MicroRNA-7 silencing EGFR/PI3K pathway reverses the malignant phenotype of U251 glioma cells / 肿瘤

ABSTRACT

Objective: To investigate the malignant phenotype of human glioma cells by microRNA-7 (miR-7) silencing epidermal growth factor receptor (EGFR)/ phosphatidylinositol kinase-3 (PI3K) pathway. Methods: The human U251 glioma cells were transfected with pri-miR-7. The expression of miR-7 was analyzed by real-time fluorogenic quantitative-PCR (RFQ-PCR). The expressions of epidermal growth factor receptor (EGFR), PI3K and AKT2 proteins were detected by immunocytochemistry and Western blotting. The cell growth curves were drawn, and the cell cycle distribution induced by miR-7 was determined by flow cytometry (FCM). The cell migration ability and tumorigenicity were detected by Transwell chamber assay and soft agar colony assay, respectively. Results: The result of RFQ-PCR showed that the expression level of miR-7 was up-regulated in human glioma cells transfected with pri-miR-7. The expression levels of EGFR, PI3K and AKT2 proteins were down-regulated in glioma U251 cells transfected with pri-miR-7 (P < 0.05). The cell proliferation rate was slowed down, and the proportion of S phase cells was reduced. The abilities of cell migration and soft agar colony formation of U251 cells transfected with pri-miR-7 were significantly reduced (P < 0.05). Conclusion: MiR-7 transfection can effectively silence the expressions of key members of EGFR/PI3K pathway and reverse the malignant phenotype of U251 cells and which is expected to become a new choice of glioma gene therapy. Copyright© 2011 by TUMOR.