The inhibitory effect of anthocyanins on Akt on invasion and epithelial-mesenchymal transition is not associated with the anti-EGFR effect of the anthocyanins

Lu,Jing Nan; Lee,Won Sup; Kim,Min Jeong; Yun,Jeong Won; Jung,Ji Hyun; Yi,Sang Mi; Jeong,Jae-Hoon; Kim,Hye Jung; Choi,Yung Hyun; Kim,Gon‑Sup; Ryu,Chung Ho; Shin,Sung Chul

doi:10.3892/ijo.2014.2315

The inhibitory effect of anthocyanins on Akt on invasion and epithelial-mesenchymal transition is not associated with the anti-EGFR effect of the anthocyanins

Authors:
- Jing Nan Lu
- Won Sup Lee
- Min Jeong Kim
- Jeong Won Yun
- Ji Hyun Jung
- Sang Mi Yi
- Jae-Hoon Jeong
- Hye Jung Kim
- Yung Hyun Choi
- Gon‑Sup Kim
- Chung Ho Ryu
- Sung Chul Shin
View Affiliations

Affiliations: Department of Internal Medicine, Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju, Republic of Korea , Research Center for Radiotherapy, Korea Institute of Radiological and Medical Sciences, Seoul, Republic of Korea, Department of Pharmacology, Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju, Republic of Korea , Department of Biochemistry, Dongeui University College of Oriental Medicine and Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan, Republic of Korea, School of Veterinary Medicine, Gyeongsang National University, Jinju, Republic of Korea, Division of Applied Life Science (BK 21 Program), Institute of Agriculture and Life Science, Gyeongsang National University, Jinju, Republic of Korea, Department of Chemistry and Research Institute of Life Science, Gyeongsang National University, Jinju, Republic of Korea
Published online on: February 27, 2014 https://doi.org/10.3892/ijo.2014.2315
Pages: 1756-1766

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

Evidence suggests that anthocyanins inhibit EGFR and Akt activity. However, it is still unknown whether the inhibitory effect of anthocyanins on Akt is associated with the anti-EGFR effect. The effect of anthocyanins on epithelial-mesenchymal transition (EMT) has not been extensively studied. Therefore, we investigated the effects of anthocyanins from fruits of Vitis coignetiae Pulliat (AIMs) on EGF-induced EMT and the underlying molecular mechanisms. AIMs suppressed the invasion of A549 cells in a dose-dependent manner. AIMs inhibited the phosphorylation of Akt and EGFR, but the inhibitory effect on Akt was not derived from EGFR. EGF re-induced Akt phosphorylation at Thr308 in the AIM-treated cells, but not Akt phosphorylation at Ser473. AIMs also inhibited EMT of cancer cells. AIMs inhibited glycogen synthase kinase-3β phosphorylation and β-catenin expression that are invovled in EMT. We confirmed these findings with transforming growth factor (TGF)-β. In conclusion, these data suggest that the inhibitory effect of AIMs on Akt activity is independent of EGFR, and that AIMs suppressed invasion and migration at least in part by suppressing EMT by inhibiting Akt activity as well as EGFR. This study provides evidence that AIMs may have anticancer effects on human cancer cells.

View Figures

View References

1.	Wells A: Tumor invasion: role of growth factor-induced cell motility. Adv Cancer Res. 78:31–101. 2000. View Article : Google Scholar : PubMed/NCBI
2.	Price JT, Wilson HM and Haites NE: Epidermal growth factor (EGF) increases the in vitro invasion, motility and adhesion interactions of the primary renal carcinoma cell line, A704. Eur J Cancer. 32A:1977–1982. 1996. View Article : Google Scholar : PubMed/NCBI
3.	Sieg DJ, Hauck CR, Ilic D, Klingbeil CK, Schaefer E, Damsky CH, et al: FAK integrates growth-factor and integrin signals to promote cell migration. Nat Cell Biol. 2:249–256. 2000. View Article : Google Scholar : PubMed/NCBI
4.	Gullick WJ: Prevalence of aberrant expression of the epidermal growth factor receptor in human cancers. Br Med Bull. 47:87–98. 1991.PubMed/NCBI
5.	Salomon DS, Brandt R, Ciardiello F and Normanno N: Epidermal growth factor-related peptides and their receptors in human malignancies. Crit Rev Oncol Hematol. 19:183–232. 1995. View Article : Google Scholar : PubMed/NCBI
6.	Rhim AD, Mirek ET, Aiello NM, Maitra A, Bailey JM, McAllister F, et al: EMT and dissemination precede pancreatic tumor formation. Cell. 148:349–361. 2012. View Article : Google Scholar : PubMed/NCBI
7.	Meiers S, Kemeny M, Weyand U, Gastpar R, von Angerer E and Marko D: The anthocyanidins cyanidin and delphinidin are potent inhibitors of the epidermal growth-factor receptor. J Agric Food Chem. 49:958–962. 2001. View Article : Google Scholar : PubMed/NCBI
8.	Shin DY, Lee WS, Lu JN, Kang MH, Ryu CH, Kim GY, et al: Induction of apoptosis in human colon cancer HCT-116 cells by anthocyanins through suppression of Akt and activation of p38-MAPK. Int J Oncol. 35:1499–1504. 2009.PubMed/NCBI
9.	Yun JW, Lee WS, Kim MJ, Lu JN, Kang MH, Kim HG, et al: Characterization of a profile of the anthocyanins isolated from Vitis coignetiae Pulliat and their anti-invasive activity on HT-29 human colon cancer cells. Food Chem Toxicol. 48:903–909. 2010.PubMed/NCBI
10.	Vihinen P and Kahari VM: Matrix metalloproteinases in cancer: prognostic markers and therapeutic targets. Int J Cancer. 99:157–166. 2002. View Article : Google Scholar : PubMed/NCBI
11.	Deryugina EI and Quigley JP: Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev. 25:9–34. 2006. View Article : Google Scholar
12.	Goldkorn T, Balaban N, Matsukuma K, Chea V, Gould R, Last J, et al: EGF-receptor phosphorylation and signaling are targeted by H₂O₂ redox stress. Am J Respir Cell Mol Biol. 19:786–798. 1998. View Article : Google Scholar : PubMed/NCBI
13.	Carpenter G and Cohen S: Epidermal growth factor. J Biol Chem. 265:7709–7712. 1990.
14.	Margolis B, Bellot F, Honegger AM, Ullrich A, Schlessinger J and Zilberstein A: Tyrosine kinase activity is essential for the association of phospholipase C-gamma with the epidermal growth factor receptor. Mol Cell Biol. 10:435–441. 1990.PubMed/NCBI
15.	Li Y, Welm B, Podsypanina K, Huang S, Chamorro M, Zhang X, et al: Evidence that transgenes encoding components of the Wnt signaling pathway preferentially induce mammary cancers from progenitor cells. Proc Natl Acad Sci USA. 100:15853–15858. 2003. View Article : Google Scholar
16.	Chu EY, Hens J, Andl T, Kairo A, Yamaguchi TP, Brisken C, et al: Canonical WNT signaling promotes mammary placode development and is essential for initiation of mammary gland morphogenesis. Development. 131:4819–4829. 2004. View Article : Google Scholar : PubMed/NCBI
17.	Jope RS, Yuskaitis CJ and Beurel E: Glycogen synthase kinase-3 (GSK3): inflammation, diseases, and therapeutics. Neurochem Res. 32:577–595. 2007. View Article : Google Scholar : PubMed/NCBI
18.	Teller N, Thiele W, Marczylo TH, Gescher AJ, Boettler U, Sleeman J, et al: Suppression of the kinase activity of receptor tyrosine kinases by anthocyanin-rich mixtures extracted from bilberries and grapes. J Agric Food Chem. 57:3094–3101. 2009. View Article : Google Scholar : PubMed/NCBI
19.	Huang HP, Shih YW, Chang YC, Hung CN and Wang CJ: Chemoinhibitory effect of mulberry anthocyanins on melanoma metastasis involved in the Ras/PI3K pathway. J Agric Food Chem. 56:9286–9293. 2008. View Article : Google Scholar : PubMed/NCBI
20.	Nurwidya F, Takahashi F, Murakami A and Takahashi K: Epithelial mesenchymal transition in drug resistance and metastasis of lung cancer. Cancer Res Treat. 44:151–156. 2012. View Article : Google Scholar : PubMed/NCBI
21.	Barrallo-Gimeno A and Nieto MA: The Snail genes as inducers of cell movement and survival: implications in development and cancer. Development. 132:3151–3161. 2005. View Article : Google Scholar
22.	Kim D, Kim S, Koh H, Yoon SO, Chung AS, Cho KS, et al: Akt/PKB promotes cancer cell invasion via increased motility and metalloproteinase production. FASEB J. 15:1953–1962. 2001. View Article : Google Scholar : PubMed/NCBI
23.	Peinado H, Quintanilla M and Cano A: Transforming growth factor beta-1 induces snail transcription factor in epithelial cell lines: mechanisms for epithelial mesenchymal transitions. J Biol Chem. 278:21113–21123. 2003. View Article : Google Scholar : PubMed/NCBI
24.	Lee J, Moon HJ, Lee JM and Joo CK: Smad3 regulates Rho signaling via NET1 in the transforming growth factor-beta-induced epithelial-mesenchymal transition of human retinal pigment epithelial cells. J Biol Chem. 285:26618–26627. 2010. View Article : Google Scholar
25.	Willis BC and Borok Z: TGF-beta-induced EMT: mechanisms and implications for fibrotic lung disease. Am J Physiol Lung Cell Mol Physiol. 293:L525–L534. 2007. View Article : Google Scholar : PubMed/NCBI
26.	Qiu Q, Yang M, Tsang BK and Gruslin A: EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways. Reproduction. 128:355–363. 2004. View Article : Google Scholar : PubMed/NCBI
27.	Bouchard F, Belanger SD, Biron-Pain K and St-Pierre Y: EGR-1 activation by EGF inhibits MMP-9 expression and lymphoma growth. Blood. 116:759–766. 2010. View Article : Google Scholar : PubMed/NCBI
28.	Stuelten CH, DaCosta Byfield S, Arany PR, Karpova TS, Stetler-Stevenson WG and Roberts AB: Breast cancer cells induce stromal fibroblasts to express MMP-9 via secretion of TNF-alpha and TGF-beta. J Cell Sci. 118:2143–2153. 2005. View Article : Google Scholar : PubMed/NCBI
29.	Wang H, Wang HS, Zhou BH, Li CL, Zhang F, Wang XF, et al: Epithelial-mesenchymal transition (EMT) induced by TNF-alpha requires Akt/GSK-3beta-mediated stabilization of snail in colorectal cancer. PLoS One. 8:e566642013. View Article : Google Scholar : PubMed/NCBI
30.	Yan D, Avtanski D, Saxena NK and Sharma D: Leptin-induced epithelial-mesenchymal transition in breast cancer cells requires beta-catenin activation via Akt/GSK3- and MTA1/Wnt1 protein-dependent pathways. J Biol Chem. 287:8598–8612. 2012. View Article : Google Scholar
31.	Saito Y, Vandenheede JR and Cohen P: The mechanism by which epidermal growth factor inhibits glycogen synthase kinase 3 in A431 cells. Biochem J. 303:27–31. 1994.PubMed/NCBI
32.	Chen G, Bower KA, Xu M, Ding M, Shi X, Ke ZJ, et al: Cyanidin-3-glucoside reverses ethanol-induced inhibition of neurite outgrowth: role of glycogen synthase kinase 3 beta. Neurotox Res. 15:321–331. 2009. View Article : Google Scholar : PubMed/NCBI
33.	Hudson TS, Hartle DK, Hursting SD, Nunez NP, Wang TT, Young HA, et al: Inhibition of prostate cancer growth by muscadine grape skin extract and resveratrol through distinct mechanisms. Cancer Res. 67:8396–8405. 2007. View Article : Google Scholar : PubMed/NCBI
34.	Galbaugh T, Cerrito MG, Jose CC and Cutler ML: EGF-induced activation of Akt results in mTOR-dependent p70S6 kinase phosphorylation and inhibition of HC11 cell lactogenic differentiation. BMC Cell Biol. 7:342006. View Article : Google Scholar : PubMed/NCBI