Tramadol attenuates the sensitivity of glioblastoma to temozolomide through the suppression of Cx43‑mediated gap junction intercellular communication

Wang,Lingzhi; Peng,Yuexia; Peng,Jianxin; Shao,Min; Ma,Li; Zhu,Zhuoli; Zhong,Guocheng; Xia,Zhengyuan; Huang,Huansen

doi:10.3892/ijo.2017.4188

Tramadol attenuates the sensitivity of glioblastoma to temozolomide through the suppression of Cx43‑mediated gap junction intercellular communication

Authors:
- Lingzhi Wang
- Yuexia Peng
- Jianxin Peng
- Min Shao
- Li Ma
- Zhuoli Zhu
- Guocheng Zhong
- Zhengyuan Xia
- Huansen Huang
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Affiliations: Department of Anaesthesia, The Second Affiliated Hospital, Guangzhou Medical University, Guangzhou, Guangdong 510260, P.R. China, Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong 510080, P.R. China, Department of Hepatobiliary Surgery, Guangdong Province Traditional Chinese Medicine Hospital, Guangzhou, Guangdong 510120, P.R. China, Department of Cardiovascular Internal Medicine, Tianyou Hospital, Wuhan University of Science and Technology, Wuhan, Hubei 430064, P.R. China
Published online on: November 3, 2017 https://doi.org/10.3892/ijo.2017.4188
Pages: 295-304

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Abstract

Analgesics and antineoplastic drugs are often used concurrently for cancer patients. Our previous study reported that gap junctions composed of connexin32 (Cx32) was implicated in the effect of analgesics on cisplatin cytotoxicity. However, the effect of analgesic on the most widely expressed connexin (Cx), connexin43 (Cx43), and whether such effect mediates the influence on chemotherapeutic efficiency remain unknown. By manipulation of Cx43 expression or gap junction function, we found that there were gap junction-dependent and independent effect of Cx43 on temozolomide (TMZ) sensitivity in U87 glioblastoma cells. Studies on survival and apoptosis showed widely used analgesic tramadol significantly reduced TMZ-induced cytotoxicity in control and negative control cells but not shCx43-transfected cells. Proliferation assay demonstrated tramadol suppressed TMZ-induced cytotoxicity only on high density (with gap junction formation) but not on low density (without gap junction formation). Tramadol inhibited dye-coupling through gap junctions between U87 cells. Tramadol treatment for 72 h did not alter Cx43 expression, but decreased Cx43 phosphorylation accompanied with reduced p-ERK and p-JNK. Our results indicated that long-term treatment with tramadol reduced TMZ cytotoxicity in U87 cells by suppressing Cx43-composed gap junctions, suggesting identification and usage of antinociceptive drugs which do not downregulate connexin activity should have beneficial therapeutic consequences.

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1	Leppert W, Zajaczkowska R, Wordliczek J, Dobrogowski J, Woron J and Krzakowski M: Pathophysiology and clinical characteristics of pain in most common locations in cancer patients. J Physiol Pharmacol. 67:787–799. 2016.
2	Grek CL, Rhett JM, Bruce JS, Ghatnekar GS and Yeh ES: Connexin 43, breast cancer tumor suppressor: Missed connections? Cancer Lett. 374:117–126. 2016. View Article : Google Scholar : PubMed/NCBI
3	Willebrords J, Crespo Yanguas S, Maes M, Decrock E, Wang N, Leybaert L, da Silva TC, Veloso Alves Pereira I, Jaeschke H, Cogliati B, et al: Structure, regulation and function of gap junctions in liver. Cell Commun Adhes. 22:29–37. 2015. View Article : Google Scholar
4	Hong X, Wang Q, Yang Y, Zheng S, Tong X, Zhang S, Tao L and Harris AL: Gap junctions propagate opposite effects in normal and tumor testicular cells in response to cisplatin. Cancer Lett. 317:165–171. 2012. View Article : Google Scholar
5	Jensen R and Glazer PM: Cell-interdependent cisplatin killing by Ku/DNA-dependent protein kinase signaling transduced through gap junctions. Proc Natl Acad Sci USA. 101:6134–6139. 2004. View Article : Google Scholar : PubMed/NCBI
6	Xiao J, Zhang G, Li B, Wu Y, Liu X, Tan Y and Du B: Dioscin augments HSV-tk-mediated suicide gene therapy for melanoma by promoting connexin-based intercellular communication. Oncotarget. 8:798–807. 2017.
7	Kim YJ, Kim J, Tian C, Lim HJ, Kim YS, Chung JH and Choung YH: Prevention of cisplatin-induced ototoxicity by the inhibition of gap junctional intercellular communication in auditory cells. Cell Mol Life Sci. 71:3859–3871. 2014. View Article : Google Scholar : PubMed/NCBI
8	Wang L, Fu Y, Peng J, Wu D, Yu M, Xu C, Wang Q and Tao L: Simvastatin-induced up-regulation of gap junctions composed of connexin 43 sensitize Leydig tumor cells to etoposide: An involvement of PKC pathway. Toxicology. 312:149–157. 2013. View Article : Google Scholar : PubMed/NCBI
9	Sin WC, Aftab Q, Bechberger JF, Leung JH, Chen H and Naus CC: Astrocytes promote glioma invasion via the gap junction protein connexin43. Oncogene. 35:1504–1516. 2016. View Article : Google Scholar
10	Plante I, Stewart MK, Barr K, Allan AL and Laird DW: Cx43 suppresses mammary tumor metastasis to the lung in a Cx43 mutant mouse model of human disease. Oncogene. 30:1681–1692. 2011. View Article : Google Scholar
11	He B, Tong X, Wang L, Wang Q, Ye H, Liu B, Hong X, Tao L and Harris AL: Tramadol and flurbiprofen depress the cytotoxicity of cisplatin via their effects on gap junctions. Clin Cancer Res. 15:5803–5810. 2009. View Article : Google Scholar : PubMed/NCBI
12	Suzhi Z, Liang T, Yuexia P, Lucy L, Xiaoting H, Yuan Z and Qin W: Gap junctions enhance the antiproliferative effect of MicroRNA-124-3p in glioblastoma cells. J Cell Physiol. 230:2476–2488. 2015. View Article : Google Scholar : PubMed/NCBI
13	Cottin S, Gould PV, Cantin L and Caruso M: Gap junctions in human glioblastomas: Implications for suicide gene therapy. Cancer Gene Ther. 18:674–681. 2011. View Article : Google Scholar : PubMed/NCBI
14	Goldberg GS, Bechberger JF and Naus CC: A pre-loading method of evaluating gap junctional communication by fluorescent dye transfer. Biotechniques. 18:490–497. 1995.PubMed/NCBI
15	Koreen IV, Elsayed WA, Liu YJ and Harris AL: Tetracycline-regulated expression enables purification and functional analysis of recombinant connexin channels from mammalian cells. Biochem J. 383:111–119. 2004. View Article : Google Scholar : PubMed/NCBI
16	Su J, Cai M, Li W, Hou B, He H, Ling C, Huang T, Liu H and Guo Y: Molecularly targeted drugs plus radiotherapy and temozolomide treatment for newly diagnosed glioblastoma: A meta-analysis and systematic review. Oncol Res. 24:117–128. 2016. View Article : Google Scholar : PubMed/NCBI
17	Deng L, Ng L, Ozawa T and Stella N: Quantitative analyses of synergistic responses between cannabidiol and DNA-damaging agents on the proliferation and viability of glioblastoma and neural progenitor cells in culture. J Pharmacol Exp Ther. 360:215–224. 2017. View Article : Google Scholar
18	Autsavapromporn N, Suzuki M, Funayama T, Usami N, Plante I, Yokota Y, Mutou Y, Ikeda H, Kobayashi K, Kobayashi Y, et al: Gap junction communication and the propagation of bystander effects induced by microbeam irradiation in human fibroblast cultures: The impact of radiation quality. Radiat Res. 180:367–375. 2013. View Article : Google Scholar : PubMed/NCBI
19	Lawrence TS, Rehemtulla A, Ng EY, Wilson M, Trosko JE and Stetson PL: Preferential cytotoxicity of cells transduced with cytosine deaminase compared to bystander cells after treatment with 5-flucytosine. Cancer Res. 58:2588–2593. 1998.PubMed/NCBI
20	Zhang Y, Tao L, Fan L, Peng Y, Yang K, Zhao Y, Song Q and Wang Q: Different gap junction-propagated effects on cisplatin transfer result in opposite responses to cisplatin in normal cells versus tumor cells. Sci Rep. 5:125632015. View Article : Google Scholar : PubMed/NCBI
21	Zhang Y, Wang X, Wang Q, Ge H and Tao L: Propofol depresses cisplatin cytotoxicity via the inhibition of gap junctions. Mol Med Rep. 13:4715–4720. 2016. View Article : Google Scholar : PubMed/NCBI
22	Murphy SF, Varghese RT, Lamouille S, Guo S, Pridham KJ, Kanabur P, Osimani AM, Sharma S, Jourdan J, Rodgers CM, et al: Connexin 43 inhibition sensitizes chemoresistant glioblastoma cells to temozolomide. Cancer Res. 76:139–149. 2016. View Article : Google Scholar :
23	Gielen PR, Aftab Q, Ma N, Chen VC, Hong X, Lozinsky S, Naus CC and Sin WC: Connexin43 confers Temozolomide resistance in human glioma cells by modulating the mitochondrial apoptosis pathway. Neuropharmacology. 75:539–548. 2013. View Article : Google Scholar : PubMed/NCBI
24	Naiki-Ito A, Asamoto M, Naiki T, Ogawa K, Takahashi S, Sato S and Shirai T: Gap junction dysfunction reduces acetaminophen hepatotoxicity with impact on apoptotic signaling and connexin 43 protein induction in rat. Toxicol Pathol. 38:280–286. 2010. View Article : Google Scholar : PubMed/NCBI
25	Kameritsch P, Pogoda K and Pohl U: Channel-independent influence of connexin 43 on cell migration. Biochim Biophys Acta. 1818:1993–2001. 2012. View Article : Google Scholar
26	Grond S and Sablotzki A: Clinical pharmacology of tramadol. Clin Pharmacokinet. 43:879–923. 2004. View Article : Google Scholar : PubMed/NCBI
27	Edmondson MA, Duran SH, Boothe DM, Stewart AJ and Ravis WR: Pharmacokinetics of tramadol and its major metabolites in alpacas following intravenous and oral administration. J Vet Pharmacol Ther. 35:389–396. 2012. View Article : Google Scholar
28	Li C, Chen SQ, Chen BX, Huang WQ and Liu KX: The antinociceptive effect of intrathecal tramadol in rats: The role of alpha 2-adrenoceptors in the spinal cord. J Anesth. 26:230–235. 2012. View Article : Google Scholar
29	Xia M, Tong JH, Zhou ZQ, Duan ML, Xu JG, Zeng HJ and Wang SH: Tramadol inhibits proliferation, migration and invasion via α2-adrenoceptor signaling in breast cancer cells. Eur Rev Med Pharmacol Sci. 20:157–165. 2016.
30	Sheikholeslami B, Gholami M, Lavasani H and Rouini M: Evaluation of the route dependency of the pharmacokinetics and neuro-pharmacokinetics of tramadol and its main metabolites in rats. Eur J Pharm Sci. 92:55–63. 2016. View Article : Google Scholar : PubMed/NCBI
31	Pergolizzi JV Jr, Taylor R Jr and Raffa RB: Extended-release formulations of tramadol in the treatment of chronic pain. Expert Opin Pharmacother. 12:1757–1768. 2011. View Article : Google Scholar : PubMed/NCBI
32	Leppert W and Mikolajczak P: Analgesic effects and assays of controlled-release tramadol and o-desmethyltramadol in cancer patients with pain. Curr Pharm Biotechnol. 12:306–312. 2011. View Article : Google Scholar
33	Pan EC, Bohn LM, Belcheva MM, Thomas GE, Manepalli AN, Mamone JY, Johnson FE and Coscia CJ: Kappa-opioid receptor binding varies inversely with tumor grade in human gliomas. Cancer. 83:2561–2566. 1998. View Article : Google Scholar
34	Solan JL and Lampe PD: Connexin43 phosphorylation: Structural changes and biological effects. Biochem J. 419:261–272. 2009. View Article : Google Scholar : PubMed/NCBI
35	Zhao Y, Yu L, Xu S, Qiu F, Fan Y and Fu G: Down-regulation of connexin43 gap junction by serum deprivation in human endothelial cells was improved by (-)-epigallocatechin gallate via ERK MAP kinase pathway. Biochem Biophys Res Commun. 404:217–222. 2011. View Article : Google Scholar
36	TenBroek EM, Lampe PD, Solan JL, Reynhout JK and Johnson RG: Ser364 of connexin43 and the upregulation of gap junction assembly by cAMP. J Cell Biol. 155:1307–1318. 2001. View Article : Google Scholar
37	Mathews J and Levin M: Gap junctional signaling in pattern regulation: Physiological network connectivity instructs growth and form. Dev Neurobiol. 77:643–673. 2016. View Article : Google Scholar : PubMed/NCBI
38	Brockmeyer P, Jung K, Perske C, Schliephake H and Hemmerlein B: Membrane connexin 43 acts as an independent prognostic marker in oral squamous cell carcinoma. Int J Oncol. 45:273–281. 2014. View Article : Google Scholar : PubMed/NCBI
39	Zhang A, Hitomi M, Bar-Shain N, Dalimov Z, Ellis L, Velpula KK, Fraizer GC, Gourdie RG and Lathia JD: Connexin 43 expression is associated with increased malignancy in prostate cancer cell lines and functions to promote migration. Oncotarget. 6:11640–11651. 2015. View Article : Google Scholar : PubMed/NCBI
40	Czyz J: The stage-specific function of gap junctions during tumourigenesis. Cell Mol Biol Lett. 13:92–102. 2008. View Article : Google Scholar
41	Chen MJ, Kress B, Han X, Moll K, Peng W, Ji RR and Nedergaard M: Astrocytic CX43 hemichannels and gap junctions play a crucial role in development of chronic neuropathic pain following spinal cord injury. Glia. 60:1660–1670. 2012. View Article : Google Scholar : PubMed/NCBI