Sinomenine Hydrochloride Inhibits the Metastasis of Human Glioblastoma Cells by Suppressing the Expression of Matrix Metalloproteinase-2/-9 and Reversing the Endogenous and Exogenous Epithelial-Mesenchymal Transition.

Yumao Jiang,Yue Jiao,Tao Li,Yujuan Li,Yang Liu,Danqiao Wang,Xiaoliang Zhao,Meiyu Zhang,Zhiguo Wang

doi:10.3390/ijms19030844

Abstract

As shown in our previous study, sinomenine hydrochloride (SH), the major bioactive alkaloid isolated from Sinomenium acutum Rehd. et Wils. (Fam. Menispermaceae), initiates the autophagy-mediated death of human glioblastoma cells by generating reactive oxygen species and activating the autophagy-lysosome pathway. However, its effects on the migration and invasion of human glioblastoma cells have not yet been elucidated. Therefore, human glioblastoma U87 and SF767 cells were treated with SH (0.125 and 0.25 mM) for 24 h, and cell migration and invasion were assessed using scratch wound healing, migration and invasion assays. SH promoted G0/G1 phase arrest, inhibited the migration and invasion of the two cell lines, suppressed the activation of nuclear factor kappa B (NFκB) and the expression of matrix metalloproteinase (MMP)-2/-9, triggered endoplasmic reticulum (ER) stress, reversed the exogenous epithelial-mesenchymal transition (EMT) induced by the inflammatory microenvironment and the endogenous EMT. Additionally, NFκB p65 overexpression blocked the SH-mediated inhibitory effects on MMP-2/-9 expression and cell invasion. SH-induced autophagy was reduced in CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) or autophagy-related 5 (ATG5)-silenced human glioblastoma cells and cells treated with 4-phenylbutyric acid (4-PBA) or 3-methyladenine (3-MA), as shown by the decreased levels of the microtubule-associated protein light chain 3B (LC3B)-II and autophagic vacuoles (AVs) stained with monodansylcadaverine (MDC), respectively. Moreover, knockdown of CHOP or ATG5 and treatment with 4-PBA or 3-MA abolished the SH-mediated inhibition of mesenchymal markers (vimentin, Snail and Slug) expression and cell invasion, respectively. Importantly, SH also regulated the above related pathways in nude mice. Based on these findings, SH inhibited cell proliferation by inducing cell cycle arrest, and attenuated the metastasis of U87 and SF767 cells by suppressing MMP-2/-9 expression and reversing the endogenous and exogenous EMT in vitro and/or in vivo. Thus, SH might be a new potential anti-metastasis agent for the treatment of human glioblastoma.

Highlights

Glioblastoma, a most common primary tumor of the central nervous system, shows substantial migration and invasion, resulting in frequent metastases into the surrounding tissues [1]
Sinomenine has recently been reported to inhibit NFκB activation [9] or suppress matrix metalloproteinase (MMP)-2/-9 expression [10] to abrogate the invasiveness of various types of cancer cells, researchers have not determined whether sinomenine hydrochloride (SH) suppresses human glioblastoma cell metastasis through related mechanisms
Our results revealed that SH inhibited proliferation by inducing cell cycle arrest and attenuated the metastasis of human glioblastoma U87 and SF767 cells by suppressing the expression of MMP-2/-9 and reversing endogenous and exogenous epithelial-mesenchymal transition (EMT) in vitro and/or in vivo

Summary

Introduction

Glioblastoma, a most common primary tumor of the central nervous system, shows substantial migration and invasion, resulting in frequent metastases into the surrounding tissues [1]. The identification of potentially novel therapeutic agents that effectively inhibit human glioblastoma cell metastasis is urgently needed. Matrix metalloproteinases (MMPs) play a central role in the invasion process by degrading many elements of the extracellular matrix (ECM), including collagens, fibronectin and laminins, and suppression of MMP-2/-9 expression represents a potential strategy for preventing tumor cell invasion [3,4]. Phorbol 12-myristate 13-acetate-induced MMP-9 expression is regulated by nuclear factor kappa B (NFκB) in human breast cancer cells [5]. Sinomenine has recently been reported to inhibit NFκB activation [9] or suppress MMP-2/-9 expression [10] to abrogate the invasiveness of various types of cancer cells, researchers have not determined whether sinomenine hydrochloride (SH) suppresses human glioblastoma cell metastasis through related mechanisms

Objectives

Methods

Results