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一种新的犬炎性乳腺癌细胞系的建立及其胱氨酸-谷氨酸转运体亚基表达分析。

Establishment of a New Canine Inflammatory Mammary Carcinoma Cell Line and Analysis of its Cystine-glutamate Transporter Subunit Expression.

作者信息

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

机构信息

Department of Small Animal Clinical Science, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.

Department of Veterinary Surgery, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.

出版信息

J Vet Res. 2022 May 31;66(2):273-279. doi: 10.2478/jvetres-2022-0023. eCollection 2022 Jun.

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.

摘要

引言

炎性乳腺癌(IMC)是一种预后不良的罕见疾病,且在犬类中也有发生。炎性乳腺癌(IBC)是人类恶性乳腺癌的一种亚型,恶性程度高,预后同样不良。由于两者的临床症状和预后相似,犬类IMC被视为人类IBC的模型。在本研究中,我们从日本山口大学动物医学中心的一名患者身上新建立了一种稳定的IMC来源的细胞系。

材料与方法

该患者为一只雌性玩具贵宾犬,乳腺发炎,被诊断为IMC。该细胞系从组织活检中建立。测定了表面抗原标志物(CD24和CD44)的表达。通过蛋白质印迹法、流式细胞术和荧光免疫染色测定胱氨酸/谷氨酸反向转运体(xCT)的表达,并给予柳氮磺胺吡啶以确定其是否抑制xCT表达。还研究了干细胞标志物(Nanog、Sox2、Myc和Klf4)的表达以及醛脱氢酶(ALDH)的活性。

结果

培养的细胞显示出xCT表达,其抑制显示干细胞标志物和ALDH活性下调。证实了细胞的稳定增殖。

结论

建立了一种新的犬类IMC来源的细胞系。未来,我们旨在研究xCT对犬类肿瘤中癌症干细胞特性维持的影响,并提出一种以xCT为靶点治疗犬类IMC的新治疗方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/218a/9281518/dcf5d3273255/jvetres-66-273-g001.jpg

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