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MAP30 promotes apoptosis of U251 and U87 cells by suppressing the LGR5 and Wnt/β-catenin signaling pathway and enhancing Smac expression

机译:MAP30通过抑制LGR5和Wnt /β-catenin信号通路并增强Smac表达来促进U251和U87细胞凋亡。

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摘要

Significant antitumor activity of Momordica anti-human immunodeficiency virus protein of 30 kDa (MAP30) purified from Momordica charantia 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.
机译:从苦瓜中提纯的苦瓜抗人免疫缺陷病毒蛋白30 kDa(MAP30)的显着抗肿瘤活性已成为先前研究的主题。然而,MAP30对恶性神经胶质瘤细胞的有效机制尚未阐明。本研究的目的是研究MAP30对U87和U251细胞系的作用和机制。使用Cell Counting Kit-8测定法,伤口愈合测定法和Transwell测定法检测在不同时间段内对不同浓度的MAP30(0.5、1、2、4、8和16 µM)处理的U87和U251细胞的影响。 MAP30以剂量和时间依赖性方式显着抑制每种细胞系的增殖,迁移和侵袭。流式细胞仪和荧光染色表明,在MAP30处理后的两个研究细胞系中,细胞凋亡增加,细胞周期停滞在S期。 Western印迹分析表明Wnt /β-catenin信号通路中富含亮氨酸重复的G蛋白偶联受体5(LGR5)表达和关键蛋白明显减少,而第二个线粒体衍生的半胱天冬酶(Smac)激活剂MAP30处理以相同的方式使蛋白质表达显着增加。这些结果表明,MAP30通过抑制LGR5和Wnt /β-catenin信号通路,并以剂量​​和时间依赖性方式增强Smac表达,从而明显诱导U87和U251细胞株的凋亡。

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