Cx32 reverses epithelial-mesenchymal transition in doxorubicin-resistant hepatocellular carcinoma

Yu,Meiling; Han,Guangshu; Qi,Benquan; Wu,Xiaoxiang

doi:10.3892/or.2017.5462

Cx32 reverses epithelial-mesenchymal transition in doxorubicin-resistant hepatocellular carcinoma

Authors:
- Meiling Yu
- Guangshu Han
- Benquan Qi
- Xiaoxiang Wu
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Affiliations: Department of Pharmacy, The First Affiliated Hospital of Bengbu Medical College, Anhui, Bengbu 233004, P.R. China, Faculty of Pharmacy, Bengbu Medical College, Anhui, Bengbu 233030, P.R. China, Department of Emergency Internal Medicine, The First Affiliated Hospital of Bengbu Medical College, Anhui, Bengbu 233004, P.R. China, Department of Pharmacy, The Second Affiliated Hospital of Bengbu Medical College, Anhui, Bengbu 233004, P.R. China
Published online on: February 17, 2017 https://doi.org/10.3892/or.2017.5462
Pages: 2121-2128

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Abstract

Recently, epithelial-mesenchymal transition (EMT) has been reported to be an important mechanism of drug resistance in numerous types of cancer cells, including hepatocellular carcinoma (HCC). However, the underlying mechanisms remain to be fully elucidated. Connexin (Cx)32 plays a crucial role in hepatocarcinogenesis. The present study investigated the role of Cx32 in the regulation of chemotherapy-induced EMT in HCC. We found that the expression levels of Cx32 and E-cadherin were clearly decreased in HCC tissues compared with the corresponding paracancerous tissues, while the expression level of vimentin was significantly enhanced in HCC tissues. The expression of Cx32 had a strong correlation with the expression of E-cadherin and vimentin. In an in vitro study, a doxorubicin (DOX)-resistant liver cell line HepG2/DOX was established from parental HepG2 cells. The results showed that HepG2/DOX cells acquired EMT characteristics, with a decreased expression level of E-cadherin and an enhanced expression level of vimentin, and possessed high migratory abilities and invasiveness. Meanwhile, Cx32 was significantly decreased in the HepG2/DOX cells. Knockdown of Cx32 by shRNA in HepG2 cells induced EMT, while overexpression of Cx32 converted EMT to mesenchymal-epithelial transition (MET) in the HepG2/DOX cells. These results suggest that Cx32 is an important regulator of DOX-induced EMT in HCC. Cx32 could be considered as a novel target to reverse DOX resistance in HCC.

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1	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
2	Zhu AX: Systemic therapy of advanced hepatocellular carcinoma: How hopeful should we be? Oncologist. 11:790–800. 2006. View Article : Google Scholar : PubMed/NCBI
3	Asghar U and Meyer T: Are there opportunities for chemotherapy in the treatment of hepatocellular cancer? J Hepatol. 56:686–695. 2012. View Article : Google Scholar : PubMed/NCBI
4	Xu F, Wang F, Yang T, Sheng Y, Zhong T and Chen Y: Differential drug resistance acquisition to doxorubicin and paclitaxel in breast cancer cells. Cancer Cell Int. 14:5382014. View Article : Google Scholar : PubMed/NCBI
5	Harada K, Ferdous T and Ueyama Y: Establishment of 5-fluorouracil-resistant oral squamous cell carcinoma cell lines with epithelial to mesenchymal transition changes. Int J Oncol. 44:1302–1308. 2014.PubMed/NCBI
6	Haider M, Zhang X, Coleman I, Ericson N, True LD, Lam HM, Brown LG, Ketchanji M, Nghiem B, Lakely B, et al: Epithelial mesenchymal-like transition occurs in a subset of cells in castration resistant prostate cancer bone metastases. Clin Exp Metastasis. 33:239–248. 2016. View Article : Google Scholar : PubMed/NCBI
7	Xu H, Pirisi L and Creek KE: Six1 overexpression at early stages of HPV16-mediated transformation of human keratinocytes promotes differentiation resistance and EMT. Virology. 474:144–153. 2015. View Article : Google Scholar : PubMed/NCBI
8	Dinicola S, Pasqualato A, Proietti S, Masiello MG, Palombo A, Coluccia P, Canipari R, Catizone A, Ricci G, Harrath AH, et al: Paradoxical E-cadherin increase in 5FU-resistant colon cancer is unaffected during mesenchymal-epithelial reversion induced by γ-secretase inhibition. Life Sci. 145:174–183. 2016. View Article : Google Scholar : PubMed/NCBI
9	Zhou JZ and Jiang JX: Gap junction and hemichannel-independent actions of connexins on cell and tissue functions - an update. FEBS Lett. 588:1186–1192. 2014. View Article : Google Scholar : PubMed/NCBI
10	Aasen T: Connexins: Junctional and non-junctional modulators of proliferation. Cell Tissue Res. 360:685–699. 2015. View Article : Google Scholar : PubMed/NCBI
11	Jiang JX and Gu S: Gap junction- and hemichannel-independent actions of connexins. Biochim Biophys Acta. 1711:208–214. 2005. View Article : Google Scholar : PubMed/NCBI
12	Diezmos EF, Bertrand PP and Liu L: Purinergic signaling in gut inflammation: The role of connexins and pannexins. Front Neurosci. 10:3112016. View Article : Google Scholar : PubMed/NCBI
13	Cronier L, Crespin S, Strale PO, Defamie N and Mesnil M: Gap junctions and cancer: New functions for an old story. Antioxid Redox Signal. 11:323–338. 2009. View Article : Google Scholar : PubMed/NCBI
14	Kandouz M and Batist G: Gap junctions and connexins as therapeutic targets in cancer. Expert Opin Ther Targets. 14:681–692. 2010. View Article : Google Scholar : PubMed/NCBI
15	Trosko JE and Ruch RJ: Gap junctions as targets for cancer chemoprevention and chemotherapy. Curr Drug Targets. 0.3:465–482. 2002. View Article : Google Scholar
16	Yu M, Zhang C, Li L, Dong S, Zhang N and Tong X: Cx43 reverses the resistance of A549 lung adenocarcinoma cells to cisplatin by inhibiting EMT. Oncol Rep. 31:2751–2758. 2014.PubMed/NCBI
17	Song J and Li Y: miR-25-3p reverses epithelial-mesenchymal transition via targeting Sema4C in cisplatin-resistance cervical cancer cells. Cancer Sci. Oct 15–2016.(Epub ahead of print). doi: 10.1111/cas.13104.
18	Gao HX, Yan L, Li C, Zhao LM and Liu W: miR-200c regulates crizotinib-resistant ALK-positive lung cancer cells by reversing epithelial-mesenchymal transition via targeting ZEB1. Mol Med Rep. 14:4135–4143. 2016.PubMed/NCBI
19	Wu Q, Wang R, Yang Q, Hou X, Chen S, Hou Y, Chen C, Yang Y, Miele L, Sarkar FH, et al: Chemoresistance to gemcitabine in hepatoma cells induces epithelial-mesenchymal transition and involves activation of PDGF-D pathway. Oncotarget. 4:1999–2009. 2013. View Article : Google Scholar : PubMed/NCBI
20	Hiscox S, Jiang WG, Obermeier K, Taylor K, Morgan L, Burmi R, Barrow D and Nicholson RI: Tamoxifen resistance in MCF7 cells promotes EMT-like behaviour and involves modulation of β-catenin phosphorylation. Int J Cancer. 118:290–301. 2006. View Article : Google Scholar : PubMed/NCBI
21	Du B and Shim JS: Targeting epithelial-mesenchymal transition (EMT) to overcome drug resistance in cancer. Molecules. 21:212016. View Article : Google Scholar
22	Kay CW, Ursu D, Sher E and King AE: The role of Cx36 and Cx43 in 4-aminopyridine-induced rhythmic activity in the spinal nociceptive dorsal horn: An electrophysiological study in vitro. Physiol Rep. 4:42016. View Article : Google Scholar
23	Unger VM, Kumar NM, Gilula NB and Yeager M: Three-dimensional structure of a recombinant gap junction membrane channel. Science. 283:1176–1180. 1999. View Article : Google Scholar : PubMed/NCBI
24	Maes M, Decrock E, Cogliati B, Oliveira AG, Marques PE, Dagli ML, Menezes GB, Mennecier G, Leybaert L, Vanhaecke T, et al: Connexin and pannexin (hemi)channels in the liver. Front Physiol. 4:4052014. View Article : Google Scholar : PubMed/NCBI
25	De Maio A, Gingalewski C, Theodorakis NG and Clemens MG: Interruption of hepatic gap junctional communication in the rat during inflammation induced by bacterial lipopolysaccharide. Shock. 14:53–59. 2000. View Article : Google Scholar : PubMed/NCBI
26	Sagawa H, Naiki-Ito A, Kato H, Naiki T, Yamashita Y, Suzuki S, Sato S, Shiomi K, Kato A, Kuno T, et al: Connexin 32 and luteolin play protective roles in non-alcoholic steatohepatitis development and its related hepatocarcinogenesis in rats. Carcinogenesis. 36:1539–1549. 2015.PubMed/NCBI
27	Nakashima Y, Ono T, Yamanoi A, El-Assal ON, Kohno H and Nagasue N: Expression of gap junction protein connexin32 in chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. J Gastroenterol. 39:763–768. 2004. View Article : Google Scholar : PubMed/NCBI
28	Zhao B, Zhao W, Wang Y, Xu Y, Xu J, Tang K, Zhang S, Yin Z, Wu Q and Wang X: Connexin32 regulates hepatoma cell metastasis and proliferation via the p53 and Akt pathways. Oncotarget. 6:10116–10133. 2015. View Article : Google Scholar : PubMed/NCBI
29	Yang J, Ichikawa A and Tsuchiya T: A novel function of connexin 32: Marked enhancement of liver function in a hepatoma cell line. Biochem Biophys Res Commun. 307:80–85. 2003. View Article : Google Scholar : PubMed/NCBI
30	Muramatsu A, Iwai M, Morikawa T, Tanaka S, Mori T, Harada Y and Okanoue T: Influence of transfection with connexin 26 gene on malignant potential of human hepatoma cells. Carcinogenesis. 23:351–358. 2002. View Article : Google Scholar : PubMed/NCBI
31	Decrock E, Vinken M, De Vuyst E, Krysko DV, Herde DK, Vanhaecke T, Vandenabeele P, Rogiers V and Leybaert L: Connexin-related signaling in cell death: To live or let die? Cell Death Differ. 16:524–536. 2009. View Article : Google Scholar : PubMed/NCBI
32	Chandrasekhar A and Bera AK: Hemichannels: Permeants and their effect on development, physiology and death. Cell Biochem Funct. 30:89–100. 2012. View Article : Google Scholar : PubMed/NCBI
33	Kar R, Batra N, Riquelme MA and Jiang JX: Biological role of connexin intercellular channels and hemichannels. Arch Biochem Biophys. 524:2–15. 2012. View Article : Google Scholar : PubMed/NCBI
34	Unahabhokha T, Chanvorachote P, Sritularak B, Kitsongsermthon J and Pongrakhananon V: Gigantol Inhibits Epithelial to Mesenchymal Process in Human Lung Cancer Cells. Evid Based Complement Alternat Med. 2016:45616742016. View Article : Google Scholar : PubMed/NCBI
35	Park SJ, Choi YS, Lee S, Lee YJ, Hong S, Han S and Kim BC: BIX02189 inhibits TGF-β1-induced lung cancer cell metastasis by directly targeting TGF-β type I receptor. Cancer Lett. 381:314–322. 2016. View Article : Google Scholar : PubMed/NCBI
36	Zhou Q, Abraham AD, Li L, Babalmorad A, Bagby S, Arcaroli JJ, Hansen RJ, Valeriote FA, Gustafson DL, Schaack J, et al: Topoisomerase IIα mediates TCF-dependent epithelial-mesenchymal transition in colon cancer. Oncogene. 35:4990–4999. 2016. View Article : Google Scholar : PubMed/NCBI
37	Wang HJ, Zhu J and Zheng GY: Role of glutathione and other antioxidants in the inhibition of apoptosis and mesenchymal transition in rabbit lens epithelial cells. Genet Mol Res. 13:7149–7156. 2014. View Article : Google Scholar : PubMed/NCBI
38	Zhang J, Qiu M, Ma Y, Bu Y, Yang L and Tang X: Advanced oxidation protein products induce epithelial-to-mesenchymal transition in cultured human proximal tubular epithelial cells via oxidative stress. Nan Fang Yi Ke Da Xue Xue Bao. 34:659–663. 2014.(In Chinese). PubMed/NCBI
39	Ryoo IG, Shin DH, Kang KS and Kwak MK: Involvement of Nrf2-GSH signaling in TGFβ1-stimulated epithelial-to-mesenchymal transition changes in rat renal tubular cells. Arch Pharm Res. 38:272–281. 2015. View Article : Google Scholar : PubMed/NCBI