盐酸小檗胺通过 caspase 3 依赖性诱导胶质细胞瘤生长停滞。

pDok2, caspase 3 dependent glioma cell growth arrest by nitidine chloride.

机构信息

Department of Biotechnology and Bioinformatics, School of Life Sciences, University of Hyderabad, Hyderabad, Telangana State, India.

出版信息

Pharmacol Rep. 2018 Feb;70(1):48-54. doi: 10.1016/j.pharep.2017.07.013. Epub 2017 Jul 29.

DOI:10.1016/j.pharep.2017.07.013

PMID:29329030

Abstract

BACKGROUND

Nitidine chloride (NC) is known to exert anticancer and anti-metastatic effects on a variety of tumors. Recently, NC has also been shown to inhibit PIK3/AKT/mTOR axis in U87 human glioma cells.

METHODS

The study shows NC employing pDok2, caspase 3 dependent cell death in C6 rat glioma and U87 human malignant glioblastoma cells. The effect of NC on glioblastoma cell lines was accessed by MTT, clonogenic and wound healing assays. Cell cycle analysis was performed by FACS. Moreover, the effect of NC on downstream target proteins, such as caspase3, pDok2, PARP, and Gsk3 beta, were measured by western blotting.

RESULTS

Overexpressed pDok2 protein has recently been reported as a prognostic marker with poor outcomes for human glioblastoma multiformae. We found that NC inhibits pDok2 in U87 cells in a concentration-dependent way. We further showed that cleaved PARP and cleaved caspase 3 protein expressions were increased in C6 cells treated with NC in a dose-dependent way. NC effectively attenuated C6 cells growth and colony formation at 8μM (micromoles) concentration. Cell cycle arrest in G2/M phase was further confirmed by flow cytometry. NC also exhibited its inhibitory effect on Gsk3 beta, which has been proven to be altered in glioma biology.

CONCLUSIONS

Collectively, we predicted that NC could be employed as a potential anti-glioma mediator that needs attention to explore the mechanisms of its activity.

摘要

背景

盐酸去甲辛弗林（NC）已被证实对多种肿瘤具有抗癌和抗转移作用。最近，NC 还被证明可以抑制 U87 人胶质母细胞瘤细胞中的 PI3K/AKT/mTOR 轴。

方法

本研究表明，NC 通过 pDok2 诱导 C6 大鼠胶质瘤和 U87 人恶性神经胶质瘤细胞发生 caspase 3 依赖性细胞死亡。通过 MTT、集落形成和划痕愈合实验评估 NC 对神经胶质瘤细胞系的作用。通过 FACS 进行细胞周期分析。此外，通过 Western blot 测定 NC 对下游靶蛋白（如 caspase3、pDok2、PARP 和 Gsk3β）的影响。

结果

最近有报道称，过表达的 pDok2 蛋白是人多形性胶质母细胞瘤预后不良的一个预后标志物。我们发现 NC 以浓度依赖的方式抑制 U87 细胞中的 pDok2。我们进一步表明，NC 以剂量依赖的方式增加 C6 细胞中 cleaved PARP 和 cleaved caspase 3 蛋白的表达。NC 在 8μM（微摩尔）浓度下有效抑制 C6 细胞的生长和集落形成。通过流式细胞术进一步证实细胞周期阻滞在 G2/M 期。NC 还显示出对 Gsk3β 的抑制作用，Gsk3β 在神经胶质瘤生物学中已被证明发生改变。