The effect and mechanism of bufalin on regulating hepatocellular carcinoma cell invasion and metastasis via Wnt/β-catenin signaling pathway

Gai,Ji  Qin; Sheng,Xia; Qin,Jian  Min; Sun,Kang; Zhao,Wei; Ni,Lei

doi:10.3892/ijo.2015.3250

The effect and mechanism of bufalin on regulating hepatocellular carcinoma cell invasion and metastasis via Wnt/β-catenin signaling pathway

Authors:
- Ji Qin Gai
- Xia Sheng
- Jian Min Qin
- Kang Sun
- Wei Zhao
- Lei Ni
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Affiliations: Department of General Surgery, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, P.R. China, Department of Pathology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, P.R. China
Published online on: November 18, 2015 https://doi.org/10.3892/ijo.2015.3250
Pages: 338-348

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Abstract

Hepatocellular carcinoma (HCC) is a highly malignant tumor with an extremely poor prognosis. Our preliminary study indicated that bufalin could restrain the proliferation of human hepatoma BEL-7402 cells in a time- and dose-dependent manner. In the present study, the colony formation assay, the Transwell invasion assay, the western blot analysis and the immunofluorescence method were respectively used to investigate the effect and mechanism of bufalin against HCC cell invasion and metastasis. We found that: i) bufalin had significant inhibitory effect on the cell proliferation of BEL-7402 cells; ii) bufalin markedly inhibited the migration and invasion of BEL-7402 cells; iii) bufalin could suppress the phosphorylation of GSK-3β Ser9 site in BEL-7402 cells, decrease the expression of β-catenin, cyclin D1, metalloproteinases-7 (MMP-7) and cyclooxygenase-2 (COX-2) in the cytoplasm, and increase the expression of E-cadherin and β-catenin on the cell membrane; and iv) the expression of α-fetoprotein significantly decreased and the expression of albumin increased in BEL-7402 cells after bufalin was used. Our results indicate that: i) bufalin can regulate the expression of associated factors in Wnt/β-catenin signaling pathway of BEL-7402 cells through suppressing the phosphorylation of GSK-3β Ser9 site; ii) bufalin can strengthen intercellular E-cadherin/β-catenin complex to control epithelial-mesenchymal transition; and iii) bufalin can reverse the malignant phenotype and promote the differentiation and maturation by regulating the AFP and ALB expression in BEL-7402 cells. These are very important mechanisms of bufalin on the inhibition of the invasion and metastasis of HCC cells.

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1	Song MJ and Bae SH: Newer treatments for advanced hepatocellular carcinoma. Korean J Intern Med. 29:149–155. 2014. View Article : Google Scholar : PubMed/NCBI
2	Wang Y, Shen Z, Zhu Z, Han R and Huai M and Huai M: Clinical values of AFP, GPC3 mRNA in peripheral blood for prediction of hepatocellular carcinoma recurrence following OLT: AFP, GPC3 mRNA for prediction of HCC. Hepat Mon. 11:195–199. 2011.PubMed/NCBI
3	Zhao YJ, Ju Q and Li GC: Tumor markers for hepatocellular carcinoma. Mol Clin Oncol. 1:593–598. 2013.
4	Jo S and Shim HK: A patient who has survived for a long period with repeated radiotherapies for multifocal extrahepatic metastases from hepatocellular carcinoma. Radiat Oncol J. 31:267–272. 2013. View Article : Google Scholar
5	Lin S, Hoffmann K and Schemmer P: Treatment of hepatocellular carcinoma: A systematic review. Liver Cancer. 1:144–158. 2012. View Article : Google Scholar
6	Belghiti J and Fuks D: Liver resection and transplantation in hepatocellular carcinoma. Liver Cancer. 1:71–82. 2012. View Article : Google Scholar
7	Vitale A, Volk M and Cillo U: Transplant benefit for patients with hepatocellular carcinoma. World J Gastroenterol. 19:9183–9188. 2013. View Article : Google Scholar
8	Chan SC: Liver transplantation for hepatocellular carcinoma. Liver Cancer. 2:338–344. 2013. View Article : Google Scholar
9	Yin PH, Liu X, Qiu YY, Cai JF, Qin JM, Zhu HR and Li Q: Anti-tumor activity and apoptosis-regulation mechanisms of bufalin in various cancers: New hope for cancer patients. Asian Pac J Cancer Prev. 13:5339–5343. 2012. View Article : Google Scholar
10	Gu W, Liu L, Fang FF, Huang F, Cheng BB and Li B: Reversal effect of bufalin on multidrug resistance in human hepatocellular carcinoma BEL-7402/5-FU cells. Oncol Rep. 31:216–222. 2014.
11	Gao Y, Li HX, Xu LT, Wang P, Xu LY, Cohen L, Yang PY, Gu K and Meng ZQ: Bufalin enhances the anti-proliferative effect of sorafenib on human hepatocellular carcinoma cells through downregulation of ERK. Mol Biol Rep. 39:1683–1689. 2012. View Article : Google Scholar
12	Gai JQ, Qin JM and Fan YZ: The effect of bufalin on proliferation and invasion of human liver cancer cells. World Chin J Digestology. 22:1921–1927. 2014.
13	Pandurangan AK: Potential targets for prevention of colorectal cancer: A focus on PI3K/Akt/mTOR and Wnt pathways. Asian Pac J Cancer Prev. 14:2201–2205. 2013. View Article : Google Scholar : PubMed/NCBI
14	Xu MX, Zhao L, Deng C, Yang L, Wang Y, Guo T, Li L, Lin J and Zhang L: Curcumin suppresses proliferation and induces apoptosis of human hepatocellular carcinoma cells via the wnt signaling pathway. Int J Oncol. 43:1951–1959. 2013.PubMed/NCBI
15	Wang F, He L, Dai WQ, Xu YP, Wu D, Lin CL, Wu SM, Cheng P, Zhang Y, Shen M, et al: Salinomycin inhibits proliferation and induces apoptosis of human hepatocellular carcinoma cells in vitro and in vivo. PLoS One. 7:e506382012. View Article : Google Scholar
16	Thompson MD, Dar MJ and Monga SP: Pegylated interferon alpha targets Wnt signaling by inducing nuclear export of β-catenin. J Hepatol. 54:506–512. 2011. View Article : Google Scholar :
17	Cairns RA, Harris IS and Mak TW: Regulation of cancer cell metabolism. Nat Rev Cancer. 11:85–95. 2011. View Article : Google Scholar : PubMed/NCBI
18	Bachmann J, Raue A, Schilling M, Becker V, Timmer J and Klingmüller U: Predictive mathematical models of cancer signalling pathways. J Intern Med. 271:155–165. 2012. View Article : Google Scholar
19	Di Maio A, Setar L, Tiozzo S and De Tomaso AW: Wnt affects symmetry and morphogenesis during post-embryonic development in colonial chordates. Evodevo. 6:172015. View Article : Google Scholar : PubMed/NCBI
20	Sastre-Perona A and Santisteban P: Role of the wnt pathway in thyroid cancer. Front Endocrinol (Lausanne). 3:312012.
21	Saito-Diaz K, Chen TW, Wang X, Thorne CA, Wallace HA, Page-McCaw A and Lee E: The way Wnt works: Components and mechanism. Growth Factors. 31:1–31. 2013. View Article : Google Scholar :
22	Ren S, Johnson BG, Kida Y, Ip C, Davidson KC, Lin SL, Kobayashi A, Lang RA, Hadjantonakis AK, Moon RT, et al: LRP-6 is a coreceptor for multiple fibrogenic signaling pathways in pericytes and myofibroblasts that are inhibited by DKK-1. Proc Natl Acad Sci USA. 110:1440–1445. 2013. View Article : Google Scholar : PubMed/NCBI
23	Fuchs SY, Ougolkov AV, Spiegelman VS and Minamoto T: Oncogenic beta-catenin signaling networks in colorectal cancer. Cell Cycle. 4:1522–1539. 2005. View Article : Google Scholar : PubMed/NCBI
24	Nagasundaram N, Hailong Z, Jiming L, Karthick V, George Priya Doss C, Chirangib C and Luonan C: Analysing the effect of mutation on protein function and discovering potential inhibitors of CDK4: Molecular modelling and dynamics studies. PLoS One. 10:e01339692015. View Article : Google Scholar
25	Wu J, Guan X, Zhang K, Li YT, Bai P and Wu J: A/G polymorphism of matrix metalloproteinase 7 gene promoter region and cancer risk: A meta-analysis. Biomed Rep. 1:792–796. 2013.
26	Wu KK, Cheng HH and Chang TC: 5-methoxyindole metabolites of L-tryptophan: Control of COX-2 expression, inflammation and tumorigenesis. J Biomed Sci. 21:172014. View Article : Google Scholar : PubMed/NCBI
27	Lee MJ, Xu DY, Li H, Yu GR, Leem SH, Chu IS, Kim IH and Kim DG: Pro-oncogenic potential of NM23-H2 in hepatocellular carcinoma. Exp Mol Med. 44:214–224. 2012. View Article : Google Scholar :
28	Chen L, Li M, Li Q, Wang CJ and Xie SQ: DKK1 promotes hepatocellular carcinoma cell migration and invasion through β-catenin/MMP7 signaling pathway. Mol Cancer. 12:1572013. View Article : Google Scholar
29	Sui W, Zhang Y, Wang Z, Wang Z, Jia Q, Wu L and Zhang W: Antitumor effect of a selective COX-2 inhibitor, celecoxib, may be attributed to angiogenesis inhibition through modulating the PTEN/PI3K/Akt/HIF-1 pathway in an H₂₂ murine hepatocarcinoma model. Oncol Rep. 31:2252–2260. 2014.PubMed/NCBI
30	Wands JR and Kim M: WNT/β-catenin signaling and hepatocellular carcinoma. Hepatology. 60:452–454. 2014. View Article : Google Scholar : PubMed/NCBI
31	Somorjai IM and Martinez-Arias A: Wingless signalling alters the levels, subcellular distribution and dynamics of Armadillo and E-cadherin in third instar larval wing imaginal discs. PLoS One. 3:e28932008. View Article : Google Scholar : PubMed/NCBI
32	Liu W, Wang H, Wang Y, Li H and Ji L: Metabolic factors-triggered inflammatory response drives antidepressant effects of exercise in CUMS rats. Psychiatry Res. 228:257–264. 2015. View Article : Google Scholar : PubMed/NCBI
33	Wolfe A, Thomas A, Edwards G, Jaseja R, Guo GL and Apte U: Increased activation of the Wnt/β-catenin pathway in spontaneous hepatocellular carcinoma observed in farnesoid X receptor knockout mice. J Pharmacol Exp Ther. 338:12–21. 2011. View Article : Google Scholar : PubMed/NCBI
34	Hou KZ, Fu ZQ and Gong H: Chemokine ligand 20 enhances progression of hepatocellular carcinoma via epithelial-mesenchymal transition. World J Gastroenterol. 21:475–483. 2015. View Article : Google Scholar : PubMed/NCBI
35	Kim HJ, Choi WJ and Lee CH: Phosphorylation and reorganization of keratin networks: Implications for carcinogenesis and epithelial mesenchymal transition. Biomol Ther (Seoul). 23:301–312. 2015. View Article : Google Scholar
36	Wei X, Wang J, He J, Ma B and Chen J: Biological characteristics of CD133⁺ cancer stem cells derived from human laryngeal carcinoma cell line. Int J Clin Exp Med. 7:2453–2462. 2014.
37	David JM and Rajasekaran AK: Dishonorable discharge: The oncogenic roles of cleaved E-cadherin fragments. Cancer Res. 72:2917–2923. 2012. View Article : Google Scholar : PubMed/NCBI
38	Xia W, Ma X, Li X, Dong H, Yi J, Zeng W and Yang Z: miR-153 inhibits epithelial-to-mesenchymal transition in hepatocellular carcinoma by targeting Snail. Oncol Rep. 34:655–662. 2015.PubMed/NCBI
39	Liao ZJ, Guo YH, Zhao Z, Yao JT, Xu R and Nan KJ: Gemcitabine inhibits the micrometastasis of non-small cell lung cancer by targeting the EpCAM-positive circulating tumor cells via the HGF/cMET pathway. Int J Oncol. 45:651–658. 2014.PubMed/NCBI
40	Quan MF, Xiao LH, Liu ZH, Guo H, Ren KQ, Liu F, Cao JG and Deng XY: 8-bromo-7-methoxychrysin inhibits properties of liver cancer stem cells via downregulation of β-catenin. World J Gastroenterol. 19:7680–7695. 2013. View Article : Google Scholar
41	Hu Z and Zhao W: Novel insights into the molecular mechanisms of α-fetoprotein expression and malignant phenotypes of hepatocellular carcinoma. Cell Mol Immunol. 9:7–8. 2012. View Article : Google Scholar
42	Kim HA, Nam K, Lee M and Kim SW: Hypoxia/hepatoma dual specific suicide gene expression plasmid delivery using bio-reducible polymer for hepatocellular carcinoma therapy. J Control Release. 171:1–10. 2013. View Article : Google Scholar : PubMed/NCBI
43	Garcia-Martinez R, Caraceni P, Bernardi M, Gines P, Arroyo V and Jalan R: Albumin: Pathophysiologic basis of its role in the treatment of cirrhosis and its complications. Hepatology. 58:1836–1846. 2013. View Article : Google Scholar : PubMed/NCBI
44	You J, Raghunathan VK, Son KJ, Patel D, Haque A, Murphy CJ and Revzin A: Impact of nanotopography, heparin hydrogel microstructures, and encapsulated fibroblasts on phenotype of primary hepatocytes. ACS Appl Mater Interfaces. 7:12299–12308. 2015. View Article : Google Scholar :
45	Ohguchi S, Nakatsukasa H, Higashi T, Ashida K, Nouso K, Ishizaki M, Hino N, Kobayashi Y, Uematsu S and Tsuji T: Expression of alpha-fetoprotein and albumin genes in human hepatocellular carcinomas: Limitations in the application of the genes for targeting human hepatocellular carcinoma in gene therapy. Hepatology. 27:599–607. 1998. View Article : Google Scholar : PubMed/NCBI
46	Toro A, Ardiri A, Mannino M, Arcerito MC, Mannino G, Palermo F, Bertino G and Di Carlo I: Effect of pre- and post-treatment α-fetoprotein levels and tumor size on survival of patients with hepatocellular carcinoma treated by resection, transarterial chemoembolization or radiofrequency ablation: A retrospective study. BMC Surg. 14:402014. View Article : Google Scholar
47	Nojiri S and Joh T: Albumin suppresses human hepatocellular carcinoma proliferation and the cell cycle. Int J Mol Sci. 15:5163–5174. 2014. View Article : Google Scholar : PubMed/NCBI
48	Vernon AE and LaBonne C: Tumor metastasis: A new twist on epithelial-mesenchymal transitions. Curr Biol. 14:R719–R721. 2004. View Article : Google Scholar : PubMed/NCBI
49	Valastyan S and Weinberg RA: Tumor metastasis: Molecular insights and evolving paradigms. Cell. 147:275–292. 2011. View Article : Google Scholar : PubMed/NCBI
50	Yin P, Wang Y, Qiu Y, Hou L, Liu X, Qin J, Duan Y, Liu P, Qiu M and Li Q: Bufalin-loaded mPEG-PLGA-PLL-cRGD nanoparticles: Preparation, cellular uptake, tissue distribution, and anticancer activity. Int J Nanomed. 7:3961–3969. 2012.