沉默肿瘤内在的 CD73 可增强 NSCLC 的化疗敏感性，并增强顺铂的抗肿瘤作用：一项体外研究。
Silencing tumor-intrinsic CD73 enhances the chemosensitivity of NSCLC and potentiates the anti-tumoral effects of cisplatin: An in vitro study.
机构信息
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
出版信息
Biomed Pharmacother. 2022 Jan;145:112370. doi: 10.1016/j.biopha.2021.112370. Epub 2021 Nov 30.
AIMS
Besides suppressing anti-tumoral immune responses, tumor-intrinsic inhibitory immune checkpoints have been implicated in tumor development. Herein, we aimed to investigate the significance of tumor-intrinsic CD73, as an inhibitory immune checkpoint, in non-small cell lung cancer (NSCLC) development and propose a novel therapeutic approach.
MAIN METHODS
We investigated the cell viability, chemosensitivity, apoptosis, migration, and the cell cycle of A-549 and NCI-H1299 following treatment with cisplatin and CD73-small interfering RNA (siRNA) transfection. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to study the viability of studied groups and chemosensitivity of tumoral cells. Flow cytometry and 4',6-diamidino-2-phenylindole (DAPI) staining were used to investigate the apoptosis of NSCLC cells. Flow cytometry and the wound-healing assay were used to investigate the cell cycle and migration of NSCLC cells, respectively. The mRNA expression levels of c-Myc, caspase 3, ROCK, and MMP-9 were investigated to study the underlying molecular mechanism.
KEY FINDINGS
CD73-siRNA transfection has significantly decreased the cell viability and enhanced the chemosensitivity of A-549 and NCI-H1299 cells to cisplatin. CD73-siRNA has considerably stimulated apoptosis, arrested the cell cycle, inhibited tumor migration, downregulated the mRNA expression of c-Myc, MMP-9, and ROCK, and upregulated caspase 3 expression in NSCLC cells. Besides, combined cisplatin therapy with CD73-siRNA transfection has potentiated the aforementioned anti-tumoral effects of cisplatin on NSCLC cells.
SIGNIFICANCE
Besides suppressing anti-tumoral immune responses, tumor-intrinsic CD73 can facilitate NSCLC development, and the combined cisplatin therapy with CD73-siRNA transfection can substantially enhance the chemosensitivity of NSCLC to cisplatin and potentiates cisplatin-induced anti-tumoral effects on NSCLC.
目的
除了抑制抗肿瘤免疫反应外，肿瘤内在的抑制性免疫检查点也被认为与肿瘤的发展有关。在此，我们旨在研究肿瘤内在的 CD73（一种抑制性免疫检查点）在非小细胞肺癌（NSCLC）发展中的意义，并提出一种新的治疗方法。
方法
我们用顺铂处理 A-549 和 NCI-H1299 细胞，并进行 CD73-小干扰 RNA（siRNA）转染，检测细胞活力、化疗敏感性、细胞凋亡、迁移和细胞周期的变化。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐（MTT）法检测研究组细胞活力和肿瘤细胞化疗敏感性。用流式细胞术和 4'，6-二脒基-2-苯基吲哚（DAPI）染色检测 NSCLC 细胞凋亡。用流式细胞术和划痕实验分别检测 NSCLC 细胞的细胞周期和迁移。用实时定量 PCR 检测 c-Myc、半胱天冬酶 3、ROCK 和 MMP-9 的 mRNA 表达水平，研究潜在的分子机制。
主要发现
CD73-siRNA 转染显著降低了 A-549 和 NCI-H1299 细胞的活力，并增强了它们对顺铂的化疗敏感性。CD73-siRNA 可显著促进 NSCLC 细胞凋亡，阻滞细胞周期，抑制肿瘤迁移，下调 c-Myc、MMP-9 和 ROCK 的 mRNA 表达，上调半胱天冬酶 3 的表达。此外，顺铂联合 CD73-siRNA 转染增强了顺铂对 NSCLC 细胞的上述抗肿瘤作用。
意义
除了抑制抗肿瘤免疫反应外，肿瘤内在的 CD73 还可以促进 NSCLC 的发展，顺铂联合 CD73-siRNA 转染可以显著提高 NSCLC 对顺铂的化疗敏感性，并增强顺铂对 NSCLC 的抗肿瘤作用。
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