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Cisplatin Activates Volume-Sensitive Like Chloride Channels Via Purinergic Receptor Pathways in Nasopharyngeal Carcinoma Cells

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Abstract

Cisplatin-based concomitant chemoradiotherapy is considered as the standard treatment for locally advanced nasopharyngeal carcinoma patients. However, the curative efficacy of cisplatin-based chemotherapy is limited because of the occurrence of cisplatin resistance. Some researches indicate that activating the volume-sensitive Cl channel might be a new strategy for the reduction of cisplatin resistance. However, little is known about the activation pathway of the Cl channels activated by cisplatin. In this study, the cisplatin-activated chloride current was investigated using the whole cell patch-clamp technique in the poorly differentiated nasopharyngeal carcinoma cells (CNE-2Z cells), and the activation pathway of the current was also discussed. The results showed that extracellular application of cisplatin activated a Cl current, showing the properties of significant outward rectification, intracellular ATP dependency, and a selectivity sequence of I > Br > Cl > gluconate, and being inhibited by the Cl channel inhibitors tamoxifen and extracellular ATP. These characteristics are similar to those of the volume-sensitive Cl current in CNE-2Z cells, indicating that cisplatin induces the Cl current by activating the volume-sensitive like chloride channel. The cisplatin-activated current was blocked by suramin (a wide-spectrum purinergic antagonist) and RB2 (a relatively selective P2Y antagonist). In addition, the current was depressed by extracellular application of apyrase. The apoptotic volume decrease induced by cisplatin was also attenuated by RB2. P2Y receptors were expressed in CNE-2Z cells. These results suggest that cisplatin can induce a Cl current by activating volume-sensitive like Cl channels through the P2Y purinoceptor pathway.

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References

  • Almeida LO, Abrahao AC, Rosselli-Murai LK, Giudice FS, Zagni C, Leopoldino AM, Squarize CH, Castilho RM (2014) NFkappaB mediates cisplatin resistance through histone modifications in head and neck squamous cell carcinoma (HNSCC). FEBS Open Bio 4:96–104. doi:10.1016/j.fob.2013.12.003

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Boarder MR, Hourani SM (1998) The regulation of vascular function by P2 receptors: multiple sites and multiple receptors. Trends Pharmacol Sci 19:99–107

    Article  CAS  PubMed  Google Scholar 

  • Chen L, Wang L, Zhu L, Nie S, Zhang J, Zhong P, Cai B, Luo H, Jacob TJ (2002) Cell cycle-dependent expression of volume-activated chloride currents in nasopharyngeal carcinoma cells. Am J Physiol Cell Physiol 283:C1313–C1323. doi:10.1152/ajpcell.00182.2002

    Article  CAS  PubMed  Google Scholar 

  • Chen L, Zhu L, Jacob TJ, Wang L (2007) Roles of volume-activated Cl currents and regulatory volume decrease in the cell cycle and proliferation in nasopharyngeal carcinoma cells. Cell Prolif 40:253–267. doi:10.1111/j.1365-2184.2007.00432.x

    Article  CAS  PubMed  Google Scholar 

  • Darby M, Kuzmiski JB, Panenka W, Feighan D, MacVicar BA (2003) ATP released from astrocytes during swelling activates chloride channels. J Neurophysiol 89:1870–1877. doi:10.1152/jn.00510.2002

    Article  CAS  PubMed  Google Scholar 

  • Fan A, Wang L, Mao J, Yang X, Nie S, Chen L (2007) Role of chloride channels in apoptotic volume decrease of nasopharyngeal carcinoma cells induced by cDDP. Shandong Med J 47:40–42 (in Chinese)

    CAS  Google Scholar 

  • Feranchak AP, Fitz JG, Roman RM (2000) Volume-sensitive purinergic signaling in human hepatocytes. J Hepatol 33:174–182

    Article  CAS  PubMed  Google Scholar 

  • He Q, Wang L, Mao J, Sun X, Li P, Zhong P, Nie S, Jacob TJ, Chen L (2004) Activation of chloride current and decrease of cell volume by ATP in nasopharyngeal carcinoma cells. Sheng Li Xue Bao 56:691–696

    CAS  PubMed  Google Scholar 

  • Ise T, Shimizu T, Lee EL, Inoue H, Kohno K, Okada Y (2005) Roles of volume-sensitive Cl channel in cisplatin-induced apoptosis in human epidermoid cancer cells. J Membr Biol 205:139–145. doi:10.1007/s00232-005-0779-y

    Article  CAS  PubMed  Google Scholar 

  • Jin Y, Cai X, Shi Y, Xia X, Cai Y, Cao Y, Zhang W, Hu W, Jiang W (2012) Comparison of five cisplatin-based regimens frequently used as the first-line protocols in metastatic nasopharyngeal carcinoma. J Cancer Res Clin Oncol 138:1717–1725. doi:10.1007/s00432-012-1219-x

    Article  CAS  PubMed  Google Scholar 

  • Kasai K, Ushio A, Kasai Y, Sawara K, Miyamoto Y, Suzuki K (2013) Therapeutic efficacy of transarterial chemo-embolization with a fine-powder formulation of cisplatin for hepatocellular carcinoma. World J Gastroenterol 19:2242–2248. doi:10.3748/wjg.v19.i14.2242

    Article  PubMed Central  PubMed  Google Scholar 

  • Lee EL, Shimizu T, Ise T, Numata T, Kohno K, Okada Y (2007) Impaired activity of volume-sensitive Cl channel is involved in cisplatin resistance of cancer cells. J Cell Physiol 211:513–521. doi:10.1002/jcp.20961

    Article  CAS  PubMed  Google Scholar 

  • Ma H, Zhou Z, Liang Y, Saxena S, Warnock DG (2004) Acidic ATP activates lymphocyte outwardly rectifying chloride channels via a novel pathway. Pflugers Arch 449:96–105. doi:10.1007/s00424-004-1305-2

    Article  CAS  PubMed  Google Scholar 

  • Mao J, Wang L, Sun X, Zhu L, Li P, Zhong P, Nie S, Jacob TJ, Chen L (2004) Volume-activated Cl current in migrated nasopharyngeal carcinoma cells. Sheng Li Xue Bao 56:525–530

    CAS  PubMed  Google Scholar 

  • Mao J, Wang L, Fan A, Wang J, Xu B, Jacob TJ, Chen L (2007) Blockage of volume-activated chloride channels inhibits migration of nasopharyngeal carcinoma cells. Cell Physiol Biochem 19:249–258

    Article  CAS  PubMed  Google Scholar 

  • Mao J, Chen L, Xu B, Wang L, Li H, Guo J, Li W, Nie S, Jacob TJ (2008) Suppression of ClC-3 channel expression reduces migration of nasopharyngeal carcinoma cells. Biochem Pharmacol 75:1706–1716. doi:10.1016/j.bcp.2008.01.008

    Article  CAS  PubMed  Google Scholar 

  • Mao J, Yuan J, Wang L, Zhang H, Jin X, Zhu J, Li H, Xu B, Chen L (2013) Tamoxifen inhibits migration of estrogen receptor-negative hepatocellular carcinoma cells by blocking the swelling-activated chloride current. J Cell Physiol 228:991–1001. doi:10.1002/jcp.24245

    Article  CAS  PubMed  Google Scholar 

  • Mehta N, Kaur M, Singh M, Chand S, Vyas B, Silakari P, Bahia MS, Silakari O (2014) Purinergic receptor P2X(7): a novel target for anti-inflammatory therapy. Bioorg Med Chem 22:54–88. doi:10.1016/j.bmc.2013.10.054

    Article  CAS  PubMed  Google Scholar 

  • Min X, Li H, Hou S, He W, Liu J, Hu B, Wang J (2011) Dysfunction of volume-sensitive chloride channels contributes to cisplatin resistance in human lung adenocarcinoma cells. Exp Biol Med (Maywood) 236:483–491. doi:10.1258/ebm.2011.010297

    Article  CAS  Google Scholar 

  • Moheimani F, Jackson DE (2012) P2Y12 receptor: platelet thrombus formation and medical interventions. Int J Hematol 96:572–587. doi:10.1007/s12185-012-1188-5

    Article  PubMed  Google Scholar 

  • Okada Y, Maeno E, Shimizu T, Dezaki K, Wang J, Morishima S (2001) Receptor-mediated control of regulatory volume decrease (RVD) and apoptotic volume decrease (AVD). J Physiol 532:3–16

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Roman RM, Wang Y, Lidofsky SD, Feranchak AP, Lomri N, Scharschmidt BF, Fitz JG (1997) Hepatocellular ATP-binding cassette protein expression enhances ATP release and autocrine regulation of cell volume. J Biol Chem 272:21970–21976

    Article  CAS  PubMed  Google Scholar 

  • Roman RM, Feranchak AP, Salter KD, Wang Y, Fitz JG (1999) Endogenous ATP release regulates Cl secretion in cultured human and rat biliary epithelial cells. Am J Physiol 276:G1391–G1400

    CAS  PubMed  Google Scholar 

  • Shen DW, Pouliot LM, Hall MD, Gottesman MM (2012) Cisplatin resistance: a cellular self-defense mechanism resulting from multiple epigenetic and genetic changes. Pharmacol Rev 64:706–721. doi:10.1124/pr.111.005637

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Shimizu T, Lee EL, Ise T, Okada Y (2008) Volume-sensitive Cl channel as a regulator of acquired cisplatin resistance. Anticancer Res 28:75–83

    CAS  PubMed  Google Scholar 

  • Sun X, Chen L, Luo H, Mao J, Zhu L, Nie S, Wang L (2012) Volume-activated chloride currents in fetal human nasopharyngeal epithelial cells. J Membr Biol 245:107–115. doi:10.1007/s00232-012-9419-5

    Article  CAS  PubMed  Google Scholar 

  • Tan J, You Y, Xu T, Yu P, Wu D, Deng H, Zhang Y, Bie P (2014) Par-4 downregulation confers cisplatin resistance in pancreatic cancer cells via PI3K/Akt pathway-dependent EMT. Toxicol Lett 224:7–15. doi:10.1016/j.toxlet.2013.10.008

    Article  CAS  PubMed  Google Scholar 

  • Varela D, Penna A, Simon F, Eguiguren AL, Leiva-Salcedo E, Cerda O, Sala F, Stutzin A (2010) P2X4 activation modulates volume-sensitive outwardly rectifying chloride channels in rat hepatoma cells. J Biol Chem 285:7566–7574. doi:10.1074/jbc.M109.063693

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Wang Y, Roman R, Lidofsky SD, Fitz JG (1996) Autocrine signaling through ATP release represents a novel mechanism for cell volume regulation. Proc Natl Acad Sci USA 93:12020–12025

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Xiao X, Yu S, Li S, Wu J, Ma R, Cao H, Zhu Y, Feng J (2014) Exosomes: decreased sensitivity of lung cancer A549 cells to Cisplatin. PLoS ONE 9:e89534. doi:10.1371/journal.pone.0089534

    Article  PubMed Central  PubMed  Google Scholar 

  • Yang L, Ye D, Ye W, Jiao C, Zhu L, Mao J, Jacob TJ, Wang L, Chen L (2011) ClC-3 is a main component of background chloride channels activated under isotonic conditions by autocrine ATP in nasopharyngeal carcinoma cells. J Cell Physiol 226:2516–2526. doi:10.1002/jcp.22596

    Article  CAS  PubMed  Google Scholar 

  • Zhang P, Liu H, Xia F, Zhang Q, Zhang Y, Zhao Q, Chao Z, Jiang Z, Jiang C (2014) Epithelial-mesenchymal transition is necessary for acquired resistance to cisplatin and increases the metastatic potential of nasopharyngeal carcinoma cells. Int J Mol Med 33:151–159. doi:10.3892/ijmm.2013.1538

    PubMed  Google Scholar 

  • Zhou J, Han J, Chen S, Xu Y, Kong Y, Xiao B, Tao Z (2012) gamma-Secretase inhibition combined with cisplatin enhances apoptosis of nasopharyngeal carcinoma cells. Exp Ther Med 3:357–361. doi:10.3892/etm.2011.410

    PubMed Central  CAS  PubMed  Google Scholar 

  • Zuo W, Zhu L, Bai Z, Zhang H, Mao J, Chen L, Wang L (2009) Chloride channels involve in hydrogen peroxide-induced apoptosis of PC12 cells. Biochem Biophys Res Commun 387:666–670. doi:10.1016/j.bbrc.2009.07.075

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (81173064, 81272223, 81273539), the Natural Science Foundation (S2011010001589) and the Medical Research Foundation (B2012187) of Guangdong Province of China, the Ministry of Education of China (20124401110009), and the Science and Technology Programs in Guangzhou (2013J500015) and Dongguan (2011108102006) of China.

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Correspondence to Lixin Chen or Liwei Wang.

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Yang, X., Zhu, L., Lin, J. et al. Cisplatin Activates Volume-Sensitive Like Chloride Channels Via Purinergic Receptor Pathways in Nasopharyngeal Carcinoma Cells. J Membrane Biol 248, 19–29 (2015). https://doi.org/10.1007/s00232-014-9724-2

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  • DOI: https://doi.org/10.1007/s00232-014-9724-2

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