Department of Oncology, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong, 510282, China; Department of Medical Oncology, Second Clinical Medicine College of Jinan University, Shenzhen People's Hospital, Shenzhen 518020, China.
Department of Medical Oncology, Second Clinical Medicine College of Jinan University, Shenzhen People's Hospital, Shenzhen 518020, China.
Pathol Res Pract. 2019 Jun;215(6):152379. doi: 10.1016/j.prp.2019.03.008. Epub 2019 Mar 11.
Epidemiological studies have illustrated that regular aspirin consumption may decrease the risk of non-small cell lung cancer (NSCLC). The present study aims to investigate the mechanism of aspirin-induced inhibition of NSCLC development during hypoxia.
A549 cells were pre-treated with the vehicle control or aspirin and then subjected to hypoxic culture. Cell viability was monitored by CCK-8 assay, and flow cytometry was performed to detect cell cycle distributions, apoptosis, and proportion of cancer stem cells (CSCs). Flow cytometric cell sorting was used to separate CSCs. Quantitative reverse transcription-polymerase chain reaction and Western blot were used to detect the mRNA and protein levels of stem cell markers and the related signaling molecules. The abundance of prostaglandin E2 was detected by enzyme-linked immunosorbent assay. Exosomes in the cell culture medium were isolated using ExoQuick, and the number of exosomes was quantified by the EXOCET exosome quantification assay kit. Cell migration and angiogenesis were monitored by transwell migration assay and in vitro angiogenesis experiments.
Aspirin inhibited cell proliferation and induced G2/M cell cycle arrest in hypoxic A549 cells; it also inhibited hypoxia-enhanced stemness in both A549 and ALDH cells. The drug reduced hypoxia-enhanced numbers of exosomes in A549 cells and exerted negative effects on the hypoxia-mediated up-regulation of exosomal HIF-1α/COX-2 and expression of exosomal miR-135b and miR-210. While hypoxic-induced exosomes can promote the proliferation, migration, and angiogenesis of other A549 cells, aspirin can weaken this promotion by reducing the amount of exosome secreted and changing exosome contents.
Aspirin inhibits the hypoxia-induced stemness, hypoxic-mediated exosome release, and malignant paracrine effects of A549 cells.
流行病学研究表明，规律服用阿司匹林可能降低非小细胞肺癌（NSCLC）的发病风险。本研究旨在探讨阿司匹林在低氧环境下抑制 NSCLC 发展的作用机制。
用载剂对照或阿司匹林预处理 A549 细胞，然后进行低氧培养。用 CCK-8 法检测细胞活力，用流式细胞术检测细胞周期分布、凋亡和肿瘤干细胞（CSC）比例。用流式细胞术分选 CSCs。用定量逆转录聚合酶链反应和 Western blot 检测干细胞标志物和相关信号分子的 mRNA 和蛋白水平。用酶联免疫吸附试验检测前列腺素 E2 的丰度。用 ExoQuick 分离细胞培养上清中的外泌体，用 EXOCET 外泌体定量检测试剂盒定量外泌体数量。用 Transwell 迁移实验和体外血管生成实验监测细胞迁移和血管生成。
阿司匹林抑制低氧 A549 细胞的增殖，诱导 G2/M 细胞周期阻滞；还抑制 A549 和 ALDH 细胞中低氧增强的干性。该药物减少低氧增强的 A549 细胞中外泌体数量，并对低氧介导的外泌体 HIF-1α/COX-2 上调和外泌体 miR-135b 和 miR-210 表达产生负向影响。虽然低氧诱导的外泌体可以促进其他 A549 细胞的增殖、迁移和血管生成，但阿司匹林可以通过减少外泌体分泌量和改变外泌体内容物来减弱这种促进作用。
阿司匹林抑制低氧诱导的干性、低氧介导的外泌体释放和 A549 细胞的恶性旁分泌效应。
