Taurolithocholic acid promotes intrahepatic cholangiocarcinoma cell growth via muscarinic acetylcholine receptor and EGFR/ERK1/2 signaling pathway

Amonyingcharoen,Sumet; Suriyo,Tawit; Thiantanawat,Apinya; Watcharasit,Piyajit; Satayavivad,Jutamaad

doi:10.3892/ijo.2015.2939

Taurolithocholic acid promotes intrahepatic cholangiocarcinoma cell growth via muscarinic acetylcholine receptor and EGFR/ERK1/2 signaling pathway

Authors:
- Sumet Amonyingcharoen
- Tawit Suriyo
- Apinya Thiantanawat
- Piyajit Watcharasit
- Jutamaad Satayavivad
View Affiliations

Affiliations: Chulabhorn Graduate Institute, Bangkok 10210, Thailand, Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok 10210, Thailand
Published online on: March 27, 2015 https://doi.org/10.3892/ijo.2015.2939
Pages: 2317-2326
Copyright: © Amonyingcharoen 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

Cholangiocarcinoma (CCA) is a malignant cancer of the biliary tract and its occurrence is associated with chronic cholestasis which causes an elevation of bile acids in the liver and bile duct. The present study aimed to investigate the role and mechanistic effect of bile acids on the CCA cell growth. Intrahepatic CCA cell lines, RMCCA-1 and HuCCA-1, were treated with bile acids and their metabolites to determine the growth promoting effect. Cell viability, cell cycle analysis, EdU incorporation assays were conducted. Intracellular signaling proteins were detected by western immunoblotting. Among eleven forms of bile acids and their metabolites, only taurolithocholic acid (TLCA) concentration dependently (1-40 µM) increased the cell viability of RMCCA-1, but not HuCCA-1 cells. The cell cycle analysis showed induction of cells in the S phase and the EdU incorporation assay revealed induction of DNA synthesis in the TLCA-treated RMCCA-1 cells. Moreover, TLCA increased the phosphorylation of EGFR, ERK 1/2 and also increased the expression of cyclin D1 in RMCCA-1 cells. Furthermore, TLCA-induced RMCCA-1 cell growth could be inhibited by atropine, a non-selective muscarinic acetylcholine receptor (mAChR) antagonist, AG 1478, a specific EGFR inhibitor, or U 0126, a specific MEK 1/2 inhibitor. These results suggest that TLCA induces CCA cell growth via mAChR and EGFR/EKR1/2 signaling pathway. Moreover, the functional presence of cholinergic system plays a certain role in TLCA-induced CCA cell growth.

View Figures

View References

1	Patel T: Cholangiocarcinoma - controversies and challenges. Nat Rev Gastroenterol Hepatol. 8:189–200. 2011. View Article : Google Scholar : PubMed/NCBI
2	Raufman JP, Cheng K and Zimniak P: Activation of muscarinic receptor signaling by bile acids: Physiological and medical implications. Dig Dis Sci. 48:1431–1444. 2003. View Article : Google Scholar : PubMed/NCBI
3	Bernstein H, Bernstein C, Payne CM, Dvorakova K and Garewal H: Bile acids as carcinogens in human gastrointestinal cancers. Mutat Res. 589:47–65. 2005. View Article : Google Scholar : PubMed/NCBI
4	Cheng K and Raufman JP: Bile acid-induced proliferation of a human colon cancer cell line is mediated by transactivation of epidermal growth factor receptors. Biochem Pharmacol. 70:1035–1047. 2005. View Article : Google Scholar : PubMed/NCBI
5	Fujino T, Takeuchi A, Maruko-Ohtake A, Ohtake Y, Satoh J, Kobayashi T, Tanaka T, Ito H, Sakamaki R, Kashimura R, et al: Critical role of farnesoid X receptor for hepatocellular carcinoma cell proliferation. J Biochem. 152:577–586. 2012. View Article : Google Scholar : PubMed/NCBI
6	Liu R, Zhao R, Zhou X, Liang X, Campbell DJ, Zhang X, Zhang L, Shi R, Wang G, Pandak WM, et al: Conjugated bile acids promote cholangiocarcinoma cell invasive growth via activation of sphingosine 1-phosphate receptor 2. Hepatology. 60:908–918. 2014. View Article : Google Scholar : PubMed/NCBI
7	Casaburi I, Avena P, Lanzino M, Sisci D, Giordano F, Maris P, Catalano S, Morelli C and Andò S: Chenodeoxycholic acid through a TGR5-dependent CREB signaling activation enhances cyclin D1 expression and promotes human endometrial cancer cell proliferation. Cell Cycle. 11:2699–2710. 2012. View Article : Google Scholar : PubMed/NCBI
8	Cheng K, Chen Y, Zimniak P, Raufman JP, Xiao Y and Frucht H: Functional interaction of lithocholic acid conjugates with M3 muscarinic receptors on a human colon cancer cell line. Biochim Biophys Acta. 1588:48–55. 2002. View Article : Google Scholar : PubMed/NCBI
9	Raufman JP, Chen Y, Cheng K, Compadre C, Compadre L and Zimniak P: Selective interaction of bile acids with muscarinic receptors: A case of molecular mimicry. Eur J Pharmacol. 457:77–84. 2002. View Article : Google Scholar : PubMed/NCBI
10	Shah N, Khurana S, Cheng K and Raufman JP: Muscarinic receptors and ligands in cancer. Am J Physiol Cell Physiol. 296:C221–C232. 2009. View Article : Google Scholar :
11	Trombino S, Bisio A, Catassi A, Cesario A, Falugi C and Russo P: Role of the non-neuronal human cholinergic system in lung cancer and mesothelioma: Possibility of new therapeutic strategies. Curr Med Chem Anticancer Agents. 4:535–542. 2004. View Article : Google Scholar : PubMed/NCBI
12	Español AJ, de la Torre E, Fiszman GL and Sales ME: Role of non-neuronal cholinergic system in breast cancer progression. Life Sci. 80:2281–2285. 2007. View Article : Google Scholar : PubMed/NCBI
13	Belo A, Cheng K, Chahdi A, Shant J, Xie G, Khurana S and Raufman JP: Muscarinic receptor agonists stimulate human colon cancer cell migration and invasion. Am J Physiol Gastrointest Liver Physiol. 300:G749–G760. 2011. View Article : Google Scholar : PubMed/NCBI
14	Parnell EA, Calleja-Macias IE, Kalantari M, Grando SA and Bernard HU: Muscarinic cholinergic signaling in cervical cancer cells affects cell motility via ERK1/2 signaling. Life Sci. 91:1093–1098. 2012. View Article : Google Scholar : PubMed/NCBI
15	Rayford W, Noble MJ, Austenfeld MA, Weigel J, Mebust WK and Shah GV: Muscarinic cholinergic receptors promote growth of human prostate cancer cells. Prostate. 30:160–166. 1997. View Article : Google Scholar : PubMed/NCBI
16	Paleari L, Grozio A, Cesario A and Russo P: The cholinergic system and cancer. Semin Cancer Biol. 18:211–217. 2008. View Article : Google Scholar : PubMed/NCBI
17	Schuller HM: Is cancer triggered by altered signalling of nicotinic acetylcholine receptors? Nat Rev Cancer. 9:195–205. 2009. View Article : Google Scholar : PubMed/NCBI
18	Feng YJ, Zhang BY, Yao RY and Lu Y: Muscarinic acetylcholine receptor M3 in proliferation and perineural invasion of cholangiocarcinoma cells. Hepatobiliary Pancreat Dis Int. 11:418–423. 2012. View Article : Google Scholar : PubMed/NCBI
19	Fava G, Marzioni M, Francis H, Glaser S, Demorrrow S, Ueno Y, Benedetti A and Alpini G: Novel interaction of bile acid and neural signaling in the regulation of cholangiocyte function. Hepatol Res. 37(Suppl 3): S420–S429. 2007. View Article : Google Scholar : PubMed/NCBI
20	Sirisinha S, Tengchaisri T, Boonpucknavig S, Prempracha N, Ratanarapee S and Pausawasdi A: Establishment and characterization of a cholangiocarcinoma cell line from a Thai patient with intrahepatic bile duct cancer. Asian Pac J Allergy Immunol. 9:153–157. 1991.PubMed/NCBI
21	Rattanasinganchan P, Leelawat K, Treepongkaruna SA, Tocharoentanaphol C, Subwongcharoen S, Suthiphongchai T and Tohtong R: Establishment and characterization of a cholangiocarcinoma cell line (RMCCA-1) from a Thai patient. World J Gastroenterol. 12:6500–6506. 2006.PubMed/NCBI
22	LeSage G, Alvaro D, Benedetti A, Glaser S, Marucci L, Baiocchi L, Eisel W, Caligiuri A, Phinizy JL, Rodgers R, et al: Cholinergic system modulates growth, apoptosis, and secretion of cholangiocytes from bile duct-ligated rats. Gastroenterology. 117:191–199. 1999. View Article : Google Scholar : PubMed/NCBI
23	Dai J, Wang H, Dong Y, Zhang Y and Wang J: Bile acids affect the growth of human cholangiocarcinoma via NF-κB pathway. Cancer Invest. 31:111–120. 2013. View Article : Google Scholar : PubMed/NCBI
24	Werneburg NW, Yoon JH, Higuchi H and Gores GJ: Bile acids activate EGF receptor via a TGF-alpha-dependent mechanism in human cholangiocyte cell lines. Am J Physiol Gastrointest Liver Physiol. 285:G31–G36. 2003. View Article : Google Scholar : PubMed/NCBI
25	Degirolamo C, Modica S, Palasciano G and Moschetta A: Bile acids and colon cancer: Solving the puzzle with nuclear receptors. Trends Mol Med. 17:564–572. 2011. View Article : Google Scholar : PubMed/NCBI
26	Renga B, Mencarelli A, Cipriani S, D’Amore C, Carino A, Bruno A, Francisci D, Zampella A, Distrutti E and Fiorucci S: The bile acid sensor FXR is required for immune-regulatory activities of TLR-9 in intestinal inflammation. PLoS One. 8:e544722013. View Article : Google Scholar : PubMed/NCBI
27	Zhang Y, Hong JY, Rockwell CE, Copple BL, Jaeschke H and Klaassen CD: Effect of bile duct ligation on bile acid composition in mouse serum and liver. Liver Int. 32:58–69. 2012. View Article : Google Scholar :
28	Tribe RM, Dann AT, Kenyon AP, Seed P, Shennan AH and Mallet A: Longitudinal profiles of 15 serum bile acids in patients with intrahepatic cholestasis of pregnancy. Am J Gastroenterol. 105:585–595. 2010. View Article : Google Scholar
29	Sharif AW, Williams HR, Lampejo T, Khan SA, Bansi DS, Westaby D, Thillainayagam AV, Thomas HC, Cox IJ and Taylor-Robinson SD: Metabolic profiling of bile in cholangiocarcinoma using in vitro magnetic resonance spectroscopy. HPB (Oxford). 12:396–402. 2010. View Article : Google Scholar
30	Bobeldijk I, Hekman M, de Vries-van der Weij J, Coulier L, Ramaker R, Kleemann R, Kooistra T, Rubingh C, Freidig A and Verheij E: Quantitative profiling of bile acids in biofluids and tissues based on accurate mass high resolution LC-FT-MS: Compound class targeting in a metabolomics workflow. J Chromatogr B Analyt Technol Biomed Life Sci. 871:306–313. 2008. View Article : Google Scholar : PubMed/NCBI
31	Greco AV and Mingrone G: Serum bile acid concentrations in mild liver cirrhosis. Clin Chim Acta. 221:183–189. 1993. View Article : Google Scholar : PubMed/NCBI
32	Xu ZP, Yang K, Xu GN, Zhu L, Hou LN, Zhang WH, Chen HZ and Cui YY: Role of M3 mAChR in in vivo and in vitro models of LPS-induced inflammatory response. Int Immunopharmacol. 14:320–327. 2012. View Article : Google Scholar : PubMed/NCBI
33	Sales ME: Muscarinic receptors as targets for anti-inflammatory therapy. Curr Opin Investig Drugs. 11:1239–1245. 2010.PubMed/NCBI
34	Avissar NE, Toia L, Hu Y, Watson TJ, Jones C, Raymond DP, Matousek A and Peters JH: Bile acid alone, or in combination with acid, induces CDX2 expression through activation of the epidermal growth factor receptor (EGFR). J Gastrointest Surg. 13:212–222. 2009. View Article : Google Scholar