Fan Wang1, Fenglong Chen1, Weipeng Hu2, Yi Zhang1. 1. Department of Neurosurgery, Third Hospital of Xiamen, Xiamen 361100, China. 2. Department of Neurosurgery, Second Affiliated Hospital of Fujian Medical University, Quanzhou 362000, China.
Abstract
OBJECTIVE: To investigate the inhibitory effects of silencing migration-inducing gene-7 (Mig-7) on vasculogenic mimicry formation, migration and invasion of human glioma cells in vitro and whether MEK/ERK signaling pathway mediates these effects. METHODS: Human glioma U251 cells were infected by lentiviral vectors carrying a small interfering RNA targeting Mig-7 gene (sh-Mig-7) or a negative control shRNA (sh-NC), and real-time quantitative PCR was used to detect the expression level of Mig-7 mRNA in the cells. Three-dimensional culture and Transwell chamber invasion assay were used to observe the effect of Mig-7 gene silencing on vasculogenic mimicry formation and invasion ability of the U251 cells. Western blotting was performed to detect the changes in the protein expression levels of MEK/ERK in the infected cells. RESULTS: We successfully obtained a U251 cell line with stable low expression of Mig-7 gene using RNA interference technique. Compared with the cells infected with sh-NC lentivirus and the non- infected cells, U251 cells infected with the lentiviral vector carrying sh-Mig-7 showed significantly decreased expression level of Mig-7 (P < 0.01) with obviously lowered vasculogenic mimicry formation and invasion abilities (P < 0.05). Mig-7 silencing also significantly lowered the expressions of MEK and ERK proteins in U251 cells (P < 0.05). CONCLUSIONS: Silencing of Mig-7 gene inhibits vasculogenic mimicry formation and invasion of U251 cells possibly by suppressing MEK/ERK signaling, suggesting the important role of Mig-7 gene in vasculogenic mimicry formation and invasion of human glioma cells.
OBJECTIVE: To investigate the inhibitory effects of silencing migration-inducing gene-7 (Mig-7) on vasculogenic mimicry formation, migration and invasion of humanglioma cells in vitro and whether MEK/ERK signaling pathway mediates these effects. METHODS:Humanglioma U251 cells were infected by lentiviral vectors carrying a small interfering RNA targeting Mig-7 gene (sh-Mig-7) or a negative control shRNA (sh-NC), and real-time quantitative PCR was used to detect the expression level of Mig-7 mRNA in the cells. Three-dimensional culture and Transwell chamber invasion assay were used to observe the effect of Mig-7 gene silencing on vasculogenic mimicry formation and invasion ability of the U251 cells. Western blotting was performed to detect the changes in the protein expression levels of MEK/ERK in the infected cells. RESULTS: We successfully obtained a U251 cell line with stable low expression of Mig-7 gene using RNA interference technique. Compared with the cells infected with sh-NC lentivirus and the non- infected cells, U251 cells infected with the lentiviral vector carrying sh-Mig-7 showed significantly decreased expression level of Mig-7 (P < 0.01) with obviously lowered vasculogenic mimicry formation and invasion abilities (P < 0.05). Mig-7 silencing also significantly lowered the expressions of MEK and ERK proteins in U251 cells (P < 0.05). CONCLUSIONS: Silencing of Mig-7 gene inhibits vasculogenic mimicry formation and invasion of U251 cells possibly by suppressing MEK/ERK signaling, suggesting the important role of Mig-7 gene in vasculogenic mimicry formation and invasion of humanglioma cells.
Authors: Jason E Fish; Manuel Cantu Gutierrez; Lan T Dang; Nadiya Khyzha; Zhiqi Chen; Shawn Veitch; Henry S Cheng; Melvin Khor; Lina Antounians; Makon-Sébastien Njock; Emilie Boudreau; Alexander M Herman; Alexander M Rhyner; Oscar E Ruiz; George T Eisenhoffer; Alejandra Medina-Rivera; Michael D Wilson; Joshua D Wythe Journal: Development Date: 2017-05-23 Impact factor: 6.868
Authors: Dongsic Choi; Laura Montermini; Dae-Kyum Kim; Brian Meehan; Frederick P Roth; Janusz Rak Journal: Mol Cell Proteomics Date: 2018-07-13 Impact factor: 5.911
Authors: Artur Czekierdowski; Sylwia Czekierdowska; Norbert Stachowicz; Tomasz Łoziński; Grzegorz Gurynowicz Journal: Ginekol Pol Date: 2017 Impact factor: 1.232
Authors: James A McCubrey; Linda S Steelman; William H Chappell; Stephen L Abrams; Richard A Franklin; Giuseppe Montalto; Melchiorre Cervello; Massimo Libra; Saverio Candido; Grazia Malaponte; Maria C Mazzarino; Paolo Fagone; Ferdinando Nicoletti; Jörg Bäsecke; Sanja Mijatovic; Danijela Maksimovic-Ivanic; Michele Milella; Agostino Tafuri; Francesca Chiarini; Camilla Evangelisti; Lucio Cocco; Alberto M Martelli Journal: Oncotarget Date: 2012-10