Abstract. Aim: To investigate the effect of moesin expression on cell proliferaton and invasion of human glioblastoma cell lines in vitro. Materials and Methods: Glioblastoma LN229 and U87 cells were transfected with the H4645-plenti-EGFP-moesin expression vector for moesin up-regulation. Moesin and β-catenin expression levels in the transfectedGlioblastoma is a highly invasive recalcitrant cancer that is poorly understood (1-9). Moesin is a member of the ezrinradixin-moesin (ERM) protein family and is a link between the actin cytoskeleton and the plasma membrane. Moesin is involved in cell morphology, motility and adhesion, as well as other processes of tumorigenesis (10-12). Moesin has been studied in estrogen receptor (ER)-negative breast cancers (13), basal breast carcinomas (14), pancreatic cancer (15), melanoma (16), and gastric carcinoma (17). Moesin expression correlated with pathologic grade and poor clinical outcome, including survival in astrocytoma (18,19). However, the role of moesin in human glioblastoma progression is incompletely understood and, thus, the topic of the present report.In this study, we investigated moesin's effect on the growth and invasion ability of the glioblastoma cells by upregulation of the gene (MSN).
Materials and MethodsCell culture. The glioblastoma cell line LN229 and U87 were purchased from the Shanghai cell bank of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in RPMI 1640 medium (GIBCO Life Technologies, Grand Island, NY, USA) containing 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA), 100 U/ml penicillin and 100 μg/ml streptomycin (Thermo Fisher Scientific) at 37˚C with 5% CO 2 .Moesin-expression vector transfection. The H4645-plenti-enhanced green fluorescent protein (EGFP)-moesin expression vector was used for up-regulation of the moesin gene (MSN) in the LN229 and U87 glioblastoma cells (LN229-H4645 and U87-H4645, respectively). The H149 plenti-EGFP empty vector was used as negative control (LN229-H149 and U87-H149, respectively). The moesin expression vector was constructed by Obio Technology Corp., Ltd. (Shanghai, China). Cultured glioblastoma LN229 and U87 cells were transfected with the moesin expression vector according to the manufacturer's instructions. After 24h transfection, the medium was changed. Cells were then observed under fluorescence microscopy at 48 hours after transfection. Cells were selected and treated with 500-800 μg/ml G148 (Micropoly Biotech Co., Ltd, Nanjing, China) for 1 week. Images were obtained from fluorescence microscopy. Transfection efficiency of the expression vector in the two cell lines was 2211 This article is freely accessible online.