Viral Disease Research Division, Animal and Plant Quarantine Agency, 177 Hyeoksin 8-ro, Gimcheon, Gyeongsangbuk-do 39660, Republic of Korea.
Division of Regenerative Medicine Safety Control, Department of Chronic Disease Convergence Research, Korea National Institute of Health, Korea Disease Control and Prevention Agency, 202 Osongsaengmyeong 2-ro, Cheongju, Chungcheongbuk-do 28159, Republic of Korea.
Biomed Res Int. 2021 Sep 4;2021:6690704. doi: 10.1155/2021/6690704. eCollection 2021.
Natural killer (NK) cells are key immune cells engaged in fighting infection and malignant transformation. In this study, we found that canine NK cell-derived exosomes (NK-exosomes) separated from activated cytotoxic NK cell supernatants express specific markers including CD63, CD81, Alix, HSP70, TSG101, Perforin 1, and Granzyme B. We examined the antitumor effects of NK-exosomes in an experimental murine mammary tumor model using REM134 canine mammary carcinoma cell line. We observed changes in tumor size, tumor initiation, progression, and recurrence-related markers in the control, tumor group, and NK-exosome-treated tumor group. We found that the tumor size in the NK-exosome-treated tumor group decreased compared with that of the tumor group in the REM134-driven tumorigenic mouse model. We observed significant changes including the expression of tumorigenesis-related markers, such as B cell-specific Moloney murine leukemia virus insertion site 1 (Bmi-1), vascular endothelial growth factor (VEGF), matrix metallopeptidase-3 (MMP-3), interleukin-1 (IL-1), IL-6, tumor necrosis factor- (TNF-), multidrug resistance protein (MDR), tumor suppressor protein p53 (p53), proliferating cell nuclear antigen (PCNA), and the apoptotic markers, B cell lymphoma-2 associated X (Bax) and B cell lymphoma-extra large (Bcl-xL) belonging to the Bcl-2 family, in the tumor group compared with those in the control group. The expression of CD133, a potent cancer stem cell marker, was significantly higher than that of the control. By contrast, the NK-exosome-treated tumor group exhibited a significant reduction in Bmi-1, MMP-3, IL-1, IL-6, TNF-, Bax, Bcl-xL, and PCNA expression compared with that in the tumor group. Furthermore, the expression of CD133, which mediates tumorigenesis, was significantly decreased in the NK-exosome-treated tumor group compared with that in the tumor group. These findings indicate that canine NK-exosomes represent a promising therapeutic tool against canine solid tumors, including mammary carcinoma.
自然杀伤 (NK) 细胞是参与抗感染和恶性转化的关键免疫细胞。在这项研究中，我们发现从激活的细胞毒性 NK 细胞上清液中分离的犬 NK 细胞衍生的外泌体 (NK-exosomes) 表达特定的标记物，包括 CD63、CD81、Alix、HSP70、TSG101、Perforin 1 和 Granzyme B。我们使用 REM134 犬乳腺肿瘤细胞系检查了 NK-exosomes 在实验性乳腺肿瘤模型中的抗肿瘤作用。我们观察了对照组、肿瘤组和 NK-exosome 处理的肿瘤组中肿瘤大小、肿瘤起始、进展和与复发相关的标记物的变化。我们发现，与 REM134 驱动的致瘤小鼠模型中的肿瘤组相比，NK-exosome 处理的肿瘤组的肿瘤大小减小。我们观察到了显著的变化，包括肿瘤发生相关标记物的表达，如 B 细胞特异性 Moloney 鼠白血病病毒插入位点 1 (Bmi-1)、血管内皮生长因子 (VEGF)、基质金属蛋白酶-3 (MMP-3)、白细胞介素-1 (IL-1)、白细胞介素-6 (IL-6)、肿瘤坏死因子- (TNF-)、多药耐药蛋白 (MDR)、肿瘤抑制蛋白 p53 (p53)、增殖细胞核抗原 (PCNA) 和属于 Bcl-2 家族的凋亡标记物 B 细胞淋巴瘤-2 相关 X (Bax) 和 B 细胞淋巴瘤-额外大 (Bcl-xL)。肿瘤组中与对照组相比，CD133 的表达显著更高，CD133 是一种有效的癌症干细胞标记物。相比之下，NK-exosome 处理的肿瘤组中 Bmi-1、MMP-3、IL-1、IL-6、TNF-、Bax、Bcl-xL 和 PCNA 的表达与肿瘤组相比均显著降低。此外，NK-exosome 处理的肿瘤组中与肿瘤发生有关的 CD133 的表达也明显低于肿瘤组。这些发现表明，犬 NK-exosomes 是一种有前途的治疗犬实体肿瘤，包括乳腺癌的治疗工具。
