Hypoxia enhances cholangiocarcinoma invasion through activation of hepatocyte growth factor receptor and the extracellular signal‑regulated kinase signaling pathway

Vanichapol,Thitinee; Leelawat,Kawin; Hongeng,Suradej

doi:10.3892/mmr.2015.3865

Hypoxia enhances cholangiocarcinoma invasion through activation of hepatocyte growth factor receptor and the extracellular signal‑regulated kinase signaling pathway

Authors:
- Thitinee Vanichapol
- Kawin Leelawat
- Suradej Hongeng
View Affiliations

Affiliations: Department of Molecular Medicine, Faculty of Science, Mahidol University, Bangkok 10400, Thailand, Department of Surgery, Rajavithi Hospital, Bangkok 10400, Thailand, Department of Pediatrics, Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand
Published online on: May 27, 2015 https://doi.org/10.3892/mmr.2015.3865
Pages: 3265-3272
Copyright: © Vanichapol et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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

Hypoxia is associated with tumor progression and poor prognosis in several cancer types. The present study aimed to examine the contribution of hypoxia (1% O2) to cancer progression in a cholangiocarcinoma cell line, RMCCA‑1. The molecular basis of the hypoxic response pathway was investigated. The results showed that hypoxia significantly accelerated cancer cell proliferation and enhanced cell invasion (P<0.05). By using receptor tyrosine kinase and intracellular signaling antibody array kits, an increased phosphorylation/activation of a number of signaling molecules, particularly hepatocyte growth factor receptor (Met) and extracellular signal‑regulated kinase (ERK) 1/2, was identified. Inhibition of Met and ERK by small hairpin RNA and U0126, respectively, significantly inhibited hypoxia‑induced the invasive potential of RMCCA‑1 cells (P<0.05). However, according to immunohistochemical analysis, hypoxia‑inducible factor‑1α expression was not correlated with cancer staging or tumor differentiation in 44 samples of cholangicarcinoma cases. The findings of the present study emphasized the importance of Met/ERK pathway activation as a key molecular event that may be responsible for a more invasive phenotype in hypoxic tumors and suggest Met as a potential target for the treatment of cholangiocarcinoma.

View Figures

View References

1	Zogragos GN, Farfaras A, Zagouri F, Chrysikos D and Karaliotas K: Cholangiocarcinoma: principles and current trends. Hepatobiliary Pancreat Dis Int. 10:10–20. 2011. View Article : Google Scholar
2	Razumilava N and Gores GJ: Classification, diagnosis and management of cholangiocarcinoma. Clin Gastroenterol Hepatol. 11:13–21. 2013. View Article : Google Scholar
3	Rattanasinganchan P, Leelawat K, Treepongkaruna SA, Tocharoentanaphol C, Subwongcharoen S, Suthiphongchai T and Tohtong R: Establishment and characterization of a cholan-giocarcinoma cell line (RMCCA-1) from a Thai patient. World J Gastroenterol. 12:6500–6506. 2006.PubMed/NCBI
4	Friman S: Cholangiocarcinoma-current treatment options. Scand J Surg. 100:30–34. 2011.
5	Miyoshi A, Kitajima Y, Ide T, et al: Hypoxia accelerates cancer invasion of hepatoma cells by upregulating MMP expression in an HIF-1α-independent manner. Int J Oncol. 29:1533–1539. 2006.PubMed/NCBI
6	Yokoi K and Fidler IJ: Hypoxia increases resistance of human pancreatic cancer cells to apoptosis induced by gemcitabine. Clin Cancer Res. 10:2299–2306. 2004. View Article : Google Scholar : PubMed/NCBI
7	Kim JY, Kim YJ, Lee S and Park JH: The critical role of ERK in death resistance and invasiveness of hypoxia-selected glioblastoma cells. BMC Cancer. 9:272009. View Article : Google Scholar : PubMed/NCBI
8	Kitajima Y, Ide T, Ohtsuka T and Miyazaki K: Induction of hepatocyte growth factor activator gene expression under hypoxia activates the hepatocyte growth factor/c-Met system via hypoxia inducible factor-1 in pancreatic cancer. Cancer Sci. 99:1341–1347. 2008. View Article : Google Scholar : PubMed/NCBI
9	Koh MY, Lemos R Jr, Lui X and Powis G: The hypoxia-associated factor switches cells from HIF-1α- to HIF-2α-dependent signaling promoting stem cell characteristics, aggressive tumor growth and invasion. Cancer Res. 71:4015–4027. 2011. View Article : Google Scholar : PubMed/NCBI
10	Fu OY, Hou MF, Yang SF, Huang SC and Lee WY: Cobalt chloride-induced hypoxia modulates the invasive potential and matrix matelloproteinases of primary and metastatic breast cancer cells. Anticancer Res. 29:3131–3138. 2009.PubMed/NCBI
11	Pennacchietti S, Michieli P, Galluzzo M, Mazzone M, Giordano S and Comoglio PM: Hypoxia promotes invasive growth by transcriptional activation of the met protooncogene. Cancer Cell. 3:347–361. 2003. View Article : Google Scholar : PubMed/NCBI
12	Cecchi F, Rabe DC and Bottaro DP: Targeting the HGF/Met signaling pathway in cancer. Eur J Cancer. 46:1260–1270. 2010. View Article : Google Scholar : PubMed/NCBI
13	Socoteanu MP, Mott F, Alpini G and Frankel AE: c-Met targeted therapy of cholangiocarcinoma. World J Gastroenterol. 14:2990–2994. 2008. View Article : Google Scholar : PubMed/NCBI
14	Matsumura A, Kubota T, Taiyoh H, et al: HGF regulates VEGF expression via the c-Met receptor downstream pathways, PI3K/Akt, MAPK and STAT3, in CT26 murine cells. Int J Oncol. 42:535–542. 2013.
15	Leelawat K, Leelawat S, Tepaksorn P, Rattanasinganchan P, Leungchaweng A, Tohtong R and Sobhon P: Involvement of c-Met/hepatocyte growth factor pathway in cholangiocarcinoma cell invasion and its therapeutic inhibition with small interfering RNA specific for c-Met. J Surg Res. 136:78–84. 2006. View Article : Google Scholar : PubMed/NCBI
16	Santarpia L, Lippman SL and El-Naggar AK: Targeting the MAPK-RAS-RAF signaling pathway in cancer therapy. Expert Opin Ther Targets. 16:103–119. 2012. View Article : Google Scholar : PubMed/NCBI
17	Katz M, Amit I and Yarden Y: Regulation of MAPKs by growth factors and receptor tyrosine kinases. Biochim Biophys Acta. 1773:1161–1176. 2007. View Article : Google Scholar : PubMed/NCBI
18	Lee JW, Bae SH, Jeong JW, Kim SH and Kim KW: Hypoxia-inducible factor (HIF-1)α: its protein stability and biological functions. Exp Mol Med. 36:1–12. 2004. View Article : Google Scholar : PubMed/NCBI
19	Harris AL: Hypoxia-a key regulatory factor in tumour growth. Nat Rev Cancer. 2:38–47. 2002. View Article : Google Scholar : PubMed/NCBI
20	Organ SL and Tsao MS: An overview of the Met signaling pathway. Ther Adv Med Oncol. 3(Suppl 1): S7–S19. 2011. View Article : Google Scholar
21	Seubwai W, Kraiklang R, Wongkham C and Wongkham S: Hypoxia enhances aggressiveness of cholangiocarcinoma cells. Asian Pac J Cancer Prev. 13(Suppl): 53–58. 2012.PubMed/NCBI
22	Gwak GY, Yoon JH, Kim KM, Lee HS, Chung JW and Gores GJ: Hypoxia stimulates proliferation of human hepatoma cells through the induction of hexokinase II expression. J Hepatol. 42:358–364. 2005. View Article : Google Scholar : PubMed/NCBI
23	He G, Jiang Y, Zhang B and Wu G: The effect of HIF-1α on glucose metabolism, growth and apoptosis of pancreatic cancerous cells. Asia Pac J Clin Nutr. 23:174–180. 2014.
24	Dai R, Li J, Fu J, et al: The tyrosine kinase c-Met contributes to the pro-tumorigenic function of the p38 kinase in human bile duct cholangiocarcinoma cells. J Biol Chem. 287:39812–39823. 2012. View Article : Google Scholar : PubMed/NCBI
25	Xu L, Nilsson MB, Saintigny P, et al: Epidermal growth factor receptor regulates MET levels and invasiveness through hypoxia-inducible factor-1α in non-small cell lung cancer cells. Oncogene. 29:2616–2627. 2010. View Article : Google Scholar : PubMed/NCBI
26	Mylonis I, Chachami G, Samiotaka M, et al: Identification of MAPK phosphorylation sites and their role in the localization and activity of hypoxia-inducible factor-1α. J Biol Chem. 281:33095–33106. 2006. View Article : Google Scholar : PubMed/NCBI
27	Lee KH, Choi EY, Hyun MS and Kim JR: Involvement of MAPK pathway in hypoxia-induced up-regulation of urokinase plasminogen activator receptor in a human prostatic cancer cell line, PC3MLN4. Exp Mol Med. 36:57–64. 2004. View Article : Google Scholar : PubMed/NCBI
28	Mazzone M and Comoglio PM: The Met pathway: master switch and drug target in cancer progression. FASEB J. 20:1611–1621. 2006. View Article : Google Scholar : PubMed/NCBI
29	Eckerich C, Zapf S, Fillbrandt R, Loges S, Westphat M and Lamszus K: Hypoxia can induce c-Met expression in glioma cells and enhance SF/HGF-induced cell migration. Int J Cancer. 121:276–283. 2007. View Article : Google Scholar : PubMed/NCBI
30	Sullivan R and Graham CH: Hypoxia-driven selection of the metastatic phenotype. Cancer Metastasis Rev. 26:319–331. 2007. View Article : Google Scholar : PubMed/NCBI
31	Hara S, Nakashiro K, Klosek SK, Ishikawa T, Shintani S and Hamakawa H: Hypoxia enhances c-Met/HGF receptor expression and signaling by activating HIF-1alpha in human salivary gland cancer cells. Oral Oncol. 42:593–598. 2006. View Article : Google Scholar : PubMed/NCBI
32	Wang Y, Li Z, Zhang H, et al: HIF-1α and HIF-2α correlate with migration and invasion in gastric cancer. Cancer Biol Ther. 10:376–382. 2010. View Article : Google Scholar : PubMed/NCBI