Establishment of a new canine inflammatory mammary carcinoma cell line and analysis of its cystine-glutamate transporter subunit expression

Itoh, Harumichi; Naruse, Ryo; Tani, Kenji; Sunahara, Hiroshi; Nemoto, Yuki; Nakaichi, Munekazu; Iseri, Toshie; Horikirizono, Hiro; Itamoto, Kazuhito

doi:10.2478/jvetres-2022-0023

Abstract

Introduction

Inflammatory mammary carcinoma (IMC) is a rare disease with a poor prognosis and one affecting dogs. Inflammatory breast carcinoma (IBC) is a subtype of malignant breast cancer in humans with a high degree of malignancy and a similarly poor prognosis. Since the clinical symptoms and prognoses of both are similar, canine IMC has been considered as a model of human IBC. In this study, we newly established a stable IMC-derived cell line from a patient at the Yamaguchi University Animal Medical Center in Japan.

Material and Methods

The patient was a female toy poodle presenting with an inflamed mammary gland, which was diagnosed as IMC. The cell line was established from a tissue biopsy. Surface antigen marker (CD24 and CD44) expression was determined. Cystine/glutamate antiporter (xCT) expression was determined by Western blotting, flow cytometry and fluorescence immunostaining, and sulfasalazine was administered to ascertain if it suppressed xCT expression. Stem cell marker (Nanog, Sox2, Myc and Klf4) expression and aldehyde dehydrogenase (ALDH) activity were also investigated.

Results

The cultured cells showed xCT, and its suppression showed downregulation of stem cell markers and ALDH activity. Stable cell proliferation was verified.

Conclusion

A new canine IMC-derived cell line was established. In the future, we aim to study the effect of xCT on the maintenance of cancer stem cell properties in canine tumours, and propose a new therapeutic method for the treatment of canine IMC by targeting xCT.

References

Bonnet D., Dick J.E.: Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med 1997, 3, 730–737, doi: 10.1038/nm0797-730.
Open DOI Search in Google Scholar Back to article
Boughey J.C., Gonzalez R.J., Bonner E., Kuerer H.M.: Current treatment and clinical trial developments for ductal carcinoma in situ of the breast. Oncologist 2007, 12, 1276–1287, doi: 10.1634/theoncologist.12-11-1276.
Open DOI Search in Google Scholar Back to article
Caceres S., Pena L., de Andres P.J., Illera M.J., Lopez M.S., Woodward W.A., Reuben J.M., Illera J.C.: Establishment and characterization of a new cell line of canine inflammatory mammary cancer: IPC-366. PLoS One 2015, 10, e0122277, doi: 10.1371/journal.pone.0122277.
Open DOI Search in Google Scholar Back to article
Caceres S., Pena L., Lacerda L., Illera M.J., de Andres P.J., Larson R.A., Gao H., Debeb B.G., Woodward W.A., Reuben J.M., Illera J.C.: Canine cell line, IPC-366, as a good model for the study of inflammatory breast cancer. Vet Comp Oncol 2017, 15, 980–995, doi: 10.1111/vco.12238.
Open DOI Search in Google Scholar Back to article
de Mello Souza C.H., Toledo-Piza E., Amorin R., Barboza A., Tobias K.M.: Inflammatory mammary carcinoma in 12 dogs: clinical features, cyclooxygenase-2 expression, and response to piroxicam treatment. Can Vet J 2009, 50, 506–510.
Search in Google Scholar Back to article
Ginestier C., Hur M.H., Charafe-Jauffret E., Monville F., Dutcher J., Brown M., Jacquemier J., Viens P., Kleer C.G., Liu S., Schott A., Hayes D., Birnbaum D., Wicha M.S., Dontu G.: ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell 2007, 1, 555–567, doi: 10.1016/j.stem.2007.08.014.
Open DOI Search in Google Scholar Back to article
Hance K.W., Anderson W.F., Devesa S.S., Young H.A., Levine P.H.: Trends in inflammatory breast carcinoma incidence and survival: the surveillance, epidemiology, and end results program at the National Cancer Institute. J Natl Cancer Inst 2005, 97, 966–975, doi: 10.1093/jnci/dji172.
Open DOI Search in Google Scholar Back to article
Ishimoto T., Nagano O., Yae T., Tamada M., Motohara T., Oshima H., Oshima M., Ikeda T., Asaba R., Yagi H., Masuko T., Shimizu T., Ishikawa T., Kai K., Takahashi E., Imamura Y., Baba Y., Ohmura M., Suematsu M., Baba H., Saya H.: CD44 variant regulates redox status in cancer cells by stabilizing the xCT subunit of system xc(-) and thereby promotes tumor growth. Cancer Cell 2011, 19, 387–400, doi: 10.1016/j.ccr.2011.01.038.
Open DOI Search in Google Scholar Back to article
Jergens A.E., Schreiner C.A., Frank D.E., Niyo Y., Ahrens F.E., Eckersall P.D., Benson T.J., Evans R.: A scoring index for disease activity in canine inflammatory bowel disease. J Vet Intern Med 2003, 17, 291–297, doi: 10.1111/j.1939-1676.2003.tb02450.x.
Open DOI Search in Google Scholar Back to article
Kurebayashi J., Yamamoto S., Otsuki T., Sonoo H.: Medroxyprogesterone acetate inhibits interleukin 6 secretion from KPL-4 human breast cancer cells both in vitro and in vivo: a possible mechanism of the anticachectic effect. Br J Cancer 1999, 79, 631–636, doi: 10.1038/sj.bjc.6690099.
Open DOI Search in Google Scholar Back to article
Marcato P., Dean C.A., Pan D., Araslanova R., Gillis M., Joshi M., Helyer L., Pan L., Leidal A., Gujar S., Giacomantonio C.A., Lee P.W.: Aldehyde dehydrogenase activity of breast cancer stem cells is primarily due to isoform ALDH1A3 and its expression is predictive of metastasis. Stem Cells 2011, 29, 32–45, doi: 10.1002/stem.563.
Open DOI Search in Google Scholar Back to article
Maruo T., Kanemaki N., Onda K., Sato R., Ichihara N., Ochiai H.: Canine amino acid transport system Xc(-): cDNA sequence, distribution and cystine transport activity in lens epithelial cells. J Vet Med Sci 2014, 76, 523–530, doi: 10.1292/jvms.13-0170.
Open DOI Search in Google Scholar Back to article
Michishita M., Akiyoshi R., Suemizu H., Nakagawa T., Sasaki N., Takemitsu H., Arai T., Takahashi K.: Aldehyde dehydrogenase activity in cancer stem cells from canine mammary carcinoma cell lines. Vet J 2012, 193, 508–513, doi: 10.1016/j.tvjl.2012.01.006.
Open DOI Search in Google Scholar Back to article
Michishita M., Akiyoshi R., Yoshimura H., Katsumoto T., Ichikawa H., Ohkusu-Tsukada K., Nakagawa T., Sasaki N., Takahashi K.: Characterization of spheres derived from canine mammary gland adenocarcinoma cell lines. Res Vet Sci 2011, 91, 254–260, doi: 10.1016/j.rvsc.2010.11.016.
Open DOI Search in Google Scholar Back to article
Moreb J.S.: Aldehyde dehydrogenase as a marker for stem cells. Curr Stem Cell Res Ther 2008, 3, 237–246, doi: 10.2174/ 157488808786734006.
Open DOI Search in Google Scholar Back to article
Mu Z., Klinowska T., Dong X., Foster E., Womack C., Fernandez S.V., Cristofanilli M.: AZD8931, an equipotent, reversible inhibitor of signaling by epidermal growth factor receptor (EGFR), HER2, and HER3: preclinical activity in HER2 non-amplified inflammatory breast cancer models. J Exp Clin Cancer Res 2014, 33, 47, doi: 10.1186/1756-9966-33-47.
Open DOI Search in Google Scholar Back to article
Nishikawa S., Konno M., Hamabe A., Hasegawa S., Kano Y., Ohta K., Fukusumi T., Sakai D., Kudo T., Haraguchi N., Satoh T., Takiguchi S., Mori M., Doki Y., Ishii H.: Aldehyde dehydrogenase high gastric cancer stem cells are resistant to chemotherapy. Int J Oncol 2013, 42, 1437–1442, doi: 10.3892/ijo.2013.1837.
Open DOI Search in Google Scholar Back to article
Okazaki S., Shintani S., Hirata Y., Suina K., Semba T., Yamasaki J., Umene K., Ishikawa M., Saya H., Nagano O.: Synthetic lethality of the ALDH3A1 inhibitor dyclonine and xCT inhibitors in glutathione deficiency-resistant cancer cells. Oncotarget 2018, 9, 33832–33843, doi: 10.18632/oncotarget.26112.
Open DOI Search in Google Scholar Back to article
Otsubo K., Nosaki K., Imamura C.K., Ogata H., Fujita A., Sakata S., Hirai F., Toyokawa G., Iwama E., Harada T., Seto T., Takenoyama M., Ozeki T., Mushiroda T., Inada M., Kishimoto J., Tsuchihashi K., Suina K., Nagano O., Saya H., Nakanishi Y., Okamoto I.: Phase I study of salazosulfapyridine in combination with cisplatin and pemetrexed for advanced non-small-cell lung cancer. Cancer Sci 2017, 108, 1843–1849, doi: 10.1111/cas.13309.
Open DOI Search in Google Scholar Back to article
Perez Alenza M.D., Tabanera E., Pena L.: Inflammatory mammary carcinoma in dogs: 33 cases (1995-1999). J Am Vet Med Assoc 2001, 219, 1110–1114, doi: 10.2460/javma.2001.219.1110.
Open DOI Search in Google Scholar Back to article
Reya T., Morrison S.J., Clarke M.F., Weissman I.L.: Stem cells, cancer, and cancer stem cells. Nature 2001, 414, 105–111, doi: 10.1038/35102167.
Open DOI Search in Google Scholar Back to article
Ruiu R., Rolih V., Bolli E., Barutello G., Riccardo F., Quaglino E., Merighi I.F., Pericle F., Donofrio G., Cavallo F., Conti L.: Fighting breast cancer stem cells through the immune-targeting of the xCT cystine-glutamate antiporter. Cancer Immunol Immunother 2019, 68, 131–141, doi: 10.1007/s00262-018-2185-1.
Open DOI Search in Google Scholar Back to article
Sharma M.K., Seidlitz E.P., Singh G.: Cancer cells release glutamate via the cystine/glutamate antiporter. Biochem Biophys Res Commun 2010, 391, 91–95, doi: 10.1016/j.bbrc.2009.10.168.
Open DOI Search in Google Scholar Back to article
Shirakawa K., Tsuda H., Heike Y., Kato K., Asada R., Inomata M., Sasaki H., Kasumi F., Yoshimoto M., Iwanaga T., Konishi F., Terada M., Wakasugi H.: Absence of endothelial cells, central necrosis, and fibrosis are associated with aggressive inflammatory breast cancer. Cancer Res 2001, 61, 445–451.
Search in Google Scholar Back to article
Sun S., Wang Z.: ALDH high adenoid cystic carcinoma cells display cancer stem cell properties and are responsible for mediating metastasis. Biochem Biophys Res Commun 2010, 396, 843–848, doi: 10.1016/j.bbrc.2010.04.170.
Open DOI Search in Google Scholar Back to article
Tanabe A., Kimura K., Tazawa H., Maruo T., Taguchi M., Sahara H.: Functional analysis of CD44 variants and xCT in canine tumours. Vet Med Sci 2021, 7, 577–585, doi: 10.1002/vms3.397.
Open DOI Search in Google Scholar Back to article
Timmerman L.A., Holton T., Yuneva M., Louie R.J., Padro M., Daemen A., Hu M., Chan D.A., Ethier S.P., van ’t Veer L.J., Polyak K., McCormick F., Gray J.W.: Glutamine sensitivity analysis identifies the xCT antiporter as a common triple-negative breast tumor therapeutic target. Cancer Cell 2013, 24, 450–465, doi: 10.1016/j.ccr.2013.08.020.
Open DOI Search in Google Scholar Back to article
Uyama R., Nakagawa T., Hong S.H., Mochizuki M., Nishimura R., Sasaki N.: Establishment of four pairs of canine mammary tumour cell lines derived from primary and metastatic origin and their E-cadherin expression. Vet Comp Oncol 2006, 4, 104–113, doi: 10.1111/j.1476-5810.2006.00098.x.
Open DOI Search in Google Scholar Back to article
Van den Eynden G.G., Van Laere S.J., Van der Auwera I., Merajver S.D., Van Marck E.A., van Dam P., Vermeulen P.B., Dirix L.Y., van Golen K.L.: Overexpression of caveolin-1 and -2 in cell lines and in human samples of inflammatory breast cancer. Breast Cancer Res Treat 2006, 95, 219–228, doi: 10.1007/s10549-005-9002-1.
Open DOI Search in Google Scholar Back to article
Vassalli G.: Aldehyde Dehydrogenases: Not Just Markers, but Functional Regulators of Stem Cells. Stem Cells Int 2019, 2019, 3904645, doi: 10.1155/2019/3904645.
Open DOI Search in Google Scholar Back to article
Visvader J.E., Stingl J.: Mammary stem cells and the differentiation hierarchy: current status and perspectives. Genes Dev 2014, 28, 1143–1158, doi: 10.1101/gad.242511.114.
Open DOI Search in Google Scholar Back to article
Xiao Y., Ye Y., Yearsley K., Jones S., Barsky S.H.: The lymphovascular embolus of inflammatory breast cancer expresses a stem cell-like phenotype. Am J Pathol 2008, 173, 561–574, doi: 10.2353/ajpath.2008.071214.
Open DOI Search in Google Scholar Back to article
Yae T., Tsuchihashi K., Ishimoto T., Motohara T., Yoshikawa M., Yoshida G.J., Wada T., Masuko T., Mogushi K., Tanaka H., Osawa T., Kanki Y., Minami T., Aburatani H., Ohmura M., Kubo A., Suematsu M., Takahashi K., Saya H., Nagano O.: Alternative splicing of CD44 mRNA by ESRP1 enhances lung colonization of metastatic cancer cell. Nat Commun 2012, 3, 883, doi: 10.1038/ncomms1892.
Open DOI Search in Google Scholar Back to article
Yamauchi H., Woodward W.A., Valero V., Alvarez R.H., Lucci A., Buchholz T.A., Iwamoto T., Krishnamurthy S., Yang W., Reuben J.M., Hortobagyi G.N., Ueno N.T.: Inflammatory breast cancer: what we know and what we need to learn. Oncologist 2012, 17, 891–899, doi: 10.1634/theoncologist.2012-0039.
Open DOI Search in Google Scholar Back to article
Zhou Y., Yang J., Chen C., Li Z., Chen Y., Zhang X., Wang L., Zhou J.: Polyphyllin -Induced Ferroptosis in MDA-MB-231 Triple-Negative Breast Cancer Cells can Be Protected Against by KLF4-Mediated Upregulation of xCT. Front Pharmacol 2021, 12, 670224, doi: 10.3389/fphar.2021.670224.
Open DOI Search in Google Scholar Back to article