Original Article
miR-17 enhances proliferation and migration and inhibits apoptosis in glioma cells by regulating SASH1 expression
Abstract
Background: To investigate the mechanisms underlying the regulation of human glioma cell growth, proliferation, and apoptosis by microRNA-17 (miR-17) to provide targets for novel human glioma therapies.
Methods: The expression of miR-17 in human glioma tissues and cell lines was detected using real-time quantitative PCR (qRT-PCR). A miR-17 inhibitor was transfected into the U251 human glioma cell line using liposomes, and changes in miR-17 expression were detected using qRT-PCR. The effects of miR-17 on numerous biological characteristics of U251 cells, including viability, proliferation, apoptosis, and migration, were analyzed using MTT, flow cytometry, and Transwell migration chamber assays. A luciferase reporter gene system was used to validate SAM- and SH3-domain containing 1 (SASH1) as a true target of miR-17. The effects of miR-17 on SASH1 protein expression were determined using western blotting.
Results: qRT-PCR results showed that compared with adjacent tissues, miR-17 expression was significantly increased in human glioma tissues and cells lines (P<0.001). Compared with the control group, miR-17 expression in the miR-17 inhibitor transfection group was significantly decreased (P<0.01). The results of the MTT, flow cytometry, and Transwell migration chamber assays showed that cell viability (P<0.05), G2M+S phase fraction (P<0.05), and migration ability (P<0.05) in the miR-17 inhibitor transfection group was significantly decreased compared with the normal control and inhibitor control groups. Flow cytometry showed that apoptosis in the miR-17 inhibitor transfection group was increased compared with the normal control and inhibitor control groups (P<0.05). Relative luciferase activity in the miR-17 mimic group was significantly decreased compared with the control group (P<0.05). Western blotting revealed that SASH1 protein expression in the miR-17 inhibitor transfection group was significantly increased compared with the normal control and inhibitor control groups (P<0.05).
Conclusions: miR-17 promotes viability, proliferation, and migration and inhibits apoptosis in U251 glioma cells, and these mechanisms could be mediated by the regulation of SASH1 gene expression.
Methods: The expression of miR-17 in human glioma tissues and cell lines was detected using real-time quantitative PCR (qRT-PCR). A miR-17 inhibitor was transfected into the U251 human glioma cell line using liposomes, and changes in miR-17 expression were detected using qRT-PCR. The effects of miR-17 on numerous biological characteristics of U251 cells, including viability, proliferation, apoptosis, and migration, were analyzed using MTT, flow cytometry, and Transwell migration chamber assays. A luciferase reporter gene system was used to validate SAM- and SH3-domain containing 1 (SASH1) as a true target of miR-17. The effects of miR-17 on SASH1 protein expression were determined using western blotting.
Results: qRT-PCR results showed that compared with adjacent tissues, miR-17 expression was significantly increased in human glioma tissues and cells lines (P<0.001). Compared with the control group, miR-17 expression in the miR-17 inhibitor transfection group was significantly decreased (P<0.01). The results of the MTT, flow cytometry, and Transwell migration chamber assays showed that cell viability (P<0.05), G2M+S phase fraction (P<0.05), and migration ability (P<0.05) in the miR-17 inhibitor transfection group was significantly decreased compared with the normal control and inhibitor control groups. Flow cytometry showed that apoptosis in the miR-17 inhibitor transfection group was increased compared with the normal control and inhibitor control groups (P<0.05). Relative luciferase activity in the miR-17 mimic group was significantly decreased compared with the control group (P<0.05). Western blotting revealed that SASH1 protein expression in the miR-17 inhibitor transfection group was significantly increased compared with the normal control and inhibitor control groups (P<0.05).
Conclusions: miR-17 promotes viability, proliferation, and migration and inhibits apoptosis in U251 glioma cells, and these mechanisms could be mediated by the regulation of SASH1 gene expression.
