Identification of rhodamine 123-positive stem cell subpopulations in canine hepatocellular carcinoma cells

Itoh,Harumichi; Nishikawa,Shimpei; Haraguchi,Tomoya; Arikawa,Yu; Hiyama,Masato; Iseri,Toshie; Itoh,Yoshiki; Nakaichi,Munekazu; Taura,Yasuho; Tani,Kenji; Itamoto,Kazuhito

doi:10.3892/br.2017.925

Identification of rhodamine 123-positive stem cell subpopulations in canine hepatocellular carcinoma cells

Authors:
- Harumichi Itoh
- Shimpei Nishikawa
- Tomoya Haraguchi
- Yu Arikawa
- Masato Hiyama
- Toshie Iseri
- Yoshiki Itoh
- Munekazu Nakaichi
- Yasuho Taura
- Kenji Tani
- Kazuhito Itamoto
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Affiliations: Department of Small Animal Clinical Science, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi City, Yamaguchi 753‑8511, Japan, Department of Veterinary Surgery, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi City, Yamaguchi 753‑8511, Japan, Department of Veterinary Radiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi City, Yamaguchi 753‑8511, Japan
Published online on: June 7, 2017 https://doi.org/10.3892/br.2017.925
Pages: 73-78

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Abstract

The majority of cases of chemotherapy for hepatocellular carcinoma (HCC) are not effective in human or veterinary medicine due to resistance against anticancer agents. In human medicine, hepatocellular carcinoma stem cells (HCSCs) were recently identified as cytokeratin 19 (CK19)‑, cluster of differentiation (CD)‑44‑, and CD133‑positive. However, there are few previous reports regarding canine HCSC (cHCSC). Additionally, to the best of our knowledge, the chemoresistance against anticancer agents of these cHCSCs has not been investigated. In the present study staining of cHCSCs was performed with rhodamine 123, a low‑toxicity fluorescent dye for mitochondria, by flow cytometry. There were two subpopulations in the HCC cell line defined by their higher (RhoHi) and lower (RhoLo) fluorescence intensity of rhodamine 123. The RhoHi subpopulation demonstrated a higher Nanog gene expression, sphere‑forming ability, and chemoresistance against gemcitabine. However, there was no significant difference between RhoHi and RhoLo regarding the proliferation rate and chemoresistance against mitoxantrone and doxorubicin. The present results indicate that the expression of rhodamine 123 identifies different stem cell subpopulations in a canine HCC cell line.

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