Y Ye1, F Zhi, Y Peng, C-C Yang. 1. Department of Neurosurgery, The Third Affiliated Hospital of Soochow University, Changzhou, China. niis06@163.com.
Abstract
OBJECTIVE: The aim of this study was to explore whether miR-128 could promote the apoptosis of glioma cells by targeting NIMA related kinase-2 (NEK2), thus participating in the occurrence and progression of glioma. PATIENTS AND METHODS: Expression levels of miR-128 and NEK2 in glioma tissues and normal brain tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The relationship between miR-128 expression, tumor size and stage of glioma was analyzed. The effect of miR-128 on the apoptosis of glioma cells was detected by flow cytometry and Western blot, respectively. Dual-luciferase reporter gene assay was applied to verify the binding condition of miR-128 and NEK2. Meanwhile, rescue experiments were conducted to determine whether miR-128 could promote the apoptosis of glioma cells by targeting NEK2. RESULTS: The expression level of miR-128 in glioma tissues was significantly lower than that of normal brain tissues. However, NEK2 was highly expressed in glioma tissues. MiR-128 expression was correlated to tumor size and malignant level of glioma, whereas not related to age and gender of glioma patients. Meanwhile, overexpression of miR-128 promoted the apoptosis of U87 cells, upregulated protein levels of cleaved Caspase-3 and BCL2-associated X (Bax), and downregulated B-cell lymphoma-2 (Bcl-2). Dual-luciferase reporter gene assay indicated that miR-128 directly bound to NEK2. Further rescue experiments suggested that NEK2 overexpression partially reversed the effect of miR-128 on the apoptosis of glioma cells. CONCLUSIONS: Downregulated miR-128 inhibited the apoptosis of glioma cells via targeting NEK2.
OBJECTIVE: The aim of this study was to explore whether miR-128 could promote the apoptosis of glioma cells by targeting NIMA related kinase-2 (NEK2), thus participating in the occurrence and progression of glioma. PATIENTS AND METHODS: Expression levels of miR-128 and NEK2 in glioma tissues and normal brain tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The relationship between miR-128 expression, tumor size and stage of glioma was analyzed. The effect of miR-128 on the apoptosis of glioma cells was detected by flow cytometry and Western blot, respectively. Dual-luciferase reporter gene assay was applied to verify the binding condition of miR-128 and NEK2. Meanwhile, rescue experiments were conducted to determine whether miR-128 could promote the apoptosis of glioma cells by targeting NEK2. RESULTS: The expression level of miR-128 in glioma tissues was significantly lower than that of normal brain tissues. However, NEK2 was highly expressed in glioma tissues. MiR-128 expression was correlated to tumor size and malignant level of glioma, whereas not related to age and gender of gliomapatients. Meanwhile, overexpression of miR-128 promoted the apoptosis of U87 cells, upregulated protein levels of cleaved Caspase-3 and BCL2-associated X (Bax), and downregulated B-cell lymphoma-2 (Bcl-2). Dual-luciferase reporter gene assay indicated that miR-128 directly bound to NEK2. Further rescue experiments suggested that NEK2 overexpression partially reversed the effect of miR-128 on the apoptosis of glioma cells. CONCLUSIONS: Downregulated miR-128 inhibited the apoptosis of glioma cells via targeting NEK2.