Interleukin‑6 induces epithelial‑mesenchymal transition in human intrahepatic biliary epithelial cells

Jiang,Gui‑Xing; Cao,Li‑Ping; Kang,Peng‑Cheng; Zhong,Xiang‑Yu; Lin,Tian‑Yu; Cui,Yun‑Fu

doi:10.3892/mmr.2015.4706

Interleukin‑6 induces epithelial‑mesenchymal transition in human intrahepatic biliary epithelial cells

Authors:
- Gui‑Xing Jiang
- Li‑Ping Cao
- Peng‑Cheng Kang
- Xiang‑Yu Zhong
- Tian‑Yu Lin
- Yun‑Fu Cui
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Affiliations: Department of Hepatopancreatobiliary Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310000, P.R. China, Department of Hepatopancreatobiliary Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
Published online on: December 22, 2015 https://doi.org/10.3892/mmr.2015.4706
Pages: 1563-1569
Copyright: © Jiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to determine the role of interleukin‑6 (IL‑6) in the epithelial‑mesenchymal transition (EMT) of human intrahepatic biliary epithelial cell (HIBEC) lines in vitro. HIBECs were stimulated with IL‑6 at concentrations of 0, 10, 20, 50 and 100 µg/l for 24 h. A wound healing and Transwell assay were performed to determine the migratory and invasive capacity of HIBECs, respectively. Following 24 h of incubation, IL-6 at 10 and 20 µg/l significantly increased the number of migrated and invaded cells (P<0.05), while stimulation with 50 and 100 µg/l IL‑6 resulted in a further increase of the migratory and invasive capacity compared to that in all other groups (P<0.05). Furthermore, reverse-transcription quantitative polymerase chain reaction and western blot analyses were used to detect the mRNA and protein expression of EMT markers E‑cadherin and vimentin in HIBECs. Decreased mRNA levels of E‑cadherin accompanied by higher mRNA levels of vimentin were observed in the 10, 20, 50, 100 µg/l IL-6 groups compared to those in the 0 µg/l group (all P<0.05). Furthermore, the protein expression of E‑cadherin was decreased, while that of vimentin was increased in the 50 and 100 µg/l IL-6 groups compared to those in the 0, 10 and 20 µg/l IL-6 groups (all P<0.05). The present study therefore indicated that IL‑6 promoted the process of EMT in HIBECs as characterized by increased migration and invasion of HIBECs and the typical changes in mRNA and protein expression of the EMT markers E‑cadherin and vimentin.

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1	Yang J and Weinberg RA: Epithelial-mesenchymal transition: At the crossroads of development and tumor metastasis. Dev Cell. 14:818–829. 2008. View Article : Google Scholar : PubMed/NCBI
2	Cannito S, Novo E, Di Bonzo LV, Busletta C, Colombatto S and Parola M: Epithelial-mesenchymal transition: From molecular mechanisms, redox regulation to implications in human health and disease. Antioxid Redox Signal. 12:1383–1430. 2010. View Article : Google Scholar
3	Thiery JP, Acloque H, Huang RY and Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell. 139:871–890. 2009. View Article : Google Scholar : PubMed/NCBI
4	Gonzalez DM and Medici D: Signaling mechanisms of the epithelial-mesenchymal transition. Sci Signal. 7:re82014. View Article : Google Scholar : PubMed/NCBI
5	Schulze F, Schardt K, Wedemeyer I, Konze E, Wendland K, Dirsch O, Töx U, Dienes HP and Odenthal M: Epithelial-mesenchymal transition of biliary epithelial cells in advanced liver fibrosis. Verh Dtsch Ges Pathol. 91:250–256. 2007.In German.
6	Mizuguchi YSS and Isse K: Molecular pathology of liver diseases. Hepatology. 59:1130–1143. 2014. View Article : Google Scholar
7	Rodés J, Benhamou JP, Blei A, Reichen J and Rizzetto M: From Basic Science to Clinical Practice. Lacy A and Francis NK: 3rd. Wiley-Blackwell; Hoboken, NJ: pp. 52–57. 2007
8	Yokoyama T, Komori A, Nakamura M, Takii Y, Kamihira T, Shimoda S, Mori T, Fujiwara S, Koyabu M, Taniguchi K, et al: Human intrahepatic biliary epithelial cells function in innate immunity by producing IL-6 and IL-8 via the TLR4-NF-kappaB and -MAPK signaling pathways. Liver Int. 26:467–476. 2006. View Article : Google Scholar : PubMed/NCBI
9	Karrar A, Broomé U, Södergren T, Jaksch M, Bergquist A, Björnstedt M and Sumitran-Holgersson S: Biliary epithelial cell antibodies link adaptive and innate immune responses in primary sclerosing cholangitis. Gastroenterology. 132:1504–1514. 2007. View Article : Google Scholar : PubMed/NCBI
10	Kawata K, Kobayashi Y, Gershwin ME and Bowlus CL: The immunophysiology and apoptosis of biliary epithelial cells: Primary biliary cirrhosis and primary sclerosing cholangitis. Clin Rev Allergy Immunol. 43:230–241. 2012. View Article : Google Scholar : PubMed/NCBI
11	Yasoshima M, Kono N, Sugawara H, Katayanagi K, Harada K and Nakanuma Y: Increased expression of interleukin-6 and tumor necrosis factor-alpha in pathologic biliary epithelial cells: In situ and culture study. Lab Invest. 78:89–100. 1998.PubMed/NCBI
12	Kishimoto T: Interleukin-6: Discovery of a pleiotropic cytokine. Arthritis Res Ther. 8(Suppl 2): S22006. View Article : Google Scholar : PubMed/NCBI
13	Park J, Tadlock L, Gores GJ and Patel T: Inhibition of interleukin 6-mediated mitogen-activated protein kinase activation attenuates growth of a cholangiocarcinoma cell line. Hepatology. 30:1128–1133. 1999. View Article : Google Scholar : PubMed/NCBI
14	Yokomuro S, Tsuji H, Lunz JG III, Sakamoto T, Ezure T, Murase N and Demetris AJ: Growth control of human biliary epithelial cells by interleukin 6, hepatocyte growth factor, transforming growth factor beta1 and activin A: Comparison of a cholangiocarcinoma cell line with primary cultures of non-neoplastic biliary epithelial cells. Hepatology. 32:26–35. 2000. View Article : Google Scholar : PubMed/NCBI
15	Sullivan NJ, Sasser AK, Axel AE, Vesuna F, Raman V, Ramirez N, Oberyszyn TM and Hall BM: Interleukin-6 induces an epithelial-mesenchymal transition phenotype in human breast cancer cells. Oncogene. 28:2940–2947. 2009. View Article : Google Scholar : PubMed/NCBI
16	Xie G, Yao Q, Liu Y, Du S, Liu A, Guo Z, Sun A, Ruan J, Chen L, Ye C and Yuan Y: IL-6-induced epithelial-mesenchymal transition promotes the generation of breast cancer stem-like cells analogous to mammosphere cultures. Int J Oncol. 40:1171–1179. 2012.
17	Yadav A, Kumar B, Datta J, Teknos TN and Kumar P: IL-6 promotes head and neck tumor metastasis by inducing epithelial-mesenchymal transition via the JAK-STAT3-SNAIL signaling pathway. Mol Cancer Res. 9:1658–1667. 2011. View Article : Google Scholar : PubMed/NCBI
18	Colomiere M, Ward AC, Riley C, Trenerry MK, Cameron-Smith D, Findlay J, Ackland L and Ahmed N: Cross talk of signals between EGFR and IL-6R through JAK2/STAT3 mediate epithelial-mesenchymal transition in ovarian carcinomas. Br J Cancer. 100:134–144. 2009. View Article : Google Scholar :
19	Chun J and Kim YS: Platycodin D inhibits migration, invasion, and growth of MDA-MB-231 human breast cancer cells via suppression of EGFR-mediated Akt and MAPK pathways. Chem Biol Interact. 205:212–221. 2013. View Article : Google Scholar : PubMed/NCBI
20	Causier B and Davies B: Analysing protein-protein interactions with the yeast two-hybrid system. Plant Mol Biol. 50:855–870. 2002. View Article : Google Scholar
21	Lamireau T, Zoltowska M, Levy E, Yousef I, Rosenbaum J, Tuchweber B and Desmoulière A: Effects of bile acids on biliary epithelial cells: Proliferation, cytotoxicity, and cytokine secretion. Life Sci. 72:1401–1411. 2003. View Article : Google Scholar : PubMed/NCBI
22	Ezure T, Sakamoto T, Tsuji H, Lunz JG III, Murase N, Fung JJ and Demetris AJ: The development and compensation of biliary cirrhosis in interleukin-6-deficient mice. Am J Pathol. 156:1627–1639. 2000. View Article : Google Scholar : PubMed/NCBI
23	Liu Z, Sakamoto T, Ezure T, Yokomuro S, Murase N, Michalopoulos G and Demetris AJ: Interleukin-6, hepatocyte growth factor, and their receptors in biliary epithelial cells during a type I ductular reaction in mice: Interactions between the periductal inflammatory and stromal cells and the biliary epithelium. Hepatology. 28:1260–1268. 1998. View Article : Google Scholar : PubMed/NCBI
24	Yokomuro S, Lunz JG III, Sakamoto T, Ezure T, Murase N and Demetris AJ: The effect of interleukin-6 (IL-6)/gp130 signalling on biliary epithelial cell growth, in vitro. Cytokine. 12:727–730. 2000. View Article : Google Scholar : PubMed/NCBI
25	Nozaki I, Lunz JG III, Specht S, Park JI, Giraud AS, Murase N and Demetris AJ: Regulation and function of trefoil factor family 3 expression in the biliary tree. Am J Pathol. 165:1907–1920. 2004. View Article : Google Scholar : PubMed/NCBI
26	Chen LP, Cai M, Zhang QH, Li ZL, Qian YY, Bai HW, Wei X, Shi BY and Dong JH: Activation of interleukin-6/STAT3 in rat cholangiocyte proliferation induced by lipopolysaccharide. Dig Dis Sci. 54:547–554. 2009. View Article : Google Scholar
27	Chen LP, Qian YY, Li ZL, Bai HW, Cai M and Shi BY: Role of IL-6/STAT3 in rat cholangiocyte proliferation induced by lipopolysaccharide. Zhonghua Gan Zang Bing Za Zhi. 17:374–377. 2009.In Chinese. PubMed/NCBI
28	Jiang GX, Zhong XY, Cui YF, Liu W, Tai S, Wang ZD, Shi YG, Zhao SY and Li CL: IL-6/STAT3/TFF3 signaling regulates human biliary epithelial cell migration and wound healing in vitro. Mol Biol Rep. 37:3813–3818. 2010. View Article : Google Scholar : PubMed/NCBI
29	Zeisberg M and Neilson EG: Biomarkers for epithelial-mesenchymal transitions. J Clin Invest. 119:1429–1437. 2009. View Article : Google Scholar : PubMed/NCBI
30	Kalluri R and Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 119:1420–1428. 2009. View Article : Google Scholar : PubMed/NCBI
31	Kalluri R and Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest. 112:1776–1784. 2003. View Article : Google Scholar : PubMed/NCBI
32	Huber MA, Kraut N and Beug H: Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol. 17:548–558. 2005. View Article : Google Scholar : PubMed/NCBI
33	Kokkinos MI, Wafai R, Wong MK, Newgreen DF, Thompson EW and Waltham M: Vimentin and epithelial-mesenchymal transition in human breast cancer-observations in vitro and in vivo. Cells Tissues Organs. 185:191–203. 2007. View Article : Google Scholar
34	Lee JM, Dedhar S, Kalluri R and Thompson EW: The epithelial-mesenchymal transition: New insights in signaling, development, and disease. J Cell Biol. 172:973–981. 2006. View Article : Google Scholar : PubMed/NCBI
35	Mendez MG, Kojima S and Goldman RD: Vimentin induces changes in cell shape, motility, and adhesion during the epithelial to mesenchymal transition. FASEB J. 24:1838–1851. 2010. View Article : Google Scholar : PubMed/NCBI