Abbasi Sana, Movassaghpour AliAkbar, Soleimani Masoud, Asghari Molabashi Zahra
Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Int J Hematol Oncol Stem Cell Res. 2025 Apr 1;19(2):126-137. doi: 10.18502/ijhoscr.v19i2.18550.
: Most cancers are treated through chemotherapy and radiotherapy. However, these methods have limitations due to cancer cells evading immune detection, prompting researchers to explore alternatives such as immunotherapy. Nonetheless, cancer cells can weaken the immune response, necessitating improvements in immunotherapy methods. Exosomes, tiny cell-derived nanoparticles, reflect the traits of their originating cells. Natural Killer NK cells produce exosomes comprising perforin, granzyme, Fas-L, etc. The small size, proximity to tumors, and stability of these exosomes enable easy absorption by cancer cells. This study demonstrates that IL-15 impacts NK-derived exosomes, enhancing their ability to kill cancer cells. With the addition of 100 nanograms per milliliter of IL-15 to NK-92 cell culture, the cells are incubated for 48 hours. Exosomes are then isolated from treated and non-treated NK-92 cell lines through the ultracentrifuge method. After isolation, different concentrations of exosomes from both groups are added to HL-60 cells for treatment. After 24 hours, the apoptosis rate is assessed through the Annexin-V method. Increased light absorption in the BCA test, along with thicker bands of CD63 and CD81 in the Western blotting test, indicates a higher yield of exosomes after adding IL-15 to the source cells. The low p-value from the t-test demonstrates that exosomes derived from stimulated NK cells are more cytotoxic than those from the control group. Further, two-way ANOVA confirms differences between the control and treatment groups at each concentration, and Welch's t-test proves that all differences in the ANOVA test are significant. This article presents evidence that exosomes obtained from IL-15-induced NK cells not only increase in quantity but also demonstrate significant cytotoxicity against leukemic cells compared to exosomes obtained from non-stimulated NK cells.
大多数癌症通过化疗和放疗进行治疗。然而，由于癌细胞能够逃避免疫检测，这些方法存在局限性，这促使研究人员探索诸如免疫疗法等替代方法。尽管如此，癌细胞会削弱免疫反应，因此需要改进免疫疗法。外泌体是源自细胞的微小纳米颗粒，反映了其来源细胞的特征。自然杀伤（NK）细胞产生包含穿孔素、颗粒酶、Fas-L等的外泌体。这些外泌体体积小、靠近肿瘤且稳定性好，能够被癌细胞轻易吸收。本研究表明，白细胞介素-15（IL-15）会影响NK细胞衍生的外泌体，增强其杀死癌细胞的能力。在NK-92细胞培养中添加每毫升100纳克的IL-15，将细胞孵育48小时。然后通过超速离心法从处理过和未处理过的NK-92细胞系中分离出外泌体。分离后，将两组不同浓度的外泌体添加到HL-60细胞中进行处理。24小时后，通过膜联蛋白-V法评估凋亡率。BCA测试中光吸收增加，以及蛋白质免疫印迹测试中CD63和CD81条带变厚，表明向来源细胞中添加IL-15后外泌体产量更高。t检验的低p值表明，刺激后的NK细胞衍生的外泌体比对照组的外泌体具有更强的细胞毒性。此外，双向方差分析证实了对照组和处理组在每个浓度下的差异，而韦尔奇t检验证明方差分析测试中的所有差异均具有显著性。本文提供的证据表明，与未刺激的NK细胞获得的外泌体相比，从IL-15诱导的NK细胞获得的外泌体不仅数量增加，而且对白血病细胞表现出显著的细胞毒性。
