In vivo dual targeting of the oncogenic Ether-à-go-go-1 potassium channel by calcitriol and astemizole results in enhanced antineoplastic effects in breast tumors

BMC Cancer

Janice García-Quiroz⁵  Rocío García-Becerra⁵  Nancy Santos-Martínez⁵  David Barrera⁵  David Ordaz-Rosado⁵  Euclides Avila⁵  Ali Halhali⁵  Octavio Villanueva¹  Maŕa J Ibarra-Sánchez²  José Esparza-López²  Armando Gamboa-Domínguez⁴  Javier Camacho³  Fernando Larrea⁵  Lorenza Díaz⁵ 

[1] Departamento de Investigación Experimental y Bioterio, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, México, DF, México

[2] Departamento de Bioquímica, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, México, DF, México

[3] Departamento de Farmacología, Centro de Investigación y de Estudios Avanzados del I.P.N., Av. Instituto Politécnico Nacional No. 2508, Gustavo A. Madero, México, DF 07360, México

[4] Departamento de Patología, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, México, DF, México

[5] Departamento de Biología de la Reproducción, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Vasco de Quiroga No. 15, Tlalpan, México, DF 14000, México

关键词: EAG1;    Ki-67;    Targeted therapy;    Vitamin D receptor;    Breast cancer;    Vitamin D;   

Others  :  1120995
DOI  :  10.1186/1471-2407-14-745

 received in 2014-06-25, accepted in 2014-09-29,  发布年份 2014

【 摘 要 】

Background

The oncogenic ether-à-go-go-1 potassium channel (EAG1) activity and expression are necessary for cell cycle progression and tumorigenesis. The active vitamin D metabolite, calcitriol, and astemizole, a promising antineoplastic drug, target EAG1 by inhibiting its expression and blocking ion currents, respectively. We have previously shown a synergistic antiproliferative effect of calcitriol and astemizole in breast cancer cells in vitro, but the effect of this dual therapy in vivo has not been studied.

Methods

In the present study, we explored the combined antineoplastic effect of both drugs in vivo using mice xenografted with the human breast cancer cell line T-47D and a primary breast cancer-derived cell culture (MBCDF). Tumor-bearing athymic female mice were treated with oral astemizole (50 mg/kg/day) and/or intraperitoneal injections of calcitriol (0.03 μg/g body weight twice a week) during 3 weeks. Tumor sizes were measured thrice weekly. For mechanistic insights, we studied EAG1 expression by qPCR and Western blot. The expression of Ki-67 and the relative tumor volume were used as indicators of therapeutic efficacy.

Results

Compared to untreated controls, astemizole and calcitriol significantly reduced, while the coadministration of both drugs further suppressed, tumor growth (P < 0.05). In addition, the combined therapy significantly downregulated tumoral EAG1 and Ki-67 expression.

Conclusions

The concomitant administration of calcitriol and astemizole inhibited tumor growth more efficiently than each drug alone, which may be explained by the blocking of EAG1. These results provide the bases for further studies aimed at testing EAG1-dual targeting in breast cancer tumors expressing both EAG1 and the vitamin D receptor.

【 授权许可】

   
2014 García-Quiroz et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin 2011, 61:69-90.
• [2]Davis SL, Eckhardt SG, Tentler JJ, Diamond JR: Triple-negative breast cancer: bridging the gap from cancer genomics to predictive biomarkers. Ther Adv Med Oncol 2014, 6:88-100.
• [3]Wang Z: Roles of K + channels in regulating tumour cell proliferation and apoptosis. Pflugers Arch 2004, 448:274-286.
• [4]Camacho J: Ether a go-go potassium channels and cancer. Cancer Lett 2006, 233:1-9.
• [5]Pardo LA, Stühmer W: The roles of K(+) channels in cancer. Nat Rev Cancer 2014, 14:39-48.
• [6]Pardo LA, Stühmer W: Eag1 as a cancer target. Expert Opin Ther Targets 2008, 12:837-843.
• [7]Pardo LA, Del Camino D, Sanchez A, Alves F, Bruggemann A, Beckh S, Stühmer W: Oncogenic potential of EAG K(+) channels. EMBO J 1999, 18:5540-5547.
• [8]Diaz L, Ceja-Ochoa I, Restrepo-Angulo I, Larrea F, Avila-Chavez E, Garcia-Becerra R, Borja-Cacho E, Barrera D, Ahumada E, Gariglio P, Alvarez-Rios E, Ocadiz-Delgado R, Garcia-Villa E, Hernández-Gallegos E, Camacho-Arroyo I, Morales A, Ordaz-Rosado D, García-Latorre E, Escamilla J, Sánchez-Peña LC, Saqui-Salces M, Gamboa-Domínguez A, Vera E, Uribe-Ramirez M, Murbartián J, Ortiz CS, Rivera-Guevara C, De Vizcaya-Ruiz A, Camacho J: Estrogens and human papilloma virus oncogenes regulate human ether-a-go-go-1 potassium channel expression. Cancer Res 2009, 69:3300-3307.
• [9]Garcia-Becerra R, Diaz L, Camacho J, Barrera D, Ordaz-Rosado D, Morales A, Ortiz CS, Avila E, Bargallo E, Arrecillas M, Halhali A, Larrea F: Calcitriol inhibits Ether-a go-go potassium channel expression and cell proliferation in human breast cancer cells. Exp Cell Res 2010, 316:433-442.
• [10]Ouadid-Ahidouch H, Ahidouch A: K + channel expression in human breast cancer cells: involvement in cell cycle regulation and carcinogenesis. J Membr Biol 2008, 221:1-6.
• [11]Ouadid-Ahidouch H, Le Bourhis X, Roudbaraki M, Toillon RA, Delcourt P, Prevarskaya N: Changes in the K + current-density of MCF-7 cells during progression through the cell cycle: possible involvement of a h-ether.a-gogo K + channel. Receptors Channels 2001, 7:345-356.
• [12]Ouadid-Ahidouch H, Roudbaraki M, Ahidouch A, Delcourt P, Prevarskaya N: Cell-cycle-dependent expression of the large Ca2 + -activated K + channels in breast cancer cells. Biochem Biophys Res Commun 2004, 316:244-251.
• [13]Avila E, Garcia-Becerra R, Rodriguez-Rasgado JA, Diaz L, Ordaz-Rosado D, Zugel U, Steinmeyer A, Barrera D, Halhali A, Larrea F, Camacho J: Calcitriol down-regulates human ether a go-go 1 potassium channel expression in cervical cancer cells. Anticancer Res 2010, 30:2667-2672.
• [14]Deeb KK, Trump DL, Johnson CS: Vitamin D signalling pathways in cancer: potential for anticancer therapeutics. Nat Rev Cancer 2007, 7:684-700.
• [15]Krishnan AV, Trump DL, Johnson CS, Feldman D: The role of vitamin D in cancer prevention and treatment. Endocrinol Metab Clin North Am 2010, 39:401-418. table of contents
• [16]Welsh J: Cellular and molecular effects of vitamin D on carcinogenesis. Arch Biochem Biophys 2012, 523:107-114.
• [17]Garcia-Quiroz J, Garcia-Becerra R, Barrera D, Santos N, Avila E, Ordaz-Rosado D, Rivas-Suarez M, Halhali A, Rodriguez P, Gamboa-Dominguez A, Medina-Franco H, Camacho J, Larrea F, Díaz L: Astemizole synergizes calcitriol antiproliferative activity by inhibiting CYP24A1 and upregulating VDR: a novel approach for breast cancer therapy. PLoS One 2012, 7:e45063.
• [18]Berger U, Wilson P, McClelland RA, Colston K, Haussler MR, Pike JW, Coombes RC: Immunocytochemical detection of 1,25-dihydroxyvitamin D receptors in normal human tissues. J Clin Endocrinol Metab 1988, 67:607-613.
• [19]Freake HC, Abeyasekera G, Iwasaki J, Marcocci C, MacIntyre I, McClelland RA, Skilton RA, Easton DF, Coombes RC: Measurement of 1,25-dihydroxyvitamin D3 receptors in breast cancer and their relationship to biochemical and clinical indices. Cancer Res 1984, 44:1677-1681.
• [20]Friedrich M, Axt-Fliedner R, Villena-Heinsen C, Tilgen W, Schmidt W, Reichrath J: Analysis of vitamin D-receptor (VDR) and retinoid X-receptor alpha in breast cancer. Histochem J 2002, 34:35-40.
• [21]Hemmerlein B, Weseloh RM, Mello De Queiroz F, Knotgen H, Sanchez A, Rubio ME, Martin S, Schliephacke T, Jenke M, Heinz-Joachim R, Stühmer W, Pardo LA: Overexpression of Eag1 potassium channels in clinical tumours. Mol Cancer 2006, 5:41. BioMed Central Full Text
• [22]Esparza-Lopez J, Medina-Franco H, Escobar-Arriaga E, Leon-Rodriguez E, Zentella-Dehesa A, Ibarra-Sanchez MJ: Doxorubicin induces atypical NF-kappaB activation through c-Abl kinase activity in breast cancer cells. J Cancer Res Clin Oncol 2013, 139:1625-1635.
• [23]Downie BR, Sanchez A, Knotgen H, Contreras-Jurado C, Gymnopoulos M, Weber C, Stühmer W, Pardo LA: Eag1 expression interferes with hypoxia homeostasis and induces angiogenesis in tumors. J Biol Chem 2008, 283:36234-36240.
• [24]Krishnan AV, Swami S, Peng L, Wang J, Moreno J, Feldman D: Tissue-selective regulation of aromatase expression by calcitriol: implications for breast cancer therapy. Endocrinology 2010, 151:32-42.
• [25]Jacoby R, Fox J: Biology and disease of mice. In Laboratory Animal Medicine. Edited by Fox LCA J, Loew FM, Quimby FW. San Diego, CA: Academic Press, Inc; 1984:31-89.
• [26]Napp J, Monje F, Stühmer W, Pardo LA: Glycosylation of Eag1 (Kv10.1) potassium channels: intracellular trafficking and functional consequences. J Biol Chem 2005, 280:29506-29512.
• [27]Pardo LA, Stühmer W: Eag1: an emerging oncological target. Cancer Res 2008, 68:1611-1613.
• [28]Matthews D, LaPorta E, Zinser GM, Narvaez CJ, Welsh J: Genomic vitamin D signaling in breast cancer: Insights from animal models and human cells. J Steroid Biochem Mol Biol 2010, 121:362-367.
• [29]Chiang KC, Chen TC: The anti-cancer actions of vitamin D. Anticancer Agents Med Chem 2013, 13:126-139.
• [30]Chen Y, Sanchez A, Rubio ME, Kohl T, Pardo LA, Stühmer W: Functional K(v)10.1 channels localize to the inner nuclear membrane. PLoS One 2011, 6:e19257.
• [31]Krishnan AV, Swami S, Feldman D: Equivalent anticancer activities of dietary vitamin D and calcitriol in an animal model of breast cancer: Importance of mammary CYP27B1 for treatment and prevention. J Steroid Biochem Mol Biol 2012, 136:289-295.
• [32]Garcia-Quiroz J, Camacho J: Astemizole: an old anti-histamine as a new promising anti-cancer drug. Anticancer Agents Med Chem 2011, 11:307-314.
• [33]Laduron PM, Janssen PF, Gommeren W, Leysen JE: In vitro and in vivo binding characteristics of a new long-acting histamine H1 antagonist, astemizole. Mol Pharmacol 1982, 21:294-300.