ, Zhang X, Cao Y, et al: Sphere-forming culture enriches liver cancer stem cells and reveals Stearoyl-CoA desaturase 1 as a potential therapeutic target. BMC Cancer. 19:7602019. View Article : Google Scholar : PubMed/NCBI
11	Stafman LL, Williams AP, Garner EF, Aye JM, Stewart JE, Yoon KJ, Whelan K and Beierle EA: Targeting PIM kinases affects maintenance of CD133 tumor cell population in hepatoblastoma. Transl Oncol. 12:200–208. 2019. View Article : Google Scholar : PubMed/NCBI
12	Maehara O, Ohnishi S, Asano A, Suda G, Natsuizaka M, Nakagawa K, Kobayashi M, Sakamoto N and Takeda H: Metformin regulates the expression of CD133 through the AMPK-CEBPβ pathway in hepatocellular carcinoma cell lines. Neoplasia. 21:545–556. 2019. View Article : Google Scholar : PubMed/NCBI
13	Chen WC, Chang YS, Hsu HP, Yen MC, Huang HL, Cho CY, Wang CY, Weng TY, Lai PT, Chen CS, et al: Therapeutics targeting CD90-integrin-AMPK-CD133 signal axis in liver cancer. Oncotarget. 6:42923–42937. 2015. View Article : Google Scholar : PubMed/NCBI
14	He J, Liu Y, Zhu T, Zhu J, Dimeco F, Vescovi AL, Heth JA, Muraszko KM, Fan X and Lubman DM: CD90 is identified as a candidate marker for cancer stem cells in primary high-grade gliomas using tissue microarrays. Mol Cell Proteomics. 11:M111.010744. 2012. View Article : Google Scholar
15	Zhang K, Che S, Pan C, Su Z, Zheng S, Yang S, Zhang H, Li W, Wang W and Liu J: The SHH/Gli axis regulates CD90-mediated liver cancer stem cell function by activating the IL6/JAK2 pathway. J Cell Mol Med. 22:3679–3690. 2018. View Article : Google Scholar : PubMed/NCBI
16	Wei S, Liu K, He Q, Gao Y and Shen L: PES1 is regulated by CD44 in liver cancer stem cells via miR-105-5p. FEBS Lett. 593:1777–1786. 2019. View Article : Google Scholar : PubMed/NCBI
17	Yamashita T, Honda M, Nakamoto Y, Baba M, Nio K, Hara Y, Zeng SS, Hayashi T, Kondo M, Takatori H, et al: Discrete nature of EpCAM+ and CD90+ cancer stem cells in human hepatocellular carcinoma. Hepatology. 57:1484–1497. 2013. View Article : Google Scholar : PubMed/NCBI
18	Christ B, Stock P and Dollinger MM: CD13: Waving the flag for a novel cancer stem cell target. Hepatology. 53:1388–1390. 2011. View Article : Google Scholar : PubMed/NCBI
19	Reynolds BA and Weiss S: Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell. Dev Biol. 175:1–13. 1996. View Article : Google Scholar : PubMed/NCBI
20	Dontu G, Abdallah WM, Foley JM, Jackson KW, Clarke MF, Kawamura MJ and Wicha MS: In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev. 17:1253–1270. 2003. View Article : Google Scholar : PubMed/NCBI
21	Suzuki A, Oyama K, Fukao K, Nakauchi H and Taniguchi H: Establishment of clonal colony-forming assay system for pancreatic stem/progenitor cells. Cell Transplant. 11:451–453. 2002. View Article : Google Scholar : PubMed/NCBI
22	Shi X, Gipp J and Bushman W: Anchorage-independent culture maintains prostate stem cells. Dev Biol. 312:396–406. 2007. View Article : Google Scholar : PubMed/NCBI
23	Ponti D, Costa A, Zaffaroni N, Pratesi G, Petrangolini G, Coradini D, Pilotti S, Pierotti MA and Daidone MG: Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res. 65:5506–5511. 2005. View Article : Google Scholar : PubMed/NCBI
24	Gou S, Liu T, Wang C, Yin T, Li K, Yang M and Zhou J: Establishment of clonal colony-forming assay for propagation of pancreatic cancer cells with stem cell properties. Pancreas. 34:429–435. 2007. View Article : Google Scholar : PubMed/NCBI
25	Rappa G, Mercapide J, Anzanello F, Prasmickaite L, Xi Y, Ju J, Fodstad O and Lorico A: Growth of cancer cell lines under stem cell-like conditions has the potential to unveil therapeutic targets. Exp Cell Res. 314:2110–2122. 2008. View Article : Google Scholar : PubMed/NCBI
26	Wobser H, Dorn C, Weiss TS, Amann T, Bollheimer C, Büttner R, Schölmerich J and Hellerbrand C: Lipid accumulation in hepatocytes induces fibrogenic activation of hepatic stellate cells. Cell Res. 19:996–1005. 2009. View Article : Google Scholar : PubMed/NCBI
27	Chong LW, Chou RH, Liao CC, Lee TF, Lin Y, Yang KC and Hsu YC: Saturated fatty acid induces cancer stem cell-like properties in human hepatoma cells. Cell Mol Biol (Noisy-le-Grand). 61:85–91. 2015.PubMed/NCBI
28	Anderson AM and Ragan MA: Palmitoylation: A protein S-acylation with implications for breast cancer. NPJ Breast Cancer. 2:160282016. View Article : Google Scholar : PubMed/NCBI
29	Yang MH, Liao CC, Hung JH, Lai XT, Yen CH and Chen YA: Utilizing proteomic approach to identify nuclear translocation related serine kinase phosphorylation site of GNMT as downstream effector for benzo[a]pyrene. J Food Drug Anal. 27:603–609. 2019. View Article : Google Scholar : PubMed/NCBI
30	Chong LW, Hsu YC, Lee TF, Lin Y, Chiu YT, Yang KC, Wu JC and Huang YT: Fluvastatin attenuates hepatic steatosis-induced fibrogenesis in rats through inhibiting paracrine effect of hepatocyte on hepatic stellate cells. BMC Gastroenterol. 15:222015. View Article : Google Scholar : PubMed/NCBI
31	Alessio N, Del Gaudio S, Capasso S, Di Bernardo G, Cappabianca S, Cipollaro M, Peluso G and Galderisi U: Low dose radiation induced senescence of human mesenchymal stromal cells and impaired the autophagy process. Oncotarget. 6:8155–8166. 2015. View Article : Google Scholar : PubMed/NCBI
32	Buckley BJ and Whorton AR: Tunicamycin increases intracellular calcium levels in bovine aortic endothelial cells. Am J Physiol. 273:C1298–C1305. 1997. View Article : Google Scholar : PubMed/NCBI
33	Sobocińska J, Roszczenko-Jasińska P, Zaręba-Kozioł M, Hromada-Judycka A, Matveichuk OV, Traczyk G, Łukasiuk K and Kwiatkowska K: Lipopolysaccharide upregulates palmitoylated enzymes of the phosphatidylinositol cycle: An insight from proteomic studies. Mol Cell Proteomics. 17:233–254. 2018. View Article : Google Scholar : PubMed/NCBI
34	Patterson SI and Skene JH: Novel inhibitory action of tunicamycin homologues suggests a role for dynamic protein fatty acylation in growth cone-mediated neurite extension. J Cell Biol. 124:521–536. 1994. View Article : Google Scholar : PubMed/NCBI