Roles of voltage‑gated potassium channels in the maintenance of pancreatic cancer stem cells

Shiozaki,Atsushi; Konishi,Tomoki; Kosuga,Toshiyuki; Kudou,Michihiro; Kurashima,Kento; Inoue,Hiroyuki; Shoda,Katsutoshi; Arita,Tomohiro; Konishi,Hirotaka; Morimura,Ryo; Komatsu,Shuhei; Ikoma,Hisashi; Toma,Atsushi; Kubota,Takeshi; Fujiwara,Hitoshi; Okamoto,Kazuma; Otsuji,Eigo

doi:10.3892/ijo.2021.5256

Roles of voltage‑gated potassium channels in the maintenance of pancreatic cancer stem cells

Authors:
- Atsushi Shiozaki
- Tomoki Konishi
- Toshiyuki Kosuga
- Michihiro Kudou
- Kento Kurashima
- Hiroyuki Inoue
- Katsutoshi Shoda
- Tomohiro Arita
- Hirotaka Konishi
- Ryo Morimura
- Shuhei Komatsu
- Hisashi Ikoma
- Atsushi Toma
- Takeshi Kubota
- Hitoshi Fujiwara
- Kazuma Okamoto
- Eigo Otsuji
View Affiliations

Affiliations: Division of Digestive Surgery, Department of Surgery, Kyoto Prefectural University of Medicine, Kyoto 602‑8566, Japan
Published online on: August 12, 2021 https://doi.org/10.3892/ijo.2021.5256
Article Number: 76
Copyright: © Shiozaki et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The targeting of membrane proteins that are activated in cancer stem cells (CSCs) represents one of the key recent strategies in cancer therapy. The present study analyzed ion channel expression profiles and functions in pancreatic CSCs (PCSCs). Cells strongly expressing aldehyde dehydrogenase 1 family member A1 (ALDH1A1) were isolated from the human pancreatic PK59 cell line using fluorescence‑activated cell sorting, and PCSCs were identified based on tumorsphere formation. Microarray analysis was performed to investigate the gene expression profiles in PCSCs. ALDH1A1 messenger RNA levels were higher in PCSCs compared with non‑PCSCs. PCSCs were resistant to 5‑fluorouracil and capable of redifferentiation. The results of the microarray analysis revealed that gene expression related to ion channels, including voltage‑gated potassium channels (Kv), was upregulated in PCSCs compared with non‑PCSCs. 4‑Aminopyridine (4‑AP), a potent Kv inhibitor, exhibited greater cytotoxicity in PCSCs compared with non‑PCSCs. In a xenograft model in nude mice, tumor volumes were significantly lower in mice inoculated with PK59 cells pre‑treated with 4‑AP compared with those in mice injected with non‑treated cells. The present results identified a role of Kv in the persistence of PCSCs and suggested that the Kv inhibitor 4‑AP may have potential as a therapeutic agent for pancreatic carcinoma.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2017. CA Cancer J Clin. 67:7–30. 2017. View Article : Google Scholar
2	Ferlay J, Steliarova-Foucher E, Lortet-Tieulent J, Rosso S, Coebergh JW, Comber H, Forman D and Bray F: Cancer incidence and mortality patterns in Europe Estimates for 40 countries in 2012. Eur J Cancer. 49:1374–1403. 2013. View Article : Google Scholar
3	Ilic M and Ilic I: Epidemiology of pancreatic cancer. World J Gastroenterol. 22:9694–9705. 2016. View Article : Google Scholar
4	Ishiwata T, Matsuda Y, Yoshimura H, Sasaki N, Ishiwata S, Ishikawa N, Takubo K, Arai T and Aida J: Pancreatic cancer stem cells: Features and detection methods. Pathol Oncol Res. 2:797–805. 2018. View Article : Google Scholar
5	Moore MJ, Goldstein D, Hamm J, Figer A, Hecht JR, Gallinger S, Au HJ, Murawa P, Walde D, Wolff RA, et al: Erlotinib plus gemcitabine compared with gemcitabine alone in patients with advanced pancreatic cancer: A phase III trial of the national cancer institute of Canada clinical trials group. J Clin Oncol. 25:1960–1966. 2007. View Article : Google Scholar
6	Louvet C, Labianca R, Hammel P, Lledo G, Zampino MG, André T, Zaniboni A, Ducreux M, Aitini E, Taïeb J, et al: Gemcitabine in combination with oxaliplatin compared with gemcitabine alone in locally advanced or metastatic pancreatic cancer: Results of a GERCOR and GISCAD phase III trial. J Clin Oncol. 23:3509–3516. 2005. View Article : Google Scholar
7	Hu G, Li F, Ouyang K, Xie F, Tang X, Wang K, Han S, Jiang Z, Zhu M, Wen D, et al: Intrinsic gemcitabine resistance in a novel pancreatic cancer cell line is associated with cancer stem cell-like phenotype. Int J Oncol. 40:798–806. 2012.
8	Clarke MF, Dick JE, Dirks PB, Eaves CJ, Jamieson CH, Jones DL, Visvader J, Weissman IL and Wahl GM: Cancer stem cells-perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res. 66:9339–9344. 2006. View Article : Google Scholar
9	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: Accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar
10	Toh TB, Lim JJ and Chow EK: Epigenetics in cancer stem cells. Mol Cancer. 16:292017. View Article : Google Scholar
11	Long A, Giroux V, Whelan K A, Hamilton KE, Tetreault MP, Tanaka K, Lee JS, Klein-Szanto AJ, Nakagawa H and Rustgi AK: WNT10A promotes an invasive and self-renewing phenotype in esophageal squamous cell carcinoma. Carcinogenesis. 36:598–606. 2015. View Article : Google Scholar
12	Chen Q, Song S, Wei S, Liu B, Honjo S, Scott A, Jin J, Ma L, Zhu H, Skinner HD, et al: ABT-263 induces apoptosis and synergizes with chemotherapy by targeting stemness pathways in esophageal cancer. Oncotarget. 6:25883–25896. 2015. View Article : Google Scholar
13	Yue D, Zhang Z, Li J, Chen X, Ping Y, Liu S, Shi X, Li L, Wang L, Huang L, et al: Transforming growth factor-betal promotes the migration and invasion of sphere-forming stem-like cell subpopulations in esophageal cancer. Exp Cell Res. 336:141–149. 2015. View Article : Google Scholar
14	Brungs D, Aghmesheh M, Vine KL, Becker TM, Carolan MG and Ranson M: Gastric cancer stem cells: Evidence, potential markers, and clinical implications. J Gastroenterol. 51:313–326. 2016. View Article : Google Scholar
15	Li Y, Rogoff HA, Keates S, Gao Y, Murikipudi S, Mikule K, Leggett D, Li W, Pardee AB and Li CJ: Suppression of cancer relapse and metastasis by inhibiting cancer stemness. Proc Natl Acad Sci USA. 112:1839–1844. 2015. View Article : Google Scholar
16	Shiozaki A, Ichikawa D, Otsuji E and Marunaka Y: Cellular physiological approach for treatment of gastric cancer. World J Gastroenterol. 20:11560–11566. 2014. View Article : Google Scholar
17	Shiozaki A, Ariyoshi Y, Iitaka D, Kosuga T, Shimizu H, Kudou M, Konishi T, Shoda K, Arita T, Konishi H, et al: Functional analysis and clinical significance of sodium iodide symporter expression in gastric cancer. Gastric Cancer. 22:473–485. 2019. View Article : Google Scholar
18	Kosuga T, Shiozaki A, Kudou M, Yamazato Y, Ichikawa D, Komatsu S, Konishi H, Okamoto K, Shoda K, Arita T, et al: Blockade of potassium ion transports enhances hypotonicity-induced cytocidal effects in gastric cancer. Oncotarget. 8:101394–101405. 2017. View Article : Google Scholar
19	Kudou M, Shiozaki A, Yamazato Y, Katsurahara K, Kosuga T, Shoda K, Arita T, Konishi H, Komatsu S, Kubota T, et al: The expression and role of TRPV2 in esophageal squamous cell carcinoma. Sci Rep. 9:160552019. View Article : Google Scholar
20	Shiozaki A, Kudou M, Ichikawa D, Fujiwara H, Shimizu H, Ishimoto T, Arita T, Kosuga T, Konishi H, Komatsu S, et al: Esophageal cancer stem cells are suppressed by tranilast, a TRPV2 channel inhibitor. J Gastroenterol. 53:197–207. 2018. View Article : Google Scholar
21	Darakhshan S and Pour AB: Tranilast: A review of its therapeutic applications. Pharmacol Res. 91:15–28. 2015. View Article : Google Scholar
22	Almanaa TN, Geusz ME and Jamasbi RJ: A new method for identifying stem-like cells in esophageal cancer cell lines. J Cancer. 4:536–548. 2013. View Article : Google Scholar
23	Johnson S, Chen H and Lo PK: In vitro tumorsphere formation assays. Bio Protoc. 3:e3252013. View Article : Google Scholar
24	Miyazaki H, Shiozaki A, Niisato N and Marunaka Y: Physiological significance of hypotonicity-induced regulatory volume decrease: Reduction in intracellular Cl⁻ concentration acting as an intracellular signaling. Am J Physiol Renal Physiol. 292:F1411–F1417. 2007. View Article : Google Scholar
25	Fan P, Zhang Y, Liu L, Zhao Z, Yin Y, Xiao X, Bauer N, Gladkich J, Mattern J, Gao C, et al: Continuous exposure of pancreatic cancer cells to dietary bioactive agents does not induce drug resistance unlike chemotherapy. Cell Death Dis. 7:e22462016. View Article : Google Scholar
26	Rasheed ZA and Matsui W: Biological and clinical relevance of stem cells in pancreatic adenocarcinoma. J Gastroenterol Hepatol. 27(Suppl 2): S15–S18. 2012. View Article : Google Scholar
27	Matsuda Y, Kure S and Ishiwata T: Nestin and other putative cancer stem cell markers in pancreatic cancer. Med Mol Morphol. 45:59–65. 2012. View Article : Google Scholar
28	Miyazaki H, Shiozaki A, Niisato N, Ohsawa R, Itoi H, Ueda Y, Otsuji E, Yamagishi H, Iwasaki Y, Nakano T, et al: Chloride ions control the G1/S cell-cycle checkpoint by regulating the expression of p21 through a p53-independent pathway in human gastric cancer cells. Biochem Biophys Res Commun. 366:506–512. 2008. View Article : Google Scholar
29	Ohsawa R, Miyazaki H, Niisato N, Shiozaki A, Iwasaki Y, Otsuji E and Marunaka Y: Intracellular chloride regulates cell proliferation through the activation of stress-activated protein kinases in MKN28 human gastric cancer cells. J Cell Physiol. 223:764–770. 2010.
30	Shiozaki A, Otsuji E and Marunaka Y: Intracellular chloride regulates the G(1)/S cell cycle progression in gastric cancer cells. World J Gastrointest Oncol. 3:119–122. 2011. View Article : Google Scholar
31	Tanaka S, Miyazaki H, Shiozaki A, Ichikawa D, Otsuji E and Marunaka Y: Cytosolic Cl⁻ affects the anticancer activity of paclitaxel in the gastric cancer cell line, MKN28 cell. Cell Physiol Biochem. 42:68–80. 2017. View Article : Google Scholar
32	Rausch V, Liu L, Kallifatidis G, Baumann B, Mattern J, Gladkich J, Wirth T, Schemmer P, Büchler MW, Zoller M, et al: Synergistic activity of sorafenib and sulforaphane abolishes pancreatic cancer stem cell characteristics. Cancer Res. 70:5004–5013. 2010. View Article : Google Scholar
33	Kim MP, Fleming JB, Wang H, Abbruzzese JL, Choi W, Kopetz S, McConkey DJ, Evans DB and Gallick GE: ALDH activity selectively defines an enhanced tumor-initiating cell population relative to CD133 expression in human pancreatic adenocarcinoma. PLoS One. 6:e206362011. View Article : Google Scholar
34	Rasheed ZA, Yang J, Wang Q, Kowalski J, Freed I, Murter C, Hong SM, Koorstra JB, Rajeshkumar NV, He X, et al: Prognostic significance of tumorigenic cells with mesenchymal features in pancreatic adenocarcinoma. J Natl Cancer Ins. 102:340–351. 2010. View Article : Google Scholar
35	Wu HY, Yang MC, Ding LY, Chen CS and Chu PC: p21-Activated kinase 3 promotes cancer stem cell phenotypes through activating the Akt-GSK3ß-ß-catenin signaling pathway in pancreatic cancer cells. Cancer Lett. 456:13–22. 2019. View Article : Google Scholar
36	Comes N, Serrano-Albarras A, Capera J, Serrano-Novillo C, Condom E, Ramon Y, Cajal S, Ferreres JC and Felipe A: Involvement of potassium channels in the progression of cancer to a more malignant phenotype. Biochim Biophys Acta. 1848:2477–2492. 2015. View Article : Google Scholar
37	Rao VR, Perez-Neut M, Kaja S and Gentile S: Voltage-gated ion channels in cancer cell proliferation. Cancers (Basel). 7:849–875. 2015. View Article : Google Scholar
38	Serrano-Novillo C, Capera J, Colomer-Molera M, Condom E, Ferreres JC and Felipe A: Implication of voltage-gated potassium channels in neoplastic cell proliferation. Cancers (Basel). 11:2872019. View Article : Google Scholar
39	Brevet M, Fucks D, Chatelain D, Regimbeau JM, Delcenserie R, Sevestre H and Ouadid-Ahidouch H: Deregulation of 2 potassium channels in pancreas adenocarcinomas: Implication of KV1.3 gene promoter methylation. Pancreas. 38:649–654. 2009. View Article : Google Scholar
40	Leanza L, Zoratti M, Gulbins E and Szabo I: Mitochondrial ion channels as oncological targets. Oncogene. 33:5569–5581. 2014. View Article : Google Scholar
41	Zaccagnino A, Manago A, Leanza L, Gontarewitz A, Linder B, Azzolini M, Biasutto L, Zoratti M, Peruzzo R, Legler K, et al: Tumor-reducing effect of the clinically used drug clofazimine in a SCID mouse model of pancreatic ductal adenocarcinoma. Oncotarget. 8:38276–38293. 2017. View Article : Google Scholar
42	Leanza L, Romio M, Becker KA, Azzolini M, Trentin L, Manago A, Venturini E, Zaccagnino A, Mattarei A, Carraretto L, et al: Direct pharmacological targeting of a mitochondrial ion channel selectively kills tumor cells in vivo. Cancer Cell. 31:516–531.e10. 2017. View Article : Google Scholar
43	Szabo I, Trentin L, Trimarco V, Semenzato G and Leanza L: Biophysical characterization and expression analysis of Kv1.3 potassium channel in primary human leukemic B cells. Cell Physiol Biochem. 37:965–978. 2015. View Article : Google Scholar
44	Aissaoui D, Mlayah-Bellalouna S, Jebali J, Abdelkafi-Koubaa Z, Souid S, Moslah W, Othman H, Luis J, ElAyeb M, Marrakchi N, et al: Functional role of Kv1.1 and Kv1.3 channels in the neoplastic progression steps of three cancer cell lines, elucidated by scorpion peptides. Int J Biol Macromol. 111:1146–1155. 2018. View Article : Google Scholar
45	Martinez R, Stühmer W, Martin S, Schell J, Reichmann A, Rohde V and Pardo L: Analysis of the expression of Kv10.1 potassium channel in patients with brain metastases and glioblastoma multiforme: Impact on survival. BMC Cancer. 15:8392015. View Article : Google Scholar
46	Sette A, Spadavecchia J, Landoulsi J, Casale S, Haye B, Crociani O and Arcangeli A: Development of novel anti-Kv 11.1 antibody-conjugated PEG-TiO₂ nanoparticles for targeting pancreatic ductal adenocarcinoma cells. J Nanopart Res. 15:21112013. View Article : Google Scholar
47	Crociani O, Lastraioli E, Boni L, Pillozzi S, Romoli MR, D'Amico M, Stefanini M, Crescioli S, Masi A, Taddei A, et al: hERG1 channels regulate VEGF-A secretion in human gastric cancer: Clinicopathological correlations and therapeutical implications. Clin Cancer Res. 20:1502–1512. 2014. View Article : Google Scholar
48	Wang SP, Wang JA, Luo RH, Cui WY and Wang H: Potassium channel currents in rat mesenchymal stem cells and their possible roles in cell proliferation. Clin Exp Pharmacol Physiol. 35:1077–1084. 2008. View Article : Google Scholar
49	Zhang YY, Yue J, Che H, Sun HY, Tse HF and Li GR: BKCa and hEag1 channels regulate cell proliferation and differentiation in human bone marrow-derived mesenchymal stem cells. J Cell Physiol. 229:202–212. 2014. View Article : Google Scholar
50	Morokuma J, Blackiston D, Adams DS, Seebohm G, Trimmer B and Levin M: Modulation of potassium channel function confers a hyperproliferative invasive phenotype on embryonic stem cells. Proc Natl Acad Sci USA. 105:16608–16613. 2008. View Article : Google Scholar
51	Bai X, Ma J, Pan Z, Song YH, Freyberg S, Yan Y, Vykoukal D and Alt E: Electrophysiological properties of human adipose tissue-derived stem cells. Am J Physiol Cell Physiol. 293:C1539–C1550. 2007. View Article : Google Scholar
52	Jensen HB, Ravnborg M, Dalgas U and Stenager E: 4-Aminopyridine for symptomatic treatment of multiple sclerosis: A systematic review. Ther Adv Neurol Disord. 7:97–113. 2014. View Article : Google Scholar
53	Solari A, Uitdehaag B, Giuliani G, Pucci E and Taus C: Aminopyridines for symptomatic treatment in multiple sclerosis. Cochrane Database Syst Rev. 2002:CD0013302002.
54	Goodman AD, Brown TR, Edwards KR, Krupp LB, Schapiro RT, Cohen R, Marinucci LN and Blight AR; MSF204 Investigators: A phase 3 trial of extended release oral dalfampridine in multiple sclerosis. Ann Neurol. 68:494–502. 2010. View Article : Google Scholar
55	Goodman AD, Brown TR, Krupp LB, Schapiro RT, Schwid SR, Cohen R, Marinucci LN and Blight AR; Fampridine MS-F203 Investigators: Sustained-release oral fampridine in multiple sclerosis: A randomised, double-blind, controlled trial. Lancet. 373:732–738. 2009. View Article : Google Scholar
56	Rouzaire-Dubois B, Gerard V and Dubois JM: Involvement of K+ channels in the quercetin-induced inhibition of neuroblastoma cell growth. Pflugers Arch. 423:202–205. 1993. View Article : Google Scholar
57	Fieber LA, Gonzâlez DM, Wallace MR and Muir D: Delayed rectifier K currents in NF1 Schwann cells. Pharmacological block inhibits proliferation. Neurobiol Dis. 13:136–146. 2003. View Article : Google Scholar
58	Nilius B and Wohlrab W: Potassium Channels and Regulation of proliferation of human melanoma cells. J Physiol. 445:537–548. 1992. View Article : Google Scholar
59	Rybalchenko V, Prevarskaya N, Van Coppenolle F, Legrand G, Lemonnier L, Le Bourhis X and Skryma R: Verapamil inhibits proliferation of LNCaP human prostate cancer cells influencing K+ channel gating. Mol Pharmacol. 59:1376–1387. 2001. View Article : Google Scholar
60	Chin LS, Park CC, Zitnay KM, Sinha M, DiPatri AJ Jr, Perillan P and Simard JM: 4-Aminopyridine causes apoptosis and blocks an outward rectifier K+ channel in malignant astrocytoma cell lines. J Neurosci Res. 48:122–127. 1997. View Article : Google Scholar
61	Kim JA, Kang YS, Jung MW, Kang GH, Lee SH and Lee YS: Ca2⁺ influx mediates apoptosis induced by 4-aminopyridine, a K+ channel blocker, in HepG2 human hepatoblastoma cells. Pharmacology. 60:74–81. 2000. View Article : Google Scholar
62	Wang W, Xiao J, Adachi M, Liu Z and Zhou J: 4-aminopyridine induces apoptosis of human acute myeloid leukemia cells via increasing [Ca2⁺]i through P2X7 receptor pathway. Cell Physiol Biochem. 28:199–208. 2011. View Article : Google Scholar
63	Huang L, Li B, Li W, Guo H and Zou F: ATP-sensitive potassium channels control glioma cells proliferation by regulating ERK activity. Carcinogenesis. 30:737–744. 2009. View Article : Google Scholar
64	Ru Q, Li WL, Xiong Q, Chen L, Tian X and Li CY: Voltage-gated potassium channel blocker 4-aminopyridine induces glioma cell apoptosis by reducing expression of microRNA-10b-5p. Mol Biol Cell. 29:1125–1136. 2018. View Article : Google Scholar
65	Luo Z, Wang J, Li C, Qiu Y, Huang J, Huang Y, Gu H, Wu B, Hu Z and Zhen Y: Upregulation of phosphatase and tensin homolog is essential for the effect of 4-aminopyridine on A549/CDDP cells. Mol Med Rep. 17:5996–6001. 2018.