Downregulation of hepatic lipase is associated with decreased CD133 expression and clone formation in HepG2 cells

Liu,Xuehua; Zuo,Junhua; Fang,Yuan; Wen,Jing; Deng,Feihong; Zhong,Hui; Jiang,Bo; Wang,Jide; Nie,Biao

doi:10.3892/ijmm.2018.3756

Downregulation of hepatic lipase is associated with decreased CD133 expression and clone formation in HepG2 cells

Authors:
- Xuehua Liu
- Junhua Zuo
- Yuan Fang
- Jing Wen
- Feihong Deng
- Hui Zhong
- Bo Jiang
- Jide Wang
- Biao Nie
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Affiliations: Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Gastroenterology, The Fifth Affiliated Hospital of Southern Medical University, Guangzhou, Guangdong 510900, P.R. China, Department of Gastroenterology, Beijing Tsinghua Changgung Hospital, Tsinghua University, Beijing 102218, P.R. China, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510630, P.R. China
Published online on: July 4, 2018 https://doi.org/10.3892/ijmm.2018.3756
Pages: 2137-2144

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Abstract

The drug resistance of cancer remains a major obstacle to successful chemotherapy. New strategies for improving chemotherapeutic efficacy are urgently required. Recent studies have indicated that LIPC plays a role in promoting the liver metastasis of colorectal cancer. In the present study, we aimed to investigate the effects of LIPC on theproliferation and clone formation of colorectal cancer-derived cells, and chemoresistance in hepatoblastoma-derived HepG2 cells. The activity and expression of LIPC were determined in the cell lines by RT-qPCR and western blot analysis. HepG2 cells in which LIPC was knocked down by LIPC short hairpin RNA (shRNA) and control cells [shRNA control (shCON)] were established and analyzed for cell proliferation and colony formation rates. FACS analysis was used to explore the association between LIPC and the tumor-derived cell biomarker, CD133, and the percentages of CD133-positive cells were assessed by FACS. Additionally, shLIPC- and shCON-transfected cells were treated with various concentrations of doxorubicin and 5-floxuridine (5-FU), and cell viability was determined by MTT assay. mRNA levels in the shLIPC- and shCON-transfected cells were compared by cDNA microarray and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The results revealed that the HepG2 cells exhibited a relatively higher LIPC activity and expression levels compared to the other colon cancer cell lines. The downregulation of LIPC in the HepG2 cells was associated with the decreased expression of CD133, decreased cell proliferation and colony formation, as well as increased resistance to chemotherapy. KEGG analysis of the cDNA microarray data revealed increased levels in the cell adhesion molecule (CAM) pathway, including CLDN10 and CLDN1, indicating that CAMs may play a role in LIPC-mediated tumor progression. The present findings indicate a potential role of LIPC as a promising therapeutic target in cancer.

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1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
2	Asghar U and Meyer T: Are there opportunities for chemotherapy in the treatment of hepatocellular cancer? J Hepatol. 56:686–695. 2012. View Article : Google Scholar
3	Sukowati CH, Rosso N, Crocè LS and Tiribelli C: Hepatic cancer stem cells and drug resistance: Relevance in targeted therapies for hepatocellular carcinoma. World J Hepatol. 2:114–126. 2010. View Article : Google Scholar : PubMed/NCBI
4	Wong H and Schotz MC: The lipase gene family. J Lipid Res. 43:993–999. 2002. View Article : Google Scholar : PubMed/NCBI
5	Santamarina-Fojo S, González-Navarro H, Freeman L, Wagner E and Nong Z: Hepatic lipase, lipoprotein metabolism, and atherogenesis. Arterioscler Thromb Vasc Biol. 24:1750–1754. 2004. View Article : Google Scholar : PubMed/NCBI
6	Nomura DK, Long JZ, Niessen S, Hoover HS, Ng SW and Cravatt BF: Monoacylglycerol lipase regulates a fatty acid network that promotes cancer pathogenesis. Cell. 140:49–61. 2010. View Article : Google Scholar : PubMed/NCBI
7	Ding Q, Chang CJ, Xie X, Xia W, Yang JY, Wang SC, Wang Y, Xia J, Chen L, Cai C, et al: APOBEC3G promotes liver metastasis in an orthotopic mouse model of colorectal cancer and predicts human hepatic metastasis. J Clin Invest. 121:4526–4536. 2011. View Article : Google Scholar : PubMed/NCBI
8	Galluzzi L, Goubar A, Olaussen KA, Vitale I, Senovilla L, Michels J, Robin A, Dorvault N, Besse B, Validire P, et al: Prognostic value of LIPC in non-small cell lung carcinoma. Cell Cycle. 12:647–654. 2013. View Article : Google Scholar : PubMed/NCBI
9	Ma S, Lee TK, Zheng BJ, Chan KW and Guan XY: CD133⁺ HCC cancer stem cells confer chemoresistance by preferential expression of the Akt/PKB survival pathway. Oncogene. 27:1749–1758. 2008. View Article : Google Scholar
10	Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, Castedo M and Kroemer G: Molecular mechanisms of cisplatin resistance. Oncogene. 31:1869–1883. 2012. View Article : Google Scholar
11	Livak and Schmittgen: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
12	Franken NA, Rodermond HM, Stap J, Haveman J and van Bree C: Clonogenic assay of cells in vitro. Nat Protoc. 1:2315–2319. 2006. View Article : Google Scholar
13	Stein W, Bohner J and Bahlinger M: Macro lipase - a new member of the family of immunoglobulin-linked enzymes. J Clin Chem Clin Biochem. 25:837–843. 1987.PubMed/NCBI
14	Muñoz-Perez M, Sarrion-Pelous D, Jimenez-Jimenez J, Martinez-Montiel P and Gallego-Valdes M: Chronic increased serum lipase in a patient with suspected pancreatic adenocarcinoma. Clin Chem. 43:191–193. 1997.PubMed/NCBI
15	Diani G, Poma G, Novazzi F, Zanirato S, Porta C, Moroni M, Melzi d’ Eril GV and Moratti R: Increased serum lipase with associated normoamylasemia in cancer patients. Clin Chem. 44:1043–1045. 1998.PubMed/NCBI
16	Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J and Dirks PB: Identification of a cancer stem cell in human brain tumors. Cancer Res. 63:5821–5828. 2003.PubMed/NCBI
17	Fang D, Nguyen TK, Leishear K, Finko R, Kulp AN, Hotz S, Van Belle PA, Xu X, Elder DE and Herlyn M: A tumorigenic subpopulation with stem cell properties in melanomas. Cancer Res. 65:9328–9337. 2005. View Article : Google Scholar : PubMed/NCBI
18	Collins AT, Berry PA, Hyde C, Stower MJ and Maitland NJ: Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res. 65:10946–10951. 2005. View Article : Google Scholar : PubMed/NCBI
19	Miki J, Furusato B, Li H, Gu Y, Takahashi H, Egawa S, Sesterhenn IA, McLeod DG, Srivastava S and Rhim JS: Identification of putative stem cell markers, CD133 and CXCR4, in hTERT-immortalized primary nonmalignant and malignant tumor-derived human prostate epithelial cell lines and in prostate cancer specimens. Cancer Res. 67:3153–3161. 2007. View Article : Google Scholar : PubMed/NCBI
20	O’Brien CA, Pollett A, Gallinger S and Dick JE: A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature. 445:106–110. 2007. View Article : Google Scholar
21	Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C and De Maria R: Identification and expansion of human colon-cancer-initiating cells. Nature. 445:111–115. 2007. View Article : Google Scholar
22	Suetsugu A, Nagaki M, Aoki H, Motohashi T, Kunisada T and Moriwaki H: Characterization of CD133+ hepatocellular carcinoma cells as cancer stem/progenitor cells. Biochem Biophys Res Commun. 351:820–824. 2006. View Article : Google Scholar : PubMed/NCBI
23	Yin S, Li J, Hu C, Chen X, Yao M, Yan M, Jiang G, Ge C, Xie H, Wan D, et al: CD133 positive hepatocellular carcinoma cells possess high capacity for tumorigenicity. Int J Cancer. 120:1444–1450. 2007. View Article : Google Scholar : PubMed/NCBI
24	Bao S, Wu Q, Sathornsumetee S, Hao Y, Li Z, Hjelmeland AB, Shi Q, McLendon RE, Bigner DD and Rich JN: Stem cell-like glioma cells promote tumor angiogenesis through vascular endothelial growth factor. Cancer Res. 66:7843–7848. 2006. View Article : Google Scholar : PubMed/NCBI
25	Shmelkov SV, Butler JM, Hooper AT, Hormigo A, Kushner J, Milde T, St Clair R, Baljevic M, White I, Jin DK, et al: CD133 expression is not restricted to stem cells, and both CD133⁺ and CD133⁻ metastatic colon cancer cells initiate tumors. J Clin Invest. 118:2111–2120. 2008.PubMed/NCBI
26	Meng X, Li M, Wang X, Wang Y and Ma D: Both CD133⁺ and CD133⁻ subpopulations of A549 and H446 cells contain cancer-initiating cells. Cancer Sci. 100:1040–1046. 2009. View Article : Google Scholar : PubMed/NCBI
27	Jaksch M, Múnera J, Bajpai R, Terskikh A and Oshima RG: Cell cycle-dependent variation of a CD133 epitope in human embryonic stem cell, colon cancer, and melanoma cell lines. Cancer Res. 68:7882–7886. 2008. View Article : Google Scholar : PubMed/NCBI
28	Yang JD, Nakamura I and Roberts LR: The tumor microenvironment in hepatocellular carcinoma: Current status and therapeutic targets. Semin Cancer Biol. 21:35–43. 2011. View Article : Google Scholar :
29	Cheung ST, Leung KL, Ip YC, Chen X, Fong DY, Ng IO, Fan ST and So S: Claudin-10 expression level is associated with recurrence of primary hepatocellular carcinoma. Clin Cancer Res. 11:551–556. 2005.PubMed/NCBI
30	Ip YC, Cheung ST, Lee YT, Ho JC and Fan ST: Inhibition of hepatocellular carcinoma invasion by suppression of claudin-10 in HLE cells. Mol Cancer Ther. 6:2858–2867. 2007. View Article : Google Scholar : PubMed/NCBI
31	Suh Y, Yoon CH, Kim RK, Lim EJ, Oh YS, Hwang SG, An S, Yoon G, Gye MC, Yi JM, et al: Claudin-1 induces epithelial-mesenchymal transition through activation of the c-Abl-ERK signaling pathway in human liver cells. Oncogene. 32:4873–4882. 2013. View Article : Google Scholar