Gong Y, Aimaiti A, He Z
Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Department of Transfusion Medicine, Southern Theater Command General Hospital, Guangzhou 510010, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2023 Dec 20;43(12):2006-2014. doi: 10.12122/j.issn.1673-4254.2023.12.03.
To investigate the effect of ARL67156, a small-molecule inhibitor of CD39, on cytotoxicity of natural killer (NK) cells against gastric cancer cells.
Human peripheral blood-derived primary NK cells isolated and purified using a magnetic bead antibody method were treated with 100 μmol/L ARL67156 for 24 h, and the signaling pathway of NK cell activation was detected by Western blotting. The level of interferon-γ (IFN-γ) in the supernatant of NK cells co-cultured with gastric cancer cells was detected using ELISA, and NK cell CD107a degranulation was measured with flow cytometry. The cytotoxicity of NK cells against co-cultured gastric cancer cells was evaluated using flow cytometry. In a nude mouse model bearing subcutaneous gastric cancer xenografts, the therapeutic effect of intravenous transfusion of NK cells and intraperitoneal injection of ARL67156 was assessed by measuring the changes in tumor volume.
(25.97 ± 5.69) % of peripheral blood NK cells from healthy individuals positive for CD39 expression. Treatment with ARL67156 significantly upregulated the activation molecules including NKG2D, DAP10, CD57, and CD16 and reduced the expressions of the inhibitory receptors TIGIT and KIR, thereby promoting the secretion of IFN-γ and CD107a degranulation in NK cells ( < 0.05). In both the and experiments, ARL67156 significantly enhanced the cytotoxicity of NK cells against gastric cancer cells ( < 0.05).
ARL67156 activates NK cells through the vav1-Syk signaling pathway to enhance their cytotoxicity against gastric cancer cells, which may serve as a new strategy for NK cell immunotherapy for gastric cancer.
研究CD39小分子抑制剂ARL67156对自然杀伤（NK）细胞对胃癌细胞细胞毒性的影响。
采用磁珠抗体法分离纯化人外周血来源的原代NK细胞，用100μmol/L ARL67156处理24小时，通过蛋白质免疫印迹法检测NK细胞活化的信号通路。采用酶联免疫吸附测定法检测与胃癌细胞共培养的NK细胞上清液中干扰素-γ（IFN-γ）水平，用流式细胞术检测NK细胞CD107a脱颗粒情况。采用流式细胞术评估NK细胞对共培养胃癌细胞的细胞毒性。在携带皮下胃癌异种移植瘤的裸鼠模型中，通过测量肿瘤体积变化评估静脉输注NK细胞和腹腔注射ARL67156的治疗效果。
健康个体外周血NK细胞中（25.97±5.69）% CD39表达呈阳性。用ARL67156处理显著上调了包括NKG2D、DAP10、CD57和CD16在内的活化分子，并降低了抑制性受体TIGIT和KIR的表达，从而促进NK细胞中IFN-γ的分泌和CD107a脱颗粒（<0.05）。在体内和体外实验中，ARL67156均显著增强了NK细胞对胃癌细胞的细胞毒性（<0.05）。
ARL67156通过vav1-Syk信号通路激活NK细胞，增强其对胃癌细胞的细胞毒性，这可能成为胃癌NK细胞免疫治疗的新策略。
