Blocking of the EGFR‑STAT3 signaling pathway through afatinib treatment inhibited the intrahepatic cholangiocarcinoma

Zhang,Changhe; Xu,Hong; Zhou,Zhenping; Tian,Ye; Cao,Xiaofei; Cheng,Guochang; Liu,Qinghong

doi:10.3892/etm.2018.6038

Blocking of the EGFR‑STAT3 signaling pathway through afatinib treatment inhibited the intrahepatic cholangiocarcinoma

Authors:
- Changhe Zhang
- Hong Xu
- Zhenping Zhou
- Ye Tian
- Xiaofei Cao
- Guochang Cheng
- Qinghong Liu
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Affiliations: Department of Hepatobiliary Surgery, Taizhou People's Hospital, Taizhou, Jiangsu 225300, P.R. China, Department of Digestive Medicine, Taizhou People's Hospital, Taizhou, Jiangsu 225300, P.R. China
Published online on: April 5, 2018 https://doi.org/10.3892/etm.2018.6038
Pages: 4995-5000

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Abstract

Epidermal growth factor receptor (EGFR) and downstream signal transducer and activator of transcription 3 (STAT3) signaling have been extensively implicated in various human neoplasms. Recently, a novel EGFR inhibitor, known as afatinib, has exhibited broad antitumor activities in a variety of tumors. Therefore, the present study attempted to investigate the impact of this agent on intrahepatic cholangiocarcinoma (ICC). Initially, immunohistochemical assays were performed on 15 human ICC specimens and their adjacent tissues in order to assess the protein levels of phosphorylated EGFR (pEGFR) and pSTAT3. Subsequently, the human ICC cell lines JCK and OZ were exposed to different doses of afatinib, and then cell viability and apoptosis were determined by MTT assay and flow cytometry, respectively. Furthermore, immunoblotting was applied to detect any variations in the phosphorylated protein levels of EGFR and STAT3 in afatinib‑treated ICC cells. The results of the current study demonstrated that ICC specimens had evidently increased pEGFR and pSTAT3 protein levels as compared with the adjacent noncancerous tissues. Further in vitro experiments indicated that afatinib evidently blocked ICC cell growth and induced cell apoptosis. At the protein level, pEGFR and pSTAT3 were evidently attenuated by afatinib‑administration. In conclusion, the present study clearly determined that afatinib exerts an antitumor effect on ICC cells by silencing the EGFR‑STAT3 signaling pathway. This novel agent deserves further investigation as a potential therapeutic strategy for ICC.

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1	Shishir KM, Gamblin TC, Kamel I, Corona-Villalobos CP, Thomas M and Pawlik TM: Multidisciplinary approaches to intrahepatic cholangiocarcinoma. Cancer. 119:3929–3942. 2013. View Article : Google Scholar : PubMed/NCBI
2	Sriputtha S, Khuntikeo N, Promthet S and Kamsa-Ard S: Survival rate of intrahepatic cholangiocarcinoma patients after surgical treatment in Thailand. Asian Pacific J Cancer Prev. 14:1107–1110. 2013. View Article : Google Scholar
3	Robert R Jr: ErbB/HER protein-tyrosine kinases: Structures and small molecule inhibitors. Pharmacol Res. 87:42–59. 2014. View Article : Google Scholar : PubMed/NCBI
4	Bi WW, Zhang WH, Yin GH, Luo H, Wang SQ, Wang H, Li C, Yan WQ and Nie DZ: Analysis of indoleamine 2–3 dioxygenase (IDO) and EGFR co-expression in breast cancer tissue by immunohistochemistry. Asian Pac J Cancer Prev. 15:5535–5538. 2014. View Article : Google Scholar : PubMed/NCBI
5	Zhao X, Sun X and Li XL: Expression and clinical significance of STAT3, p-STAT3, and VEGF-C in small cell lung cancer. Asian Pac J Cancer Prev. 13:2873–2877. 2012. View Article : Google Scholar : PubMed/NCBI
6	Fang B: Genetic interactions of STAT3 and anticancer drug development. Cancers (Basel). 6:494–525. 2014. View Article : Google Scholar : PubMed/NCBI
7	Ung N, Putoczki TL, Stylli SS, Ng I, Mariadason JM, Chan TA, Zhu HJ and Luwor RB: Anti-EGFR therapeutic efficacy correlates directly with inhibition of STAT3 activity. Cancer Biol Ther. 15:623–632. 2014. View Article : Google Scholar : PubMed/NCBI
8	Chong CR and Jänne PA: The quest to overcome resistance to EGFR-targeted therapies in cancer. Nat Med. 19:1389–1400. 2013. View Article : Google Scholar : PubMed/NCBI
9	Ioannou N, Seddon AM, Dalgleish A, Mackintosh D and Modjtahedi H: Treatment with a combination of the ErbB (HER) family blocker afatinib and the IGF-IR inhibitor, NVP-AEW541 induces synergistic growth inhibition of human pancreatic cancer cells. BMC Cancer. 13:412013. View Article : Google Scholar : PubMed/NCBI
10	Seiwert TY, Fayette J, Cupissol D, Del Campo JM, Clement PM, Hitt R, Degardin M, Zhang W, Blackman A, Ehrnrooth E and Cohen EE: A randomized, phase II study of afatinib versus cetuximab in metastatic or recurrent squamous cell carcinoma of the head and neck. Ann Oncol. 25:1813–1820. 2014. View Article : Google Scholar : PubMed/NCBI
11	Lai WY, Chen CY, Yang SC, Wu JY, Chang CJ, Yang PC and Peck K: Overcoming EGFR T790M-based tyrosine kinase inhibitor resistance with an allele-specific DNAzyme. Mol Ther Nucleic Acids. 3:e1502014. View Article : Google Scholar : PubMed/NCBI
12	Machiels JP, Licitra LF, Haddad RI, Tahara M and Cohen EE: Rationale and design of LUX-Head & Neck 1: A randomised, Phase III trial of afatinib versus methotrexate in patients with recurrent and/or metastatic head and neck squamous cell carcinoma who progressed after platinum-based therapy. BMC Cancer. 14:4732014. View Article : Google Scholar : PubMed/NCBI
13	Harder J, Waiz O, Otto F, Geissler M, Olschewski M, Weinhold B, Blum HE, Schmitt-Graeff A and Opitz OG: EGFR and HER2 expression in advanced biliary tract cancer. World J Gastroenterol. 15:4511–4517. 2009. View Article : Google Scholar : PubMed/NCBI
14	Shen WF, Zhong W, Xu F, Kan T, Geng L, Xie F, Sui CJ and Yang JM: Clinicopathological and prognostic analysis of 429 patients with intrahepatic cholangiocarcinoma. World J Gastroenterol. 15:5976–5982. 2009. View Article : Google Scholar : PubMed/NCBI
15	Yoshikawa D, Ojima H, Iwasaki M, Hiraoka N, Kosuge T, Kasai S, Hirohashi S and Shibata T: Clinicopathological and prognostic significance of EGFR, VEGF, and HER2 expression in cholangiocarcinoma. Br J Cancer. 98:418–425. 2008. View Article : Google Scholar : PubMed/NCBI
16	Clapéron A, Mergey M, Aoudjehane L, Ho-Bouldoires TH, Wendum D, Prignon A, Merabtene F, Firrincieli D, Desbois-Mouthon C, Scatton O, et al: Hepatic myofibroblasts promote the progression of human cholangiocarcinoma through activation of epidermal growth factor receptor. Hepatology. 58:2001–2011. 2013. View Article : Google Scholar : PubMed/NCBI
17	Nanda S: Cancer: A limited role for dual EGFR and ErbB2 inhibition in cholangiocarcinoma? Nat Rev Gastroenterol Hepatol. 7:5912010. View Article : Google Scholar : PubMed/NCBI
18	Wu J, Patmore DM, Jousma E, Eaves DW, Breving K, Patel AV, Schwartz EB, Fuchs JR, Cripe TP, Stemmer-Rachamimov AO and Ratner N: EGFR-STAT3 signaling promotes formation of malignant peripheral nerve sheath tumors. Oncogene. 33:173–180. 2014. View Article : Google Scholar : PubMed/NCBI
19	Wendt MK, Balanis N, Carlin CR and Schiemann WP: STAT3 and epithelial-mesenchymal transitions in carcinomas. JAKSTAT. 3:e289752014.PubMed/NCBI
20	Qi QR and Yang ZM: Regulation and function of signal transducer and activator of transcription 3. World J Biol Chem. 5:231–239. 2014.PubMed/NCBI
21	Mobahat M, Narendran A and Riabowol K: Survivin as a preferential target for cancer therapy. Int J Mol Sci. 15:2495–2516. 2014. View Article : Google Scholar
22	Concha-Benavente F, Srivastava RM, Ferrone S and Ferris RL: EGFR-mediated tumor immunoescape: The imbalance between phosphorylated STAT1 and phosphorylated STAT3. Oncoimmunology. 2:e272152013. View Article : Google Scholar : PubMed/NCBI
23	Zhou J, Liu Y, Wang T, Zhang H, Du M, Zhang S, Wu S, Song S, Liu B, Zhang H and Jiang Z: Serum HER2 ECD level and its clinical significance in advanced breast cancer patients with different molecular subtypes. Zhonghua Yi Xue Za Zhi. 94:1384–1387. 2014.(In Chinese). PubMed/NCBI
24	Parums DV: Current status of targeted therapy in non-small cell lung cancer. Drugs Today (Barc). 50:503–525. 2014. View Article : Google Scholar : PubMed/NCBI