Paroxetine-Induced Ca(superscript 2+) Movement and Death in OC2 Human Oral Cancer Cells

10.4077/CJP.2011.AMM046

Yi-Chien Fang；Chiang-Ting Chou；Chih-Chuan Pan；Yao-Dung Hsieh；Wei-Zhe Liang；David Chao；Jeng-Yu Tsai；Wei-Chuan Liao；Daih-Huang Kuo；Pochuen Shieh；Chun-Chi Kuo；Chung-Ren Jan；Chen-Fu Shaw

Ca(superscript 2+) ； OC2 ； oral cancer ； paroxetine

The Chinese Journal of Physiology

54卷5期（2011 / 10 / 01）

8 - 15

英文

The effect of the antidepressant paroxetine on cytosolic free Ca(superscript 2+) concentrations ([Ca(superscript 2+)](subscript i)) in OC2 human oral cancer cells is unclear. This study explored whether paroxetine changed basal [Ca(superscript 2+)](subscript i) levels in suspended OC2 cells by using fura-2 as a Ca(superscript 2+)-sensitive fluorescent dye. Paroxetine at concentrations between 100-1,000 μM increased [Ca(superscript 2+)](subscript i) in a concentration-dependent manner. The Ca(superscript 2+) signal was reduced by 50% by removing extracellular Ca(superscript 2+). Paroxetine-induced Ca(superscript 2+) influx was inhibited by the store-operated Ca(superscript 2+) channel blockers nifedipine, econazole and SK&F96365, and protein kinase C modulators. In Ca(superscript 2+)-free medium, pretreatment with the endoplasmic reticulum Ca(superscript 2+) pump inhibitor thapsigargin abolished paroxetine-induced [Ca(superscript 2+)](subscript i) rise. Inhibition of phospholipase C with U73122 did not alter paroxetine-induced [Ca(superscript 2+)](subscript i) rise. Paroxetine at 10-50 μM induced cell death in a concentrationdependent manner. The death was not reversed when cytosolic Ca(superscript 2+) was chelated with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Propidium iodide staining suggests that apoptosis plays a role in the death. Collectively, in OC2 cells, paroxetine induced [Ca(superscript 2+)](subscript i) rise by causing phospholipase C-independent Ca(superscript 2+) release from the endoplasmic reticulum and Ca(superscript 2+) influx via storeoperated Ca(superscript 2+) channels in a manner regulated by protein kinase C and phospholipase A2. Paroxetine (up to 50 μM) induced cell death in a Ca(superscript 2+)-independent manner.

1. Albert, A.P.,Large, W.A.(2002).Activation of store-operated channels by noradrenaline via protein kinase C in rabbit portal vein myocytes.J. Physiol.,544,113-125.
2. Bertini, F.,Costa, N.C.,Brandão, A.A.,Cavalcante, A.S.,Almeida, J.D.(2009).Ulceration of the oral mucosa induced by antidepressant medication: a case report.J. Med. Case Reports.,3,98.
3. Bootman, M.D.,Berridge, M.J.,Roderick, H.L.(2002).Calcium signalling: more messengers, more channels, more complexity.Cur. Biol.,12,R563-R565.
4. Chen, W.C.,Huang, C.C.,Huang, C.J.,Chien, J.M.,Lin, K.L.,Lu, Y.C.,Chen, I.S.,Liu, S.I.,Hsu, S.S.,Chang, H.T.,Chou, C.T.,Jan, C.R.(2008).Mechanism of paroxetine-induced cell death in renal tubular cells.Basic Clin. Pharmacol. Toxicol.,103,407-413.
5. Chou, C.T.,He, S.,Jan, C.R.(2007).Paroxetine-induced apoptosis in human osteosarcoma cells: activation of p38 MAP kinase and caspase-3 pathways without involvement of [Ca2+]i elevation.Toxicol. Appl. Pharmacol.,218,265-273.
6. Chu, S.T.,Huang, C.C.,Huang, C.J.,Cheng, J.S.,Chai, K.L.,Cheng, H.H.,Fang, Y.C.,Chi, C.C.,Su, H.H.,Chou, C.T.,Jan, C.R.(2007).Tamoxifen-induced [Ca2+]i rises and Ca2+-independent cell death in human oral cancer cells.J. Recept. Signal. Transduct. Res.,27,353-367.
7. Clapham, D.E.(1995).Intracellular calcium. Replenishing the stores.Nature,375,634-635.
8. Das, S.,Bhattacharyya, S.,Ghosh, S.,Majumdar, S.(1999).TNF-alpha induced altered signaling mechanism in human neutrophil.Mol. Cell. Biochem.,197,97-108.
9. Dick, I.E.,Brochu, R.M.,Purohit, Y.,Kaczorowski, G.J.,Martin, W.J.,Priest, B.T.(2007).Sodium channel blockade may contribute to the analgesic efficacy of antidepressants.J. Pain,8,315-324.
10. Gil-Ad, I.,Zolokov, A.,Lomnitski, L.,Taler, M.,Bar, M.,Luria, D.,Ram, E.,Weizman, A.(2008).Evaluation of the potential anti-cancer activity of the antidepressant sertraline in human colon cancer cell lines and in colorectal cancer-xenografted mice.Int. J. Oncol.,33,277-286.
11. Grynkiewicz, G.,Poenie, M.,Tsien, R.Y.(1985).A new generation of Ca2+ indicators with greatly improved fluorescence properties.J. Biol. Chem.,260,3440-3450.
12. Huang, C.,Huang, C.,Cheng, J.,Liu, S.,Chen, I.,Tsai, J.,Chou, C.,Tseng, P.,Jan, C.(2009).Fendiline-evoked [Ca2+]i rises and non-Ca2+- triggered cell death in human oral cancer cells.Hum. Exp. Toxicol.,28,41-48.
13. Huang, J.K.,Huang, C.J.,Chen, W.C.,Liu, S.I.,Hsu, S.S.,Chang, H.T.,Tseng, L.L.,Chou, C.T.,Chang, C.H.,Jan, C.R.(2005).Independent Ca2+ increases and cell proliferation induced by the carcinogen safrole in human oral cancer cells.Naunyn Schmiedebergs Arch. Pharmacol.,372,88-94.
14. Ishikawa, J.,Ohga, K.,Yoshino, T.,Takezawa, R.,Ichikawa, A.,Kubota, H.,Yamada, T.(2003).A pyrazole derivative, YM-58483, potently inhibits store-operated sustained Ca2+ influx and IL-2 production in T lymphocytes.J. Immunol.,170,4441-4449.
15. Jiang, N.,Zhang, Z.M.,Liu, L.,Zhang, C.,Zhang, Y.L.,Zhang, Z.C.(2006).Effects of Ca2+ channel blockers on store-operated Ca2+ channel currents of Kupffer cells after hepatic ischemia/reperfusion injury in rats.World J. Gastroenterol.,12,4694-4698.
16. Knorr, U.,Kessing, L.V.(2010).The effect of selective serotonin reuptake inhibitors in healthy subjects. A systematic review.Nord. J. Psychiatry,Jan
17. Kobayashi, T.,Washiyama, K.,Ikeda, K.(2006).Inhibition of G protein-activated inwardly rectifying K+ channels by the antidepressant paroxetine.J. Pharmacol. Sci.,102,278-287.
18. Korchak, H.M.,Corkey, B.E.,Yaney, G.C.,Kilpatrick, L.E.(2001).Negative regulation of ligand-initiated Ca2+ uptake by PKC-beta II in differentiated HL60 cells.Am. J. Physiol. Cell. Physiol.,281,C514-C523.
19. Kuo, L.N.,Huang, C.J.,Fang, Y.C.,Huang, C.C.,Wang, J.L.,Lin, K.L.,Chu, S.T.,Chang, H.T.,Chien, J.M.,Su, H.H.,Chi, C.C.,Chen, W.C.,Tsai, J.Y.,Liao, W.C.,Tseng, L.L.,Jan, C.R.(2009).Effect of thimerosal on Ca2+ movement and viability in human oral cancer cells.Hum. Exp. Toxicol.,28,301-308.
20. Large, W.A.,Saleh, S.N.,Albert, A.P.(2009).Role of phosphoinositol 4,5-bisphosphate and diacylglycerol in regulating native TRPC channel proteins in vascular smooth muscle.Cell Calcium,45,574-582.
21. Levkovitz, Y.,Gil-Ad, I.,Zeldich, E.,Dayag, M.,Weizman, A.(2005).Differential induction of apoptosis by antidepressants in glioma and neuroblastoma cell lines: evidence for p-c-Jun, cytochrome c, and caspase-3 involvement.J. Mol. Neurosci.,27,29-42.
22. Lin, C.C.,Yang, J.S.,Chen, J.T.,Fan, S.,Yu, F.S.,Yang, J.L.,Lu, C.C.,Kao, M.C.,Huang, A.C.,Lu, H.F.,Chung, J.G.(2007).Berberine induces apoptosis in human HSC-3 oral cancer cells via simultaneous activation of the death receptor-mediated and mitochondrial pathway.Anticancer Res.,27,3371-3378.
23. Ma, R.,Pluznick, J.,Kudlacek, P.,Sansom, S.C.(2001).Protein kinase C activates store-operated Ca2+ channels in human glomerular mesangial cells.J. Biol. Chem.,276,25759-25765.
24. Matuszyk, J.,Kobzdej, M.,Ziolo, E.,Kalas, W.,Kisielow, P.,Strzadala, L.(1998).Thymic lymphomas are resistant to Nur77-mediated apoptosis.Biochem. Biophys. Res. Commun.,249,279-282.
25. Merritt, J.E.,Jacob, R.,Hallam, T.J.(1989).Use of manganese to discriminate between calcium influx and mobilization from internal stores in stimulated human neutrophils.J. Biol. Chem.,264,1522-1527.
26. Nagata, K.,Imai, T.,Yamashita, T.,Tsuda, M.,Tozaki-Saitoh, H.,Inoue, K.(2009).Antidepressants inhibit P2X4 receptor function: a possible involvement in neuropathic pain relief.Mol. Pain,5,20.
27. Putney, J.W. Jr.(1986).A model for receptor-regulated calcium entry.Cell Calcium,7,1-12.
28. Quinn, T.,Molloy, M.,Smyth, A.,Baird, A.W.(2004).Capacitative calcium entry in guinea pig gallbladder smooth muscle in vitro.Life Sci.,74,1659-1669.
29. Rosetti, M.,Frasnelli, M.,Tesei, A.,Zoli, W.,Conti, M.(2006).Cytotoxicity of different selective serotonin reuptake inhibitors (SSRIs) against cancer cells.J. Exp. Ther. Oncol.,6,23-29.
30. Rundegren, J.,van Dijken, J.,Mörnstad, H.,von Knorring, L.(1985).Oral conditions in patients receiving long-term treatment with cyclic antidepressant drugs.Swed. Dent. J.,9,55-64.
31. Saleh, S.N.,Albert, A.P.,Peppiatt-Wildman, C.M.,Large, W.A.(2008).Diverse properties of store-operated TRPC channels activated by protein kinase C in vascular myocytes.J. Physiol.,586,2463-2476.
32. Schuster, C.,Fernbach, N.,Rix, U.,Superti-Furga, G.,Holy, M.,Freissmuth, M.,Sitte, H.H.,Sexl, V.(2007).Selective serotonin reuptake inhibitors-a new modality for the treatment of lymphoma/ leukaemia?.Biochem. Pharmacol.,74,1424-1435.
33. Shideman, C.R.,Reinardy, J.L.,Thayer, S.A.(2009).gamma-Secretase activity modulates store-operated Ca2+ entry into rat sensory neurons.Neurosci. Lett.,451,124-128.
34. Thastrup, O.,Cullen, P.J.,Drobak, B.K.,Hanley, M.R.,Dawson, A.P.(1990).Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2+-ATPase.Pro. Natl. Acad. Sci. U.S.A.,87,2466-2470.
35. Thompson, A.K.,Mostafapour, S.P.,Denlinger, L.C.,Bleasdale, J.E.,Fisher, S.K.(1991).The aminosteroid U-73122 inhibits muscarinic receptor sequestration and phosphoinositide hydrolysis in SK-N-SH neuroblastoma cells. A role for Gp in receptor compartmentation.J. Biol. Chem.,266,2385-2386.
36. Tsien, R.Y.(1980).New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures.Biochemistry,19,2396-2404.
37. Velez, L.I.,Shepherd, G.,Roth, B.A.,Benitez, F.L.(2003).Serotonin syndrome with elevated paroxetine concentrations.Ann. Pharmacotherapy,38,269-272.
38. Wagstaff, A.J.,Cheer, S.M.,Matheson, A.J.,Ormrod, D.,Goa, K.L.(2002).Paroxetine: an update of its use in psychiatric disorders in adults.Drugs,62,655-703.
39. Wang, G.K.,Mitchell, J.,Wang, S.Y.(2008).Block of persistent late Na+ currents by antidepressant sertraline and paroxetine.J. Membr. Biol.,222,79-90.

1. Liu, Shiuh-Inn,Liang, Wei-Zhe,Lai, Zhen-Rung,Jan, Chung-Ren,Huang, Jong-Khing,Chou, Chiang-Ting,Liu, Shiuh-Inn,Liang, Wei-Zhe,Lai, Zhen-Rung,Jan, Chung-Ren,Huang, Jong-Khing,Chou, Chiang-Ting(2013).Effect of BayK 8644 on [Ca2+] i and Viability in PC3 Human Prostate Cancer Cells.中國生理學雜誌,56(6),318-325.
2. Liu, Szu-Yuan,Liao, Wei-Chuan,Liang, Wei-Zhe,Kuo, Chun-Chi,Jan, Chung-Ren,Hsu, Shu-Shong,Chou, Chiang-Ting,Chen, Yu-Ting,Chen, Hsuan(2012).Deltamethrin-Induced [Ca2+]i Rise and Death in HGB Human Glioblastoma Cells.中國生理學雜誌,55(4),294-304.
3. Shieh, Pochuen,Liang, Wei-Zhe,Kuo, Daih-Huang,Kuo, Chun-Chi,Jan, Chung-Ren,Ho, Chin-Man,Chou, Chiang-Ting,Chien, Jau-Min(2014).The Mechanism of NPC-14686-Induced [Ca2+]i Rises and Non-Ca2+-Triggered Cell Death in MG63 Human Osteosarcoma Cells.中國生理學雜誌,57(3),158-166.
4. Sun, Te-Kung,Shieh, Pochuen,Lin, Jia-Rong,Liang, Wei-Zhe,Li, Yih-Do,Kuo, Daih-Huang,Kuo, Chun-Chi,Jan, Chung-Ren,Hung, Tzu-Yi,Ho, Chin-Man,Fang, Yi-Chien,Chou, Chiang-Ting,Cheng, Jin-Shiung(2015).The Mechanism of Safrole-Induced [Ca2+]i Rises and Non-Ca2+-Triggered Cell Death in SCM1 Human Gastric Cancer Cells.中國生理學雜誌,58(5),302-311.
5. Yeh, Jeng-Hsien,Yeh, Hsiao-Chun,Sun, Te-Kung,Shieh, Pochuen,Liang, Wei-Zhe,Lee, Jenn-Kuen,Kuo, Daih-Huang,Kuo, Chun-Chi,Jan, Chung-Ren,Chou, Chiang-Ting,Chen, Wei-Chuan(2015).Effect of Sertraline on Ca2+ Fluxes in Rabbit Corneal Epithelial Cells.中國生理學雜誌,58(2),85-94.
6. Yeh, Tzung Lieh,Yang, Yen Kuang,Wang, Yu-Shan,Lu, Ru-Band,Lee, I Hui,Chuang, Hui-Yu,Chu, Chun-Hsien,Cheng, Ling-Yi,Chen, Shiou-Lan,Chen, Shih-Heng,Chen, Po See,Chang, Yun-Hsuan(2014).Venlafaxine, Paroxetineand and Milnacipran for Major Depressive Disorder: A Pragmatic 24-Week Study.中國生理學雜誌,57(5),265-270.