Purified vitexin compound 1 inhibits growth and angiogenesis through activation of FOXO3a by inactivation of Akt in hepatocellular carcinoma

Wang,Jiangang; Zheng,Xingxing; Zeng,Guangyao; Zhou,Yingjun; Yuan,Hong

doi:10.3892/ijmm.2013.1587

Purified vitexin compound 1 inhibits growth and angiogenesis through activation of FOXO3a by inactivation of Akt in hepatocellular carcinoma

Authors:
- Jiangang Wang
- Xingxing Zheng
- Guangyao Zeng
- Yingjun Zhou
- Hong Yuan
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Affiliations: Department of Cardiology, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China, School of Pharmaceutical Science, Central South University, Changsha, Hunan 410013, P.R. China
Published online on: December 12, 2013 https://doi.org/10.3892/ijmm.2013.1587
Pages: 441-448

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Abstract

Vitexins, isolated from the seeds of the Chinese herb Vitex negundo, is known to exert antitumor activity in cancer xenograft models and cell lines. The aim of the current study was to examine whether the Akt/forkhead box protein O3a (FOXO3a) pathway mediates the biological effects of purified vitexin compound 1 (VB-1) in hepatocellular carcinoma (HCC) cells. The effect of VB-1 on the viability of the HCC cell lines HepG2, Hep3B, Huh-7 and the human embryonic liver cells L-02 was investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Growth inhibition was assessed by clonogenic assay, and cell cycle arrest was investigated using flow cytometry. Inhibition of angiogenesis was evaluated using a matrigel in vitro HUVEC tube formation assay. The effects on the Akt/FOXO3a pathway were detected by western blotting. VB-1 suppressed the proliferation of HepG2, Hep3B, Huh-7 cells, but had little effect on L-02 cells. VB-1 inhibited anchorage-dependent and -independent HepG2 cell growth in a concentration-dependent manner by induction of cell cycle arrest at G1/G0. VB-1 also reduced the secretion of vascular endothelial growth factor (VEGF), resulting in the inhibition of endothelial tube formation. Phosphorylated Akt and its downstream effector FOXO3a were downregulated in VB-1-treated HepG2 cells. Knockdown of Akt1 by small interfering RNA (siRNA) enhanced growth inhibition, and silencing FOXO3a by siRNA attenuated this action. VB-1 inhibited growth and induced cell cycle arrest at G1/G0 by regulating the Akt/FOXO3a pathway. The findings suggested that VB-1 is a potentially promising candidate for the treatment of HCC.

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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
2	Okuda K: Hepatocellular carcinoma. J Hepatol. 32:225–237. 2000. View Article : Google Scholar
3	Llovet JM, Burroughs A and Bruix J: Hepatocellular carcinoma. Lancet. 362:1907–1917. 2003. View Article : Google Scholar
4	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
5	Lai EC and Lau WY: The continuing challenge of hepatic cancer in Asia. Surgeon. 3:210–215. 2005. View Article : Google Scholar : PubMed/NCBI
6	Thompson LU, Chen JM, Li T, Strasser-Weippl K and Goss PE: Dietary flaxseed alters tumor biological markers in postmenopausal breast cancer. Clin Cancer Res. 11:3828–3835. 2005. View Article : Google Scholar : PubMed/NCBI
7	Park JB, Lee MS, Cha EY, et al: Magnolol-induced apoptosis in HCT-116 colon cancer cells is associated with the AMP-activated protein kinase signaling pathway. Biol Pharm Bull. 35:1614–1620. 2012.PubMed/NCBI
8	Kuijsten A, Arts IC, Hollman PC, van’t Veer P and Kampman E: Plasma enterolignans are associated with lower colorectal adenoma risk. Cancer Epidemiol Biomarkers Prev. 15:1132–1136. 2006. View Article : Google Scholar : PubMed/NCBI
9	Adlercreutz H: Lignans and human health. Crit Rev Clin Lab Sci. 44:483–525. 2007. View Article : Google Scholar
10	Bergman Jungestrom M, Thompson LU and Dabrosin C: Flaxseed and its lignans inhibit estradiol-induced growth, angiogenesis, and secretion of vascular endothelial growth factor in human breast cancer xenografts in vivo. Clin Cancer Res. 13:1061–1067. 2007.PubMed/NCBI
11	Cuca LE, Coy ED, Alarcón MA, Fernández A and Aristizábal FA: Cytotoxic effect of some natural compounds isolated from Lauraceae plants and synthetic derivatives. Biomedica. 31:335–343. 2011.PubMed/NCBI
12	Zhou Y, Liu YE, Cao J, et al: Vitexins, nature-derived lignan compounds, induce apoptosis and suppress tumor growth. Clin Cancer Res. 15:5161–5169. 2009. View Article : Google Scholar : PubMed/NCBI
13	Liu Z, Xiao Y, Yuan Y, et al: Effects of oleic acid on cell proliferation through an integrin-linked kinase signaling pathway in 786-O renal cell carcinoma cells. Oncol Lett. 5:1395–1399. 2013.PubMed/NCBI
14	Vene R, Benelli R, Minghelli S, Astigiano S, Tosetti F and Ferrari N: Xanthohumol impairs human prostate cancer cell growth and invasion and diminishes the incidence and progression of advanced tumors in TRAMP mice. Mol Med. 18:1292–1302. 2012. View Article : Google Scholar : PubMed/NCBI
15	Rubinfeld H, Cohen-Kaplan V, Nass D, et al: Heparanase is highly expressed and regulates proliferation in GH-secreting pituitary tumor cells. Endocrinology. 152:4562–4570. 2011. View Article : Google Scholar : PubMed/NCBI
16	Wu J, Wu X, Zhong D, Zhai W, Ding Z and Zhou Y: Short hairpin RNA (shRNA) Ether à go-go 1 (Eag1) inhibition of human osteosarcoma angiogenesis via VEGF/PI3K/AKT signaling. Int J Mol Sci. 13:12573–12583. 2012.
17	Khaidakov M and Mehta JL: Oxidized LDL triggers pro-oncogenic signaling in human breast mammary epithelial cells partly via stimulation of MiR-21. PLoS One. 7:e469732012. View Article : Google Scholar : PubMed/NCBI
18	van Dijk M, van Bezu J, van Abel D, et al: The STOX1 genotype associated with pre-eclampsia leads to a reduction of trophoblast invasion by alpha-T-catenin upregulation. Hum Mol Genet. 19:2658–2667. 2010.PubMed/NCBI
19	Biggs WH III, Meisenhelder J, Hunter T, Cavenee WK and Arden KC: Protein kinase B/Akt-mediated phosphorylation promotes nuclear exclusion of the winged helix transcription factor FKHR1. Proc Natl Acad Sci USA. 96:7421–7426. 1999. View Article : Google Scholar
20	Band AM, Björklund M and Laiho M: The phosphatidylinositol 3-kinase/Akt pathway regulates transforming growth factor-{beta} signaling by destabilizing ski and inducing Smad7. J Biol Chem. 284:35441–35449. 2009.
21	Wang L, Jia D, Duan F, et al: Combined anti-tumor effects of IFN-alpha and sorafenib on hepatocellular carcinoma in vitro and in vivo. Biochem Biophys Res Commun. 422:687–692. 2012. View Article : Google Scholar : PubMed/NCBI
22	Vanbrocklin MW, Robinson JP, Lastwika KJ, McKinney AJ, Gach HM and Holmen SL: Ink4a/Arf loss promotes tumor recurrence following Ras inhibition. Neuro Oncol. 14:34–42. 2012. View Article : Google Scholar : PubMed/NCBI
23	Kim BC: FoxO3a mediates transforming growth factor-beta1-induced apoptosis in FaO rat hepatoma cells. BMB Rep. 41:728–732. 2008. View Article : Google Scholar : PubMed/NCBI
24	Wang S, Chong ZZ, Shang YC and Maiese K: WISP1 neuroprotection requires FoxO3a post-translational modulation with autoregulatory control of SIRT1. Curr Neurovasc Res. 10:54–69. 2013. View Article : Google Scholar : PubMed/NCBI
25	Liang C, Chen W, Zhi X, et al: Serotonin promotes the proliferation of serum-deprived hepatocellular carcinoma cells via upregulation of FOXO3a. Mol Cancer. 12:142013. View Article : Google Scholar : PubMed/NCBI
26	Naka K, Hoshii T, Muraguchi T, et al: TGF-beta-FOXO signalling maintains leukaemia-initiating cells in chronic myeloid leukaemia. Nature. 463:676–680. 2010. View Article : Google Scholar : PubMed/NCBI
27	Yang Y, Jiang H, Gao H, et al: The monoclonal antibody CH12 enhances the sorafenib-mediated growth inhibition of hepatocellular carcinoma xenografts expressing epidermal growth factor receptor variant III. Neoplasia. 14:509–518. 2012.
28	Chung YW, Kim HK, Kim IY, Yim MB and Chock PB: Dual function of protein kinase C (PKC) in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced manganese superoxide dismutase (MnSOD) expression: activation of CREB and FOXO3a by PKC-alpha phosphorylation and by PKC-mediated inactivation of Akt, respectively. J Biol Chem. 286:29681–29690. 2011.
29	Kashiwagi A, Fein MJ and Shimada M: Calpain modulates cyclin-dependent kinase inhibitor 1B (p27(Kip1)) in cells of the osteoblast lineage. Calcif Tissue Int. 89:36–42. 2011. View Article : Google Scholar : PubMed/NCBI
30	Dormond O, Madsen JC and Briscoe DM: The effects of mTOR-Akt interactions on anti-apoptotic signaling in vascular endothelial cells. J Biol Chem. 282:23679–23686. 2007. View Article : Google Scholar : PubMed/NCBI
31	Davis R, Singh KP, Kurzrock R and Shankar S: Sulforaphane inhibits angiogenesis through activation of FOXO transcription factors. Oncol Rep. 22:1473–1478. 2009.PubMed/NCBI
32	Wu HH, Wu JY, Cheng YW, et al: cIAP2 upregulated by E6 oncoprotein via epidermal growth factor receptor/phosphatidylinositol 3-kinase/AKT pathway confers resistance to cisplatin in human papillomavirus 16/18-infected lung cancer. Clin Cancer Res. 16:5200–5210. 2010. View Article : Google Scholar : PubMed/NCBI
33	Kreutzer JN, Ruzzene M and Guerra B: Enhancing chemosensitivity to gemcitabine via RNA interference targeting the catalytic subunits of protein kinase CK2 in human pancreatic cancer cells. BMC Cancer. 10:4402010. View Article : Google Scholar : PubMed/NCBI
34	Fleischer B, Schulze-Bergkamen H, Schuchmann M, et al: Mcl-1 is an anti-apoptotic factor for human hepatocellular carcinoma. Int J Oncol. 28:25–32. 2006.PubMed/NCBI
35	He L, Yang X, Cao X, Liu F, Quan M and Cao J: Casticin induces growth suppression and cell cycle arrest through activation of FOXO3a in hepatocellular carcinoma. Oncol Rep. 29:103–108. 2013.PubMed/NCBI
36	Nayak G and Cooper GM: p53 is a major component of the transcriptional and apoptotic program regulated by PI 3-kinase/Akt/GSK3 signaling. Cell Death Dis. 3:e4002012. View Article : Google Scholar : PubMed/NCBI