美司钠靛抑制脑胶质瘤 U87 细胞的增殖，通过下调 PI3K/Akt 通路诱导细胞凋亡。

Meisoindigo inhibits cellular proliferation via down-regulation of the PI3K/Akt pathway and induces cellular apoptosis in glioblastoma U87 cells.

机构信息

1Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, China; 2Department of Neurosurgery, Wuhan Asia General Hospital, Wuhan, Hubei 430056, China.

Central laboratory, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, China.

出版信息

Acta Biochim Pol. 2021 May 23;68(2):309-315. doi: 10.18388/abp.2020_5581.

DOI:10.18388/abp.2020_5581

PMID:34022786

Abstract

OBJECTIVE

The current study was to explore whether meisoindigo was effective in suppressing proliferation and inducing apoptosis of human glioblastoma multiforme U87 cells and to explore its possible mechanisms.

METHOD

Morphological changes were observed by light microscopy. Cell counting kit-8 (CCK-8) assay was performed to detect cellular proliferation. Apoptosis was monitored by flow cytometry. Akt, phospho-Akt, PI3K, p65, phospho-p65 and apoptosis-related proteins caspase-3 and caspase-9 were examined by Western blotting assays. Immunofluorescence was used to evaluate level of P65 expression in cells.

RESULT

Meisoindigo inhibited the proliferation of U87 cells, and the inhibitory effect increased in a dose dependent manner. Moreover, meisoindigo exposure triggered an increase in the level of caspase-3 and caspase-9, supporting its role in the activation of apoptosis. Furthermore, meisoindigo reduced the expression of PI3K, Akt, phospho-Akt, NF-κB, p65 and phospho-p65 in U87 cells, and displacement of p65 from the nucleus to the cytoplasm.

CONCLUSION

Meisoindigo inhibits proliferation and induces apoptosis of U87 cells, probably through down-regulating the PI3K/Akt pathway and reducing nuclear translocation of NF-κB p65.

摘要

目的

本研究旨在探讨美司钠是否能有效抑制人多形性胶质母细胞瘤 U87 细胞的增殖并诱导其凋亡，并探讨其可能的机制。

方法

通过光镜观察形态学变化。用细胞计数试剂盒-8（CCK-8）检测细胞增殖。用流式细胞术监测细胞凋亡。用 Western blot 检测 Akt、磷酸化 Akt、PI3K、p65、磷酸化 p65 及凋亡相关蛋白 caspase-3 和 caspase-9。用免疫荧光法检测细胞中 P65 的表达水平。

结果

美司钠抑制 U87 细胞增殖，且抑制作用呈剂量依赖性。此外，美司钠诱导 caspase-3 和 caspase-9 水平升高，支持其在凋亡激活中的作用。此外，美司钠降低了 U87 细胞中 PI3K、Akt、磷酸化 Akt、NF-κB、p65 和磷酸化 p65 的表达，并使 p65 从核内易位到细胞质。