Extracellular signal-regulated kinase and Akt activation play a critical role in the process of hepatocyte growth factor-induced epithelial-mesenchymal transition

Tanahashi,Toshiyuki; Osada,Shinji; Yamada,Atsuko; Kato,Junko; Yawata,Kazunori; Mori,Ryutaro; Imai,Hisashi; Sasaki,Yoshiyuki; Saito,Shiro; Tanaka,Yoshihiro; Nonaka,Kenichi; Yoshida,Kazuhiro

doi:10.3892/ijo.2012.1726

Extracellular signal-regulated kinase and Akt activation play a critical role in the process of hepatocyte growth factor-induced epithelial-mesenchymal transition

Authors:
- Toshiyuki Tanahashi
- Shinji Osada
- Atsuko Yamada
- Junko Kato
- Kazunori Yawata
- Ryutaro Mori
- Hisashi Imai
- Yoshiyuki Sasaki
- Shiro Saito
- Yoshihiro Tanaka
- Kenichi Nonaka
- Kazuhiro Yoshida
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Affiliations: Department of Surgical Oncology, Gifu University Graduate School of Medicine, Gifu, Japan
Published online on: December 3, 2012 https://doi.org/10.3892/ijo.2012.1726
Pages: 556-564

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Abstract

Epithelial-mesenchymal transition (EMT) has recently been studied to elucidate mechanisms of the liver metastatic process. We investigated EMT in the process of liver metastasis and the effects of chemotherapy on EMT cells as therapeutic strategy for colorectal liver metastasis. We used the CT26 murine colorectal carcinoma cell line to create an in vivo mouse liver metastasis model. Liver tumors were stained immunohistochemically. Expression of proteins associated with TGF-β/Smad and hepatocyte growth factor (HGF)/c-Met pathways were investigated by western blotting. Cells with c-Met mRNA knockdown by siRNA techniques showed clearly reduced liver metastases compared with regular cells at 21 days. TGF-β and HGF induced EMT expression, but signal transduction was quite different. TGF-β induced ERK, but not Akt phosphorylation. HGF mediated both ERK and Akt phosphorylation. Akt inhibitor blocked Akt phosphorylation but did not affect TGF-β-induced activation of ERK, Snail and Slug. U-0126 did not reduce Snail activity by TGF-β at a concentration to block ERK phosphorylation. However, Akt inhibitor and U-0126 completely inhibited HGF-induced Slug activation. 5-FU mediated cell death in the EMT process induced by TGF-β more effectively than HGF. ERK/Akt signaling, but not the Smad pathway, may be one of the main processes in HGF-induced EMT, despite the Smad pathway, but not ERK/Akt, being critical for TGF-β-induced EMT. The MAPK/Akt pathway is indispensable in HGF/c-Met signaling. The ERK/Akt pathway particularly may be critical in the HGF-induced EMT process. However, long-term use of chemotherapeutic agents may induce drug resistance and distant metastases through EMT-related signaling pathway activation.

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1.	Bingham S and Riboli E: Diet and cancer - the European prospective investigation into cancer and nutrition. Nat Rev Cancer. 4:206–215. 2004. View Article : Google Scholar : PubMed/NCBI
2.	Benoist S and Nordlinger B: The role of preoperative chemotherapy in patients with resectable colorectal liver metastases. Ann Surg Oncol. 16:2385–2390. 2009. View Article : Google Scholar : PubMed/NCBI
3.	Takeuchi H, Bilchik A, Saha S, et al: c-MET expression level in primary colon cancer: a predictor of tumor invasion and lymph node metastases. Clin Cancer Res. 9:1480–1488. 2003.PubMed/NCBI
4.	Matsui S, Osada S, Tomita H, et al: Clinical significance of aggressive hepatectomy for colorectal liver metastasis, evaluated from the HGF/c-Met pathway. Int J Oncol. 37:289–297. 2010.PubMed/NCBI
5.	Bates RC and Mercurio AM: The epithelial-mesenchymal transition (EMT) and colorectal cancer progression. Cancer Biol Ther. 4:365–370. 2005. View Article : Google Scholar : PubMed/NCBI
6.	Zavadil J and Bottinger EP: TGF-beta and epithelial-to-mesenchymal transitions. Oncogene. 24:5764–5774. 2005. View Article : Google Scholar : PubMed/NCBI
7.	Derynck R and Zhang YE: Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 425:577–584. 2003. View Article : Google Scholar : PubMed/NCBI
8.	Bierie B and Moses HL: Tumour microenvironment: TGFbeta: the molecular Jekyll and Hyde of cancer. Nat Rev Cancer. 6:506–520. 2006. View Article : Google Scholar : PubMed/NCBI
9.	Zhang B, Halder SK, Zhang S and Datta PK: Targeting transforming growth factor-beta signaling in liver metastasis of colon cancer. Cancer Lett. 277:114–120. 2009. View Article : Google Scholar : PubMed/NCBI
10.	Chang HY, Kao MC, Way TD, Ho CT and Fu E: Diosgenin suppresses hepatocyte growth factor (HGF)-induced epithelialmesenchymal transition by down-regulation of Mdm2 and vimentin. J Agric Food Chem. 59:5357–5363. 2011. View Article : Google Scholar : PubMed/NCBI
11.	Osada S, Tomita H, Tanaka Y, et al: The utility of vitamin K3 (menadione) against pancreatic cancer. Anticancer Res. 28:45–50. 2008.PubMed/NCBI
12.	Osada S, Sakashita F, Hosono Y, et al: Extracellular signal-regulated kinase phosphorylation due to menadione-induced arylation mediates growth inhibition of pancreas cancer cells. Cancer Chemother Pharmacol. 62:315–320. 2008. View Article : Google Scholar
13.	Osada S, Saji S and Osada K: Critical role of extracellular signal-regulated kinase phosphorylation on menadione (vitamin K3) induced growth inhibition. Cancer. 91:1156–1165. 2001. View Article : Google Scholar : PubMed/NCBI
14.	Osada S and Carr BI: Mechanism of novel vitamin K analog induced growth inhibition in human hepatoma cell line. J Hepatol. 34:676–682. 2001. View Article : Google Scholar : PubMed/NCBI
15.	Osada S, Osada K and Carr BI: Tumor cell growth inhibition and extracellular signal-regulated kinase (ERK) phosphorylation by novel K vitamins. J Mol Biol. 314:765–772. 2001. View Article : Google Scholar : PubMed/NCBI
16.	Osada S, Sakashita F, Katoh H, Sugiyama Y and Adachi Y: Identification of an immune tolerance reaction in response to pretreatment with frozen pancreatic tissue in islet cell transplantation in rats. Pancreas. 30:e29–e33. 2005. View Article : Google Scholar : PubMed/NCBI
17.	Osada S, Imai H, Tomita H, et al: Vascular endothelial growth factor protects hepatoma cells against oxidative stress-induced cell death. J Gastroenterol Hepatol. 21:988–993. 2006. View Article : Google Scholar : PubMed/NCBI
18.	Thiery JP: Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2:442–454. 2002. View Article : Google Scholar : PubMed/NCBI
19.	Wilmanns C, Steinhauer S, Grossmann J and Ruf G: Site-dependent differences in clinical, pathohistological, and molecular parameters in metastatic colon cancer. Int J Biol Sci. 5:458–465. 2009. View Article : Google Scholar : PubMed/NCBI
20.	Tsuji T, Ibaragi S, Shima K, et al: Epithelial-mesenchymal transition induced by growth suppressor p12CDK2-AP1 promotes tumor cell local invasion but suppresses distant colony growth. Cancer Res. 68:10377–10386. 2008. View Article : Google Scholar
21.	Lee JM, Dedhar S, Kalluri R and Thompson EW: The epithelialmesenchymal transition: new insights in signaling, development, and disease. J Cell Biol. 172:973–981. 2006. View Article : Google Scholar : PubMed/NCBI
22.	Kang Y and Massague J: Epithelial-mesenchymal transitions: twist in development and metastasis. Cell. 118:277–279. 2004. View Article : Google Scholar : PubMed/NCBI
23.	Wu Y and Zhou BP: New insights of epithelial-mesenchymal transition in cancer metastasis. Acta Biochim Biophys Sin. 40:643–650. 2008. View Article : Google Scholar : PubMed/NCBI
24.	Xie LQ, Bian LJ, Li Z, Li Y, Li ZX and Li B: Altered expression of E-cadherin by hepatocyte growth factor and effect on the prognosis of nasopharyngeal carcinoma. Ann Surg Oncol. 17:1927–1936. 2010. View Article : Google Scholar : PubMed/NCBI
25.	Walsh LA and Damjanovski S: IGF-1 increases invasive potential of MCF 7 breast cancer cells and induces activation of latent TGF-beta1 resulting in epithelial to mesenchymal transition. Cell Commun Signal. 9:102011. View Article : Google Scholar : PubMed/NCBI
26.	Ding W, You H, Dang H, et al: Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion. Hepatology. 52:945–953. 2010. View Article : Google Scholar : PubMed/NCBI
27.	Ogunwobi OO and Liu C: Hepatocyte growth factor upregulation promotes carcinogenesis and epithelial-mesenchymal transition in hepatocellular carcinoma via Akt and COX-2 pathways. Clin Exp Metastasis. 28:721–731. 2011. View Article : Google Scholar
28.	Nawshad A, Lagamba D, Polad A and Hay ED: Transforming growth factor-beta signaling during epithelial-mesenchymal transformation: implications for embryogenesis and tumor metastasis. Cells Tissues Organs. 179:11–23. 2005. View Article : Google Scholar
29.	Peinado H, Olmeda D and Cano A: Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer. 7:415–428. 2007. View Article : Google Scholar : PubMed/NCBI
30.	Pon YL, Zhou HY, Cheung AN, Ngan HY and Wong AS: p70 S6 kinase promotes epithelial to mesenchymal transition through snail induction in ovarian cancer cells. Cancer Res. 68:6524–6532. 2008. View Article : Google Scholar : PubMed/NCBI
31.	Miyazono K: Transforming growth factor-β signaling in epithelialmesenchymal transition and progression of cancer. Proc the Japan Academy, Series B. 85:314–323. 2009.
32.	Trusolino L, Bertotti A and Comoglio PM: MET signalling: principles and functions in development, organ regeneration and cancer. Nat Rev Mol Cell Biol. 11:834–848. 2010. View Article : Google Scholar : PubMed/NCBI
33.	Larue L and Bellacosa A: Epithelial-mesenchymal transition in development and cancer: role of phosphatidylinositol 3′ kinase/AKT pathways. Oncogene. 24:7443–7454. 2005.
34.	Vincan E and Barker N: The upstream components of the Wnt signalling pathway in the dynamic EMT and MET associated with colorectal cancer progression. Clin Exp Metastasis. 25:657–663. 2008. View Article : Google Scholar : PubMed/NCBI
35.	Karhadkar SS, Bova GS, Abdallah N, et al: Hedgehog signalling in prostate regeneration, neoplasia and metastasis. Nature. 431:707–712. 2004. View Article : Google Scholar : PubMed/NCBI
36.	Wang Z, Banerjee S, Li Y, Rahman KM, Zhang Y and Sarkar FH: Down-regulation of notch-1 inhibits invasion by inactivation of nuclear factor-kappaB, vascular endothelial growth factor, and matrix metalloproteinase-9 in pancreatic cancer cells. Cancer Res. 66:2778–2784. 2006. View Article : Google Scholar
37.	Fuxe J, Vincent T and Garcia de Herreros A: Transcriptional crosstalk between TGF-beta and stem cell pathways in tumor cell invasion: role of EMT promoting Smad complexes. Cell Cycle. 9:2363–2374. 2010. View Article : Google Scholar : PubMed/NCBI
38.	Creighton CJ, Chang JC and Rosen JM: Epithelial-mesenchymal transition (EMT) in tumor-initiating cells and its clinical implications in breast cancer. J Mammary Gland Biol Neoplasia. 15:253–260. 2010. View Article : Google Scholar : PubMed/NCBI
39.	Mani SA, Guo W, Liao MJ, et al: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 133:704–715. 2008. View Article : Google Scholar : PubMed/NCBI
40.	Singh A and Settleman J: EMT, cancer stem cells and drug resistance: an emerging axis of evil in the war on cancer. Oncogene. 29:4741–4751. 2010. View Article : Google Scholar : PubMed/NCBI
41.	Kajiyama H, Shibata K, Terauchi M, et al: Chemoresistance to paclitaxel induces epithelial-mesenchymal transition and enhances metastatic potential for epithelial ovarian carcinoma cells. Int J Oncol. 31:277–283. 2007.
42.	Yang AD, Fan F, Camp ER, et al: Chronic oxaliplatin resistance induces epithelial-to-mesenchymal transition in colorectal cancer cell lines. Clin Cancer Res. 12:4147–4153. 2006. View Article : Google Scholar : PubMed/NCBI
43.	Tanahashi T, Osada S, Imai H, et al: Signal transduction of vitamin K3 for pancreas cancer therapy. Oncol Rev. 5:57–60. 2010. View Article : Google Scholar
44.	Radisky DC, Levy DD, Littlepage LE, et al: Rac1b and reactive oxygen species mediate MMP-3-induced EMT and genomic instability. Nature. 436:123–127. 2005. View Article : Google Scholar : PubMed/NCBI
45.	Matsubara D, Ishikawa S, Oguni S, Aburatani H, Fukayama M and Niki T: Molecular predictors of sensitivity to the MET inhibitor PHA665752 in lung carcinoma cells. J Thorac Oncol. 5:1317–1324. 2010. View Article : Google Scholar : PubMed/NCBI
46.	Koo JS, Jung W and Jeong J: The predictive role of E-cadherin and androgen receptor on in vitro chemosensitivity in triple-negative breast cancer. Jpn J Clin Oncol. 39:560–568. 2009. View Article : Google Scholar : PubMed/NCBI
47.	Nakamura T, Kato Y, Fuji H, Horiuchi T, Chiba Y and Tanaka K: E-cadherin-dependent intercellular adhesion enhances chemoresistance. Int J Mol Med. 12:693–700. 2003.PubMed/NCBI
48.	Graziano F, Mandolesi A, Ruzzo A, et al: Predictive and prognostic role of E-cadherin protein expression in patients with advanced gastric carcinomas treated with palliative chemotherapy. Tumour Biol. 25:106–110. 2004. View Article : Google Scholar : PubMed/NCBI
49.	Yu SJ, Yu JK, Ge WT, Hu HG, Yuan Y and Zheng S: SPARCL1, Shp2, MSH2, E-cadherin, p53, ADCY-2 and MAPK are prognosis-related in colorectal cancer. World J Gastroenterol. 17:2028–2036. 2011. View Article : Google Scholar : PubMed/NCBI
50.	Osada S, Kanematsu M, Imai H and Goshima S: Clinical significance of serum HGF and c-Met expression in tumor tissue for evaluation of properties and treatment of hepatocellular carcinoma. Hepatogastroenterology. 55:544–549. 2008.PubMed/NCBI