Visualizing the cancer stem-like properties of canine tumour cells with low proteasome activity
Visualizing the cancer stem-like properties of canine tumour cells with low proteasome activity
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蛋白酶体活性低的犬肿瘤细胞的癌症干细胞样特性可视化
DOI：
10.1111/vco.12779
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发表时间：
2022
影响因子：
2.1
通讯作者：
Masahiro Okumura 1
中科院分区：
文献类型：
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作者：
Koangyong Sung 1;Kenji Hosoya 2;Yusuke Murase 1;Tatsuya Deguchi 2;Sangho Kim 1;Takafumi Sunaga 1;Masahiro Okumura 1
癌症干细胞样细胞（Cancer stem - like cells, CSCs）导致多种肿瘤治疗失败；然而，由于难以识别和分离这一小群细胞，建立CSC靶向治疗一直受到阻碍。最近的研究表明，具有低蛋白酶体活性的肿瘤细胞显示CSC表型，可用于犬CSC成像。本研究可视化并揭示了犬细胞系HMPOS（骨肉瘤）和MegTCC（移行细胞癌）中具有低蛋白酶体活性的肿瘤细胞的CSC样特性。亲本细胞通过基因工程表达ZsGreen1，这是一种与犬鸟氨酸脱羧酶羧基末端脱羧酶连接的荧光蛋白，该酶以低蛋白酶体活性（ZsG+细胞）积累。对ZsG+细胞进行成像，并使用蛋白酶体抑制剂（MG‐132）确认该系统的作用模式，该抑制剂增加了ZsGreen1的荧光强度。根据细胞分裂、细胞周期、CSC标记物表达和致瘤性评价ZsG+细胞的CSC样特性。ZsG+细胞不对称分裂，G0/G1期细胞比例低；此外，ZsG+细胞表达CSC标记物如cd133，显示出较大的致瘤能力。组织病理学分析显示，ZsG+细胞广泛分布于来源于ZsG+细胞的肿瘤样本和肿瘤的增殖区域。本研究结果表明，具有低蛋白酶体活性的可视化犬肿瘤细胞具有CSC样表型，并且该可视化系统可用于鉴定和分离犬CSC。
Cancer stem‐like cells (CSCs) cause treatment failure in various tumours; however, establishing CSC‐targeted therapies has been hampered by difficulties in the identification and isolation of this small sub‐population of cells. Recent studies have revealed that tumour cells with low proteasome activity display a CSC phenotype that can be utilized to image CSCs in canines. This study visualizes and reveals the CSC‐like properties of tumour cells with low proteasome activity in HMPOS (osteosarcoma) and MegTCC (transitional cell carcinoma), which are canine cell lines. The parent cells were genetically engineered to express ZsGreen1, a fluorescent protein connected to the carboxyl‐terminal degron of canine ornithine decarboxylase that accumulates with low proteasome activity (ZsG+cells). ZsG+cells were imaged and the mode of action of this system was confirmed using a proteasome inhibitor (MG‐132), which increased the ZsGreen1 fluorescence intensity. The CSC‐like properties of ZsG+cells were evaluated on the basis of cell divisions, cell cycle, the expression of CSC markers and tumourigenicity. ZsG+cells underwent asymmetric divisions and had a low percentage of G0/G1 phase cells; moreover, ZsG+cells expressed CSC markers such asCD133and showed a large tumourigenic capability. In histopathological analysis, ZsG+cells were widely distributed in the tumour samples derived from ZsG+cells and in the proliferative regions of the tumours. The results of this study indicate that visualized canine tumour cells with low proteasome activity have a CSC‐like phenotype and that this visualization system can be utilized to identify and isolate canine CSCs.