Co-culture of hepatoma cells with hepatocytic precursor (stem-like) cells inhibits tumor cell growth and invasion by downregulating Akt/NF-κB expression

Sui,Cheng‑Jun; Xu,Miao; Li,Wei‑Qing; Yang,Jia‑Mei; Yan,Hong‑Zhu; Liu,Hui‑Min; Xia,Chun‑Yan; Yu,Hong‑Yu

doi:10.3892/ol.2016.5128

Co-culture of hepatoma cells with hepatocytic precursor (stem-like) cells inhibits tumor cell growth and invasion by downregulating Akt/NF-κB expression

Authors:
- Cheng‑Jun Sui
- Miao Xu
- Wei‑Qing Li
- Jia‑Mei Yang
- Hong‑Zhu Yan
- Hui‑Min Liu
- Chun‑Yan Xia
- Hong‑Yu Yu
View Affiliations

Affiliations: Department of Special Medical Care Unit I and Liver Transplantation, The Eastern Hepatobiliary Surgery Hospital, Shanghai 200438, P.R. China, Department of Geratology, Changhai Hospital, Shanghai 200438, P.R. China, Department of Pathology, Changzheng Hospital, The Second Military Medical University, Shanghai 200003, P.R. China
Published online on: September 13, 2016 https://doi.org/10.3892/ol.2016.5128
Pages: 4054-4060

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hepatocytic stem cells (HSCs) have inhibitory effects on hepatocarcinoma cells. The present study investigated the effects of HSC activity in hepatocarcinoma cells in vitro. A Transwell co‑culture system of hepatocytic precursor (stem‑like) WB‑F344 cells and hepatoma CBRH‑7919 cells was used to assess HSC activity in metastasized hepatoma cells in vitro. Nude mouse xenografts were used to assess HSC activity in vivo. Co‑culture of hepatoma CBRH‑7919 cells with WB‑F344 cells suppressed the growth and colony formation, tumor cell migration and invasion capacity of CBRH‑7919 cells. The nude mouse xenograft assay demonstrated that the xenograft size of CBRH‑7919 cells following co‑culture with WB‑F344 cells was significantly smaller compared with that of control cells. Furthermore, the expression levels of the epithelial markers E‑cadherin and β‑catenin were downregulated, while the mesenchymal markers α‑SMA and vimentin were upregulated. Co‑culture of CBRH‑7919 cells with WB‑F344 cells downregulated NF‑κB and phospho‑Akt expression. In conclusion, hepatocytic precursor (stem‑like) WB‑F344 cells inhibited the growth, colony formation and invasion capacity of metastasized hepatoma CBRH‑7919 cells in vitro and in vivo by downregulating Akt/NF‑κB signaling.

View Figures

View References

1	He J, Gu D, Wu X, et al: Major causes of death among men and women in China. N Engl J Med. 353:1124–1134. 2005. View Article : Google Scholar : PubMed/NCBI
2	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar : PubMed/NCBI
3	Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, et al: Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 359:378–390. 2008. View Article : Google Scholar : PubMed/NCBI
4	Nakamizo A, Marini F, Amano T, Khan A, Studeny M, Gumin J, Chen J, Hentschel S, Vecil G, Dembinski J, et al: Human bone marrow-derived mesenchymal stem cells in the treatment of gliomas. Cancer Res. 65:3307–3318. 2005.PubMed/NCBI
5	Aboody KS, Brown A, Rainov NG, Bower KA, Liu S, Yang W, Small JE, Herrlinger U, Ourednik V, Black PM, et al: Neural stem cells display extensive tropism for pathology in adult brain: Evidence from intracranial gliomas. Proc Natl Acad Sci U S A. 97:12846–12851. 2000. View Article : Google Scholar : PubMed/NCBI
6	Chen X, Lin X, Zhao J, Shi W, Zhang H, Wang Y, Kan B, Du L, Wang B, Wei Y, et al: A tumor-selective biotherapy with prolonged impact on established metastases based on cytokine gene-engineered MSCs. Mol Ther. 16:749–756. 2008. View Article : Google Scholar : PubMed/NCBI
7	Uchibori R, Okada T, Ito T, Urabe M, Mizukami H, Kume A and Ozawa K: Retroviral vector-producing mesenchymal stem cells for targeted suicide cancer gene therapy. J Gene Med. 11:373–381. 2009. View Article : Google Scholar : PubMed/NCBI
8	Ren C, Kumar S, Chanda D, Chen J, Mountz JD and Ponnazhagan S: Therapeutic potential of mesenchymal stem cells producing interferon-alpha in a mouse melanoma lung metastasis model. Stem Cells. 26:2332–2338. 2008. View Article : Google Scholar : PubMed/NCBI
9	Nakamura K, Ito Y, Kawano Y, Kurozumi K, Kobune M, Tsuda H, Bizen A, Honmou O, Niitsu Y and Hamada H: Antitumor effect of genetically engineered mesenchymal stem cells in a rat glioma model. Gene Ther. 11:1155–1164. 2004. View Article : Google Scholar : PubMed/NCBI
10	Zhong XG, He S, Yin W, Deng JY and Cheng B: Selective tropism of liver stem cells to hepatocellular carcinoma in vivo. World J Gastroenterol. 13:3886–3891. 2007. View Article : Google Scholar : PubMed/NCBI
11	Chen XC, Wang R, Zhao X, Wei YQ, Hu M, Wang YS, Zhang XW, Zhang R, Zhang L, Yao B, et al: Prophylaxis against carcinogenesis in three kinds of unestablished tumor models via IL12-gene-engineered MSCs. Carcinogenesis. 27:2434–2441. 2006. View Article : Google Scholar : PubMed/NCBI
12	Gao Y, Yao A, Zhang W, et al: Human mesenchymal stem cells overexpressing pigment epithelium-derived factor inhibit hepatocellular carcinoma in nude mice. Oncogene. 29:2784–2794. 2010. View Article : Google Scholar : PubMed/NCBI
13	Aquino JB, Bolontrade MF, García MG, Podhajcer OL and Mazzolini G: Mesenchymal stem cells as therapeutic tools and gene carriers in liver fibrosis and hepatocellular carcinoma. Gene Ther. 17:692–708. 2010. View Article : Google Scholar : PubMed/NCBI
14	Qiao L, Xu Z, Zhao T, et al: Suppression of tumorigenesis by human mesenchymal stem cells in a hepatoma model. Cell Res. 18:500–507. 2008. View Article : Google Scholar : PubMed/NCBI
15	Qiao L, Zhao TJ, Wang FZ, Shan CL, Ye LH and Zhang XD: NF-kappaB downregulation may be involved the depression of tumor cell proliferation mediated by human mesenchymal stem cells. Acta Pharmacol Sin. 29:333–340. 2008. View Article : Google Scholar : PubMed/NCBI
16	Campos B, Wan F, Farhadi M, Ernst A, Zeppernick F, Tagscherer KE, Ahmadi R, Lohr J, Dictus C, Gdynia G, et al: Differentiation therapy exerts antitumor effects on stem-like glioma cells. Clin Cancer Res. 16:2715–2728. 2010. View Article : Google Scholar : PubMed/NCBI
17	Omar HA, Sargeant AM, Weng JR, Wang D, Kulp SK, Patel T and Chen CS: Targeting of the Akt-nuclear factor-kappaB signaling network by [1-(4-chloro-3-nitrobenzenesulfonyl)-1H-indol-3-yl]-methanol (OSU-A9), a novel indole-3-carbinol derivative, in a mouse model of hepatocellular carcinoma. Mol Pharmacol. 76:957–968. 2009. View Article : Google Scholar : PubMed/NCBI
18	Wu LF, Li GP, Su JD, Pu ZJ, Feng JL, Ye YQ and Wei BL: Involvement of NF-kappaB activation in the apoptosis induced by extracellular adenosine in human hepatocellular carcinoma HepG2 cells. Biochem Cell Biol. 88:705–714. 2010. View Article : Google Scholar : PubMed/NCBI
19	Kim MO, Moon DO, Kang CH, Kwon TK, Choi YH and Kim GY: beta-Ionone enhances TRAIL-induced apoptosis in hepatocellular carcinoma cells through Sp1-dependent upregulation of DR5 and downregulation of NF-kappaB activity. Mol Cancer Ther. 9:833–843. 2010. View Article : Google Scholar : PubMed/NCBI
20	Ren K, Zhang W, Shi Y and Gong J: Pim-2 activates API-5 to inhibit the apoptosis of hepatocellular carcinoma cells through NF-kappaB pathway. Pathol Oncol Res. 16:229–237. 2010. View Article : Google Scholar : PubMed/NCBI
21	Cho HA, Park IS, Kim TW, Oh YK, Yang KS and Kim JS: Suppression of hepatitis B virus-derived human hepatocellular carcinoma by NF-kappaB-inducing kinase-specific siRNA using liver-targeting liposomes. Arch Pharm Res. 32:1077–1086. 2009. View Article : Google Scholar : PubMed/NCBI
22	Fei HR, Chen G, Wang JM and Wang FZ: Perifosine induces cell cycle arrest and apoptosis in human hepatocellular carcinoma cell lines by blockade of Akt phosphorylation. Cytotechnology. 62:449–460. 2010. View Article : Google Scholar : PubMed/NCBI
23	Buontempo F, Ersahin T, Missiroli S, Senturk S, Etro D, Ozturk M, Capitani S, Cetin-Atalay R and Neri ML: Inhibition of Akt signaling in hepatoma cells induces apoptotic cell death independent of Akt activation status. Invest New Drugs. 29:1303–1313. 2011. View Article : Google Scholar : PubMed/NCBI
24	Nair AS, Shishodia S, Ahn KS, Kunnumakkara AB, Sethi G and Aggarwal BB: Deguelin, an Akt inhibitor, suppresses IkappaBalpha kinase activation leading to suppression of NF-kappaB-regulated gene expression, potentiation of apoptosis and inhibition of cellular invasion. J Immunol. 177:5612–5622. 2006. View Article : Google Scholar : PubMed/NCBI
25	Ozes ON, Mayo LD, Gustin JA, Pfeffer SR, Pfeffer LM and Donner DB: NF-kappaB activation by tumour necrosis factor requires the Akt serine-threonine kinase. Nature. 401:82–85. 1999. View Article : Google Scholar : PubMed/NCBI
26	Tsao MS, Smith JD, Nelson KG and Grisham JW: A diploid epithelial cell line from normal adult rat liver with phenotypic properties of 'oval' cells. Exp Cell Res. 154:38–52. 1984. View Article : Google Scholar : PubMed/NCBI
27	Huber MA, Kraut N and Beug H: Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol. 17:548–558. 2005. View Article : Google Scholar : PubMed/NCBI
28	Kang Y and Massagué J: Epithelial-mesenchymal transitions: Twist in development and metastasis. Cell. 118:277–279. 2004. View Article : Google Scholar : PubMed/NCBI
29	Lin HH, Chen JH, Chou FP and Wang CJ: Protocatechuic acid inhibits cancer cell metastasis involving the down-regulation of Ras/Akt/NF-kappaB pathway and MMP-2 production by targeting RhoB activation. Br J Pharmacol. 162:237–254. 2011. View Article : Google Scholar : PubMed/NCBI
30	Park YP, Choi SC, Kim BY, Kim JT, Song EY, Kang SH, Yoon DY, Paik SG, Kim KD, Kim JW and Lee HG: Induction of Mac-2BP by nerve growth factor is regulated by the PI3K/Akt/NF-kappaB-dependent pathway in the HEK293 cell line. BMB Rep. 41:784–789. 2008. View Article : Google Scholar : PubMed/NCBI
31	Yuki K, Hirohashi S, Sakamoto M, Kanai T and Shimosato Y: Growth and spread of hepatocellular carcinoma. A review of 240 consecutive autopsy cases. Cancer. 66:2174–2179. 1990. View Article : Google Scholar : PubMed/NCBI
32	Genda T, Sakamoto M, Ichida T, Asakura H, Kojiro M, Narumiya S and Hirohashi S: Cell motility mediated by rho and Rho-associated protein kinase plays a critical role in intrahepatocytic metastasis of human hepatocellular carcinoma. Hepatology. 30:1027–1036. 1999. View Article : Google Scholar : PubMed/NCBI
33	Li L and Neaves WB: Normal stem cells and cancer stem cells: The niche matters. Cancer Res. 66:4553–4557. 2006. View Article : Google Scholar : PubMed/NCBI
34	Li GC, Ye QH, Xue YH, Sun HJ, Zhou HJ, Ren N, Jia HL, Shi J, Wu JC, Dai C, et al: Human mesenchymal stem cells inhibit metastasis of a hepatocellular carcinoma model using the MHCC97-H cell line. Cancer Sci. 101:2546–2553. 2010. View Article : Google Scholar : PubMed/NCBI
35	Karin M, Cao Y, Greten FR and Li ZW: NF-kappaB in cancer: From innocent bystander to major culprit. Nat Rev Cancer. 2:301–310. 2002. View Article : Google Scholar : PubMed/NCBI
36	Haefner B: NF-kappaB: Arresting a major culprit in cancer. Drug Discov Today. 7:653–663. 2002. View Article : Google Scholar : PubMed/NCBI
37	Testa JR and Bellacosa A: AKT plays a central role in tumorigenesis. Proc Natl Acad Sci U S A. 98:10983–10985. 2001. View Article : Google Scholar : PubMed/NCBI
38	Cheng JQ, Lindsley CW, Cheng GZ, Yang H and Nicosia SV: The Akt/PKB pathway: Molecular target for cancer drug discovery. Oncogene. 24:7482–7492. 2005. View Article : Google Scholar : PubMed/NCBI
39	Wendel HG, De Stanchina E, Fridman JS, Malina A, Ray S, Kogan S, Cordon-Cardo C, Pelletier J and Lowe SW: Survival signalling by Akt and eIF4E in oncogenesis and cancer therapy. Nature. 428:332–337. 2004. View Article : Google Scholar : PubMed/NCBI
40	Clark AS, West K, Streicher S and Dennis PA: Constitutive and inducible Akt activity promotes resistance to chemotherapy, trastuzumab, or tamoxifen in breast cancer cells. Mol Cancer Ther. 1:707–717. 2002.PubMed/NCBI
41	Li Y, Ahmed F, Ali S, Philip PA, Kucuk O and Sarkar FH: Inactivation of nuclear factor kappaB by soy isoflavone genistein contributes to increased apoptosis induced by chemotherapeutic agents in human cancer cells. Cancer Res. 65:6934–6942. 2005. View Article : Google Scholar : PubMed/NCBI
42	Konrath F, Witt J, Sauter T and Kulms D: Identification of new IκBα complexes by an iterative experimental and mathematical modeling approach. PLoS Comput Biol. 10:e10035282014. View Article : Google Scholar : PubMed/NCBI