Anticancer effect of 20(S)-ginsenoside Rh2 on HepG2 liver carcinoma cells: Activating GSK-3β and degrading β-catenin

Shi,Qingqiang; Shi,Xueping; Zuo,Gei; Xiong,Wei; Li,Haixing; Guo,Pei; Wang,Fen; Chen,Yi; Li,Jing; Chen,Di‑Long

doi:10.3892/or.2016.5033

Anticancer effect of 20(S)-ginsenoside Rh2 on HepG2 liver carcinoma cells: Activating GSK-3β and degrading β-catenin

Authors:
- Qingqiang Shi
- Xueping Shi
- Gei Zuo
- Wei Xiong
- Haixing Li
- Pei Guo
- Fen Wang
- Yi Chen
- Jing Li
- Di‑Long Chen
View Affiliations

Affiliations: Emergency Department of First People's Hospital of Chongqing New North Zone, Chongqing 401121, P.R. China, Laboratory of Stem Cell and Tissue Engineering, Department of Histology and Embryology, Chongqing Medical University, Chongqing 400016, P.R. China, Laboratory of Clinical Diagnostics, Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: August 24, 2016 https://doi.org/10.3892/or.2016.5033
Pages: 2059-2070

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

20(S)-ginsenoside Rh2 [(S)Rh2] possesses potential to prevent cancer in vitro as well as in vivo, but the underlying mechanism is still unknown. First, we infected HepG2 cells with lentivirus which carries β‑catenin. We detected the pharmacological effects of (S)Rh2 on HepG2 and HepG2‑β‑catenin cells and found that the IC50 of (S)Rh2 exposure on HepG2-β-catenin cells was higher than HepG2 cells. Flow cytometry (FCM) indicated that (S)Rh2 could be arrested in G0/G1 phase and induce early apoptosis in HepG2 and HepG2‑β‑catenin cells. Second, ELISA kit was used to check the activity of glycogen synthase kinase‑3β (GSK‑3β), which was upregulated by (S)Rh2. GSK‑3β inhibitor BIO, was used to verify that (S)Rh2 activated GSK‑3β. PCR and western blotting results indicated that (S)Rh2 could degrade the expression of β‑catenin, which combined with TCF in the nucleus and activate transcription of Wnt target genes, such as Bax, Bcl‑2, cyclin D1, MMP3, which were checked by chromatin immunoprecipitation (ChIP), PCR and western blotting. The results showed that the expression of Bax mRNA and proteins increased, while the cyclin D1, Bcl‑2, MMP3 mRNA and proteins were downregulated in HepG2 and HepG2‑β‑catenin cells which was induced by (S)Rh2. By contrast, with the HepG2-β-catenin + (S)Rh2 group, the expression of other mRNA and proteins in HepG2 + (S)Rh2 group changed significantly. In vivo, experiments were performed using a nude mouse xenograft model to investigate the (S)Rh2 effect. So these results suggested that (S)Rh2 could suppress proliferation, promote apoptosis and inhibit metastasis of HepG2, decrease weight of tumor by downregulating β‑catenin through activating GSK‑3β and the pharmacological effect of (S)Rh2 on HepG2 cells might be weakened by overexpression of β‑catenin.

View Figures

View References

1	Tsai WL, Lai KH, Liang HL, Hsu PI, Chan HH, Chen WC, Yu HC, Tsay FW, Wang HM, Tsai HC, et al: Hepatic arterial infusion chemotherapy for patients with huge unresectable hepatocellular carcinoma. PLoS One. 9:e927842014. View Article : Google Scholar : PubMed/NCBI
2	Page AJ, Cosgrove DC, Philosophe B and Pawlik TM: Hepatocellular carcinoma: Diagnosis, management, and prognosis. Surg Oncol Clin N Am. 23:289–311. 2014. View Article : Google Scholar : PubMed/NCBI
3	Scaggiante B, Kazemi M, Pozzato G, Dapas B, Farra R, Grassi M, Zanconati F and Grassi G: Novel hepatocellular carcinoma molecules with prognostic and therapeutic potentials. World J Gastroenterol. 20:1268–1288. 2014. View Article : Google Scholar : PubMed/NCBI
4	Tanase A-M, Dumitrascu T, Dima S, Grigorie R, Marchio A, Pineau P and Popescu I: Influence of hepatitis viruses on clinicopathological profiles and long-term outcome in patients undergoing surgery for hepatocellular carcinoma. Hepatobiliary Pancreat Dis Int. 13:162–172. 2014. View Article : Google Scholar : PubMed/NCBI
5	Ji Z, Wang T, Shao Z, Huang D, Wang A, Guo Z, Long Y, Zhang L, Su H, Zhang Q, et al: A population-based study examining hepatitis B virus infection and immunization rates in Northwest China. PLoS One. 9:e974742014. View Article : Google Scholar : PubMed/NCBI
6	Xu L-B, Wang J, Liu C, Pang HW, Chen YJ, Ou QJ and Chen JS: Staging systems for predicting survival of patients with hepatocellular carcinoma after surgery. World J Gastroenterol. 16:5257–5262. 2010. View Article : Google Scholar : PubMed/NCBI
7	Osaki A, Suda T, Kamimura K, Tsuchiya A, Tamura Y, Takamura M, Igarashi M, Kawai H, Yamagiwa S and Aoyagi Y: A safe and effective dose of cisplatin in hepatic arterial infusion chemotherapy for hepatocellular carcinoma. Cancer Med. 2:86–98. 2013. View Article : Google Scholar : PubMed/NCBI
8	Ker CG, Chen JS, Kuo KK, Chuang SC, Wang SJ, Chang WC, Lee KT, Chen HY and Juan CC: Liver surgery for hepatocellular carcinoma: Laparoscopic versus open approach. Int J Hepatol. 2011:5967922011. View Article : Google Scholar : PubMed/NCBI
9	Miyahara K, Nouso K and Yamamoto K: Chemotherapy for advanced hepatocellular carcinoma in the sorafenib age. World J Gastroenterol. 20:4151–4159. 2014. View Article : Google Scholar : PubMed/NCBI
10	Song SB, Tung NH, Quang TH, Ngan NT, Kim KE and Kim YH: Inhibition of TNF-α-mediated NF-κB transcriptional activity in HepG2 cells by dammarane-type saponins from Panax ginseng leaves. J Ginseng Res. 36:146–152. 2012. View Article : Google Scholar
11	Guo XX, Guo Q, Li Y, Lee SK, Wei XN and Jin YH: Ginsenoside Rh2 induces human hepatoma cell apoptosisvia bax/bak triggered cytochrome C release and caspase-9/caspase-8 activation. Int J Mol Sci. 13:15523–15535. 2012. View Article : Google Scholar
12	Wang Z, Zheng Q, Liu K, Li G and Zheng R: Ginsenoside Rh(2) enhances antitumour activity and decreases genotoxic effect of cyclophosphamide. Basic Clin Pharmacol Toxicol. 98:411–415. 2006. View Article : Google Scholar : PubMed/NCBI
13	Ho YL, Li KC, Chao W, Chang YS and Huang GJ: Korean red ginseng suppresses metastasis of human hepatoma SK-Hep1 cells by inhibiting matrix metalloproteinase-2/-9 and urokinase plasminogen activator. Evid Based Complement Alternat Med. 2012:9658462012. View Article : Google Scholar : PubMed/NCBI
14	Chung KS, Cho SH, Shin JS, Kim DH, Choi JH, Choi SY, Rhee YK, Hong HD and Lee KT: Ginsenoside Rh2 induces cell cycle arrest and differentiation in human leukemia cells by upregulating TGF-β expression. Carcinogenesis. 34:331–340. 2013. View Article : Google Scholar
15	Yan XJ, Gong LH, Zheng FY, Cheng KJ, Chen ZS and Shi Z: Triterpenoids as reversal agents for anticancer drug resistance treatment. Drug Discov Today. 19:482–488. 2014. View Article : Google Scholar
16	Li S, Gao Y, Ma W, Guo W, Zhou G, Cheng T and Liu Y: EGFR signaling-dependent inhibition of glioblastoma growth by ginsenoside Rh2. Tumour Biol. 35:5593–5598. 2014. View Article : Google Scholar : PubMed/NCBI
17	Park EK, Lee EJ, Lee SH, Koo KH, Sung JY, Hwang EH, Park JH, Kim CW, Jeong KC, Park BK, et al: Induction of apoptosis by the ginsenoside Rh2 by internalization of lipid rafts and caveolae and inactivation of Akt. Br J Pharmacol. 160:1212–1223. 2010. View Article : Google Scholar : PubMed/NCBI
18	Maurer U, Preiss F, Brauns-Schubert P, Schlicher L and Charvet C: GSK-3 - at the crossroads of cell death and survival. J Cell Sci. 127:1369–1378. 2014. View Article : Google Scholar : PubMed/NCBI
19	Zhang JS, Herreros-Villanueva M, Koenig A, Deng Z, de Narvajas AA, Gomez TS, Meng X, Bujanda L, Ellenrieder V, Li XK, et al: Differential activity of GSK-3 isoforms regulates NF-κB and TRAIL- or TNFα induced apoptosis in pancreatic cancer cells. Cell Death Dis. 5:e11422014. View Article : Google Scholar
20	Gao Y, Liu Z, Zhang X, He J, Pan Y, Hao F, Xie L, Li Q, Qiu X and Wang E: Inhibition of cytoplasmic GSK-3β increases cisplatin resistance through activation of Wnt/β-catenin signaling in A549/DDP cells. Cancer Lett. 336:231–239. 2013. View Article : Google Scholar : PubMed/NCBI
21	Lee H, Bae S, Kim YS and Yoon Y: WNT/β-catenin pathway mediates the anti-adipogenic effect of platycodin D, a natural compound found in Platycodon grandiflorum. Life Sci. 89:388–394. 2011. View Article : Google Scholar : PubMed/NCBI
22	Thompson MD and Monga SP: WNT/beta-catenin signaling in liver health and disease. Hepatology. 45:1298–1305. 2007. View Article : Google Scholar : PubMed/NCBI
23	Mosimann C, Hausmann G and Basler K: Beta-catenin hits chromatin: Regulation of Wnt target gene activation. Nat Rev Mol Cell Biol. 10:276–286. 2009. View Article : Google Scholar : PubMed/NCBI
24	Ying Y and Tao Q: Epigenetic disruption of the WNT/beta-catenin signaling pathway in human cancers. Epigenetics. 4:307–312. 2009. View Article : Google Scholar : PubMed/NCBI
25	Chen L, Dai J, Wang Z, Zhang H, Huang Y and Zhao Y: Ginseng total saponins reverse corticosterone-induced changes in depression-like behavior and hippocampal plasticity-related proteins by interfering with GSK-3 β-CREB signaling pathway. Evid Based Complement Alternat Med. 2014:5067352014. View Article : Google Scholar
26	Fu M, Wang C, Rao M, Wu X, Bouras T, Zhang X, Li Z, Jiao X, Yang J, Li A, et al: Cyclin D1 represses p300 transactivation through a cyclin-dependent kinase-independent mechanism. J Biol Chem. 280:29728–29742. 2005. View Article : Google Scholar : PubMed/NCBI
27	Dahmani R, Just PA and Perret C: The Wnt/β-catenin pathway as a therapeutic target in human hepatocellular carcinoma. Clin Res Hepatol Gastroenterol. 35:709–713. 2011. View Article : Google Scholar : PubMed/NCBI
28	Debeb BG, Lacerda L, Xu W, Larson R, Solley T, Atkinson R, Sulman EP, Ueno NT, Krishnamurthy S, Reuben JM, et al: Histone deacetylase inhibitors stimulate dedifferentiation of human breast cancer cells through WNT/β-catenin signaling. Stem Cells. 30:2366–2377. 2012. View Article : Google Scholar : PubMed/NCBI
29	Correia AL, Mori H, Chen EI, Schmitt FC and Bissell MJ: The hemopexin domain of MMP3 is responsible for mammary epithelial invasion and morphogenesis through extracellular interaction with HSP90β. Genes Dev. 27:805–817. 2013. View Article : Google Scholar : PubMed/NCBI
30	Wang X, Li F, Fan C, Wang C and Ruan H: Effects and relationship of ERK1 and ERK2 in interleukin-1β-induced alterations in MMP3, MMP13, type II collagen and aggrecan expression in human chondrocytes. Int J Mol Med. 27:583–589. 2011.PubMed/NCBI
31	Konstantinopoulos PA, Karamouzis MV, Papatsoris AG and Papavassiliou AG: Matrix metalloproteinase inhibitors as anticancer agents. Int J Biochem Cell Biol. 40:1156–1168. 2008. View Article : Google Scholar : PubMed/NCBI