Jiang Yilin, Miao Junjie, Wang Dongliang, Zhou Jingru, Liu Bo, Jiao Feng, Liang Jiangfeng, Wang Yangshuo, Fan Cungang, Zhang Qingjun
Department of Neurosurgery, Peking University People's Hospital, Peking University, Beijing 100044, P.R. China.
Department of Neurosurgery, Peking University International Hospital, Beijing 102206, P.R. China.
Oncol Lett. 2018 Apr;15(4):5833-5840. doi: 10.3892/ol.2018.8073. Epub 2018 Feb 16.
Significant antitumor activity of anti-human immunodeficiency virus protein of 30 kDa (MAP30) purified from has been the subject of previous research. However, the effective mechanism of MAP30 on malignant glioma cells has not yet been clarified. The aim of the present study was to investigate the effects and mechanism of MAP30 on U87 and U251 cell lines. A Cell Counting Kit-8 assay, wound healing assay and Transwell assay were used to detect the effects on U87 and U251 cells treated with different concentrations of MAP30 (0.5, 1, 2, 4, 8 and 16 µM) over different periods of time. Proliferation, migration and invasion of each cell line were markedly inhibited by MAP30 in a dose- and time-dependent manner. Flow cytometry and fluorescence staining demonstrated that apoptosis increased and the cell cycle was arrested in S-phase in the two investigated cell lines following MAP30 treatment. Western blot analysis demonstrated that leucine-rich-repeat-containing G-protein-coupled receptor 5 (LGR5) expression and key proteins in the Wnt/β-catenin signaling pathway were apparently decreased, whereas second mitochondria-derived activator of caspase (Smac) protein expression significantly increased with MAP30 treatment in the same manner. These results suggest that MAP30 markedly induces apoptosis in U87 and U251 cell lines by suppressing LGR5 and the Wnt/β-catenin signaling pathway, and enhancing Smac expression in a dose- and time-dependent manner.
从[具体来源]纯化的30 kDa抗人免疫缺陷病毒蛋白(MAP30)具有显著的抗肿瘤活性,这是先前研究的主题。然而,MAP30对恶性胶质瘤细胞的有效作用机制尚未阐明。本研究的目的是探讨MAP30对U87和U251细胞系的影响及其机制。采用细胞计数试剂盒-8法、伤口愈合试验和Transwell试验,检测不同浓度(0.5、1、2、4、8和16 μM)的MAP30在不同时间段处理U87和U251细胞后的效果。MAP30以剂量和时间依赖性方式显著抑制各细胞系的增殖、迁移和侵袭。流式细胞术和荧光染色表明,MAP30处理后,所研究的两种细胞系的凋亡增加,细胞周期停滞在S期。蛋白质印迹分析表明,富含亮氨酸重复序列的G蛋白偶联受体5(LGR5)的表达以及Wnt/β-连环蛋白信号通路中的关键蛋白明显降低,而第二线粒体衍生的半胱天冬酶激活剂(Smac)蛋白表达则以相同方式随MAP30处理显著增加。这些结果表明,MAP30通过抑制LGR5和Wnt/β-连环蛋白信号通路,并以剂量和时间依赖性方式增强Smac表达,从而显著诱导U87和U251细胞系凋亡。