Lv Wen-wen, Qin Si-ning, Chen Cong-qin, Zhang Jin-jie, Ren Tian-shu, Xu Yong-nan, Zhao Qing-chun
1] Department of Pharmacy, General Hospital of Shenyang Military Area Command, Shenyang 110840, China [2] Shenyang Pharmaceutical University, Shenyang 110016, China.
Shenyang Pharmaceutical University, Shenyang 110016, China.
Acta Pharmacol Sin. 2014 May;35(5):625-35. doi: 10.1038/aps.2013.194.
2-(4,6-Dimethoxy-1,3-dioxoisoindolin-2-yl) ethyl 2-chloroacetate (QSN-10c) is one of isoindolone derivatives with antiproliferative activity against human umbilical vein endothelial cells (HUVECs). The aim of this study was to investigate its antitumor activity in vitro and anti-angiogenic effects in vitro and in vivo.
K562 leukemic cells and HUVECs were used for in vitro studies. Cell viability was examined using MTT assay. Cell apoptosis and mitochondrial transmembrane potential (Δψm) were detected with flow cytometry. Tube formation and migration of HUVECs were studied using two-dimensional Matrigel assay and wound-healing migration assay, respectively. VEGF levels were analyzed with RT-PCR and Western blotting. A zebrafish embryo model was used for in vivo anti-angiogenic studies. The molecular mechanisms for apoptosis in K562 cells and antiangiogenesis were measured with Western blotting.
In antitumor activity studies, QSN-10c suppressed the viability of K562 cells and induced apoptosis in dose- and time-dependent manners. Furthermore, QSN-10c dose-dependently decreased the Δψm in K562 cells, increased the release of cytochrome c and the level of Bax, and decreased the level of Bcl-2, suggesting that QSN-10c-induced apoptosis of K562 cells was mediated via the mitochondrial apoptotic pathway. In anti-angiogenic activity studies, QSN-10c suppressed the viability of HUVECs and induced apoptosis in dose dependent manners. QSN-10c treatment did not alter the Δψm in HUVECs, but dose-dependently inhibited the expression of VEGF, inhibited the tube formation and cell migration in vitro, and significantly suppressed the number of ISVs in zebrafish embryos in vivo. Furthermore, QSN-10c dose-dependently suppressed the phosphorylation of AKT and GSK3β in both HUVECs and K562 cells.
QSN-10c is a novel antitumor compound that exerts both antitumor and anti-angiogenic effects via inhibiting the PI3K/AKT/GSK3β signaling pathway.
2-(4,6-二甲氧基-1,3-二氧代异吲哚啉-2-基)乙基 2-氯乙酸酯（QSN-10c）是一种对人脐静脉内皮细胞（HUVECs）具有抗增殖活性的异吲哚酮衍生物。本研究旨在探讨其体外抗肿瘤活性以及体外和体内的抗血管生成作用。
使用 K562 白血病细胞和 HUVECs 进行体外研究。采用 MTT 法检测细胞活力。通过流式细胞术检测细胞凋亡和线粒体跨膜电位（Δψm）。分别使用二维基质胶试验和伤口愈合迁移试验研究 HUVECs 的管腔形成和迁移。用 RT-PCR 和蛋白质印迹法分析 VEGF 水平。使用斑马鱼胚胎模型进行体内抗血管生成研究。用蛋白质印迹法检测 K562 细胞凋亡和抗血管生成的分子机制。
在抗肿瘤活性研究中，QSN-10c 以剂量和时间依赖性方式抑制 K562 细胞的活力并诱导凋亡。此外，QSN-10c 剂量依赖性地降低 K562 细胞中的 Δψm，增加细胞色素 c 的释放和 Bax 水平，并降低 Bcl-2 水平，表明 QSN-10c 诱导的 K562 细胞凋亡是通过线粒体凋亡途径介导的。在抗血管生成活性研究中，QSN-10c 以剂量依赖性方式抑制 HUVECs 的活力并诱导凋亡。QSN-10c 处理未改变 HUVECs 中的 Δψm，但剂量依赖性地抑制 VEGF 的表达，抑制体外管腔形成和细胞迁移，并显著抑制体内斑马鱼胚胎中节间血管（ISV）的数量。此外，QSN-10c 剂量依赖性地抑制 HUVECs 和 K562 细胞中 AKT 和 GSK3β 的磷酸化。
QSN-10c 是一种新型抗肿瘤化合物，通过抑制 PI3K/AKT/GSK3β 信号通路发挥抗肿瘤和抗血管生成作用。
