Faculty of Science, Department of Biology, Istanbul University, Istanbul, Turkey.
Protoplasma. 2024 Mar;261(2):281-291. doi: 10.1007/s00709-023-01896-w. Epub 2023 Oct 6.
Insulinoma is an excessive insulin-released beta cell tumor. Pancreas cancer is one of the deadliest malignant neoplasms. Exosomes are secreted cell membrane vesicles containing a large number of proteins, lipids, and nucleic acids. The aim of this study is to investigate the effects of exosomes on two cell lines of benign and malignant character. For the first time, exosomes were isolated from pancreatic island-derived progenitor cells (PID-PCs) and applied to INS-1 and MiaPaCa-2 cells. In addition, exosomes isolated from PID-PC, MiaPaca-2, and INS-1 cells were characterized in order to compare their sizes with other previously isolated exosomes. Alix, TSG101, CD9, and CD81 were analyzed. The size and concentration of exosomes and the cell viability were detected. The cells were marked with HSP90, HSF-1, Kaspaz-8, Active-Kaspaz-3, Beclin, and p-Bcl-2. The cell cytotoxicity and insulin levels kit were measured. Alix in all exosomes, and PID-PC, MiaPaca-2 cell lysates; TSG101 in PID-PC and MiaPaca-2 cell lysates; CD9 in INS-1 exosomes were detected. The dimensions of isolated exosomes were 103.6 ± 28.6 nm, 100.7 ± 10 nm, and 147.2 ± 12.3 nm for PID-PCs, MiaPaca-2, and INS-1 cells. The cell viability decreased and HSP90 increased in the MiaPaca-2 cells. The HSF-1 was higher in the control MiaPaca-2 cell compared to the control INS-1 cell, and the exosome-treated MiaPaca-2 cell compared to the exosome-treated INS-1 cell. Beclin and p-Bcl-2 were decreased in the exosome-treated MiaPaca-2 cells. The insulin level in the cell lysates increased compared to cell secretion in INS-1 cells. In conclusion, exosomes isolated from the PID-PC caused cell death in the MiaPaca-2 cells in a time- and dose-dependent manner. The IC50 value determined for MiaPaca-2 cells has no effect on cell viability in INS-1 cells, which best mimics pancreatic beta cells and can be used instead of healthy pancreatic beta cells. Isolated exosomes can kill cancer cells without damaging healthy cells.
胰岛细胞瘤是一种过度分泌胰岛素的β细胞肿瘤。胰腺癌是最致命的恶性肿瘤之一。外泌体是一种含有大量蛋白质、脂质和核酸的细胞分泌的膜囊泡。本研究的目的是研究外泌体对两种良性和恶性特征的细胞系的影响。本文首次从胰岛源性祖细胞（PID-PC）中分离出外泌体，并应用于 INS-1 和 MiaPaCa-2 细胞。此外，还对从 PID-PC、MiaPaca-2 和 INS-1 细胞中分离出的外泌体进行了表征，以比较其与其他先前分离的外泌体的大小。分析了 Alix、TSG101、CD9 和 CD81。检测外泌体的大小和浓度以及细胞活力。用 HSP90、HSF-1、Kaspaz-8、活性 Kaspaz-3、Beclin 和 p-Bcl-2 标记细胞。测量细胞细胞毒性和胰岛素水平试剂盒。在所有外泌体和 PID-PC、MiaPaca-2 细胞裂解物中检测到 Alix；在 PID-PC 和 MiaPaca-2 细胞裂解物中检测到 TSG101；在 INS-1 外泌体中检测到 CD9。从 PID-PC、MiaPaca-2 和 INS-1 细胞中分离出的外泌体的尺寸分别为 103.6 ± 28.6nm、100.7 ± 10nm 和 147.2 ± 12.3nm。MiaPaca-2 细胞的细胞活力下降，HSP90 增加。与对照 INS-1 细胞相比，对照 MiaPaca-2 细胞中的 HSF-1 更高，与对照 INS-1 细胞相比，外泌体处理的 MiaPaca-2 细胞中的 HSF-1 更高。外泌体处理的 MiaPaca-2 细胞中的 Beclin 和 p-Bcl-2 减少。与 INS-1 细胞的细胞分泌相比，细胞裂解物中的胰岛素水平增加。总之，PID-PC 分离的外泌体以时间和剂量依赖的方式导致 MiaPaca-2 细胞死亡。确定的 MiaPaca-2 细胞的 IC50 值对 INS-1 细胞的细胞活力没有影响，INS-1 细胞最能模拟胰腺β细胞，可替代健康的胰腺β细胞。分离的外泌体可以杀死癌细胞而不损伤健康细胞。
