Tugutova Elena A, Tamkovich Svetlana N, Patysheva Marina R, Afanas’ev Sergey G, Tsydenova Anastasia A, Grigor’eva Alina E, Kolegova Elena S, Kondakova Irina V, Yunusova Natalia V
Cancer Research Institute, Тomsk National Research Medical Center, Russian Academy of Science, Tomsk, Russia. Email:
Institute of Chemical Biology and Fundamental Medicine, SB RAS, Novosibirsk, Russia.
Asian Pac J Cancer Prev. 2019 Mar 26;20(3):809-815. doi: 10.31557/APJCP.2019.20.3.809.
Purpose: Exosomal proteases are important in regulation of molecular signaling from growth factor receptors and adhesion molecules and also the regulation of cell motility and protein folding. The aim of this study was to evaluate the level of ADAM10, ADAM17 and 20S proteasomes in exosomes isolated from colorectal cancer patients (CRCPs) in relation with clinical and histopathological parameters. Methods: Blood plasma exosomes of 60 CRCPs at stage T2-4N0-2M0-1 and 10 control subjects (CSs) with colorectal polyps were isolated using ultrafiltration in combination with ultracentrifugation. The level of tetraspanin-associated (ADAM20 and ADAM17) and tetraspanin-non-associated (20S proteasome) proteases were evaluated by flow cytometry and western blot analysis. Results: The ADAM10-/ ADAM17- population predominated in plasma exosomes of CRCPs and the level of ADAM10+ exosomes was significantly higher in exosomes of CSs compared with CRCPs. No difference was found between subpopulations of ADAM10/ADAM17 exosomes and level of exosomal 20S proteasomes in terms of sex, age and tumor grade. Simultaneous decrease of ADAM10+/ADAM17-subpopulation of exosomes and level of exosomal 20S proteasomes in patients with metastatic CRC was observed compared with patients with non-metastatic CRC. The level of ADAM17+ exosomes significantly reduced in exosomes of CRCPs with metabolic syndrome compared to CRCPs without metabolic syndrome( 3.97±0.71 (%) vs. 13.04±1.34 (%), respectively (p<0.05). A decrease in the 20S proteasomes level in plasma exosomes was revealed in CRCPs with metabolic syndrome compared with CRCPs without metabolic disorders ( 1.90±0.25 (r.u.) vs. 2.92±0.42 (r.u.) respectively( (p<0.05). Conclusion: According to findings of this study, it seems that exosomal proteases can be promising molecular predictors of hematogenous metastasis in patients with non-metastatic CRC. Further studies on subpopulation composition of exosomes CRCPs are need for elucidating the role of tetraspanin-associated and tetraspanin-non-associated exosomal proteases in CRC development and progression.
外泌体蛋白酶在调节生长因子受体和黏附分子的分子信号传导以及细胞运动和蛋白质折叠的调节中起着重要作用。本研究的目的是评估从结直肠癌患者（CRCPs）中分离出的外泌体中ADAM10、ADAM17和20S蛋白酶体的水平，并将其与临床和组织病理学参数相关联。方法：采用超滤结合超速离心法，从60例处于T2-4N0-2M0-1期的CRCPs患者和10例患有大肠息肉的对照受试者（CSs）的血浆中分离出外泌体。通过流式细胞术和蛋白质印迹分析评估四跨膜蛋白相关蛋白酶（ADAM20和ADAM17）和四跨膜蛋白非相关蛋白酶（20S蛋白酶体）的水平。结果：CRCPs患者血浆外泌体中以ADAM10-/ADAM17-群体为主，与CRCPs相比，CSs外泌体中ADAM10+外泌体的水平显著更高。在ADAM10/ADAM17外泌体亚群和外泌体20S蛋白酶体水平方面，在性别、年龄和肿瘤分级上未发现差异。与非转移性结直肠癌患者相比，转移性结直肠癌患者外泌体中ADAM10+/ADAM17-亚群和外泌体20S蛋白酶体水平同时降低。与无代谢综合征的CRCPs相比，患有代谢综合征的CRCPs外泌体中ADAM17+外泌体水平显著降低（分别为3.97±0.71（%）和13.04±1.34（%），p<0.05）。与无代谢紊乱的CRCPs相比，患有代谢综合征的CRCPs血浆外泌体中20S蛋白酶体水平降低（分别为1.90±0.25（相对单位）和2.92±0.42（相对单位），p<0.05）。结论：根据本研究结果，似乎外泌体蛋白酶可能是非转移性结直肠癌患者血行转移的有前景的分子预测指标。需要进一步研究CRCPs外泌体的亚群组成，以阐明四跨膜蛋白相关和四跨膜蛋白非相关外泌体蛋白酶在结直肠癌发生和发展中的作用。
