Gliomas
Effect of silencing abelson nonreceptor tyrosine kinases 2 expression on invasion and migration of glioma cells
Lei Chen, Meng Zhu, Shengping Yu, Chen Zhang, Pengfei Zhao, Hua Zhou, Long Hai, Bo Liu, Shuai Li, Xingchen Zhou, Yu Lin, Xuejun Yang
Published 2015-03-15
Cite as Chin J Neuromed, 2015, 14(3): 221-226. DOI: 10.3760/cma.j.issn.1671-8925.2015.03.002
Abstract
ObjectiveTo investigate the inhibitory effect of abelson nonreceptor tyrosine kinases 2 (ABL2) silenced by shRNA on invasion and migration in glioma cells as well as its potential mechanism.
MethodsThe ABL2 expressions in normal brain tissues and glioblastoma tissues, collected from patients performed epilepsy and glioma surgeries, respectively, in our hospital from September 2012 to August 2014, were detected by immunohistochemistry. The recombinant lentivirus aimed at ABL2 silencing was prepared. Human glioma cells SNB19 were employed in this study and assigned into three groups: group of ABL2 silenced by shRNA (shRNA-ABL2), shRNA-negative control group (shRNA-NC) and non-silencing shRNA group (shRNA-N). Forty-eight h after the transfection, the following experiments were performed: Real time quantitative-PCR was employed to detect the mRNA expression of ABL2; and Western blotting was used to detect the protein expression of ABL2, cortactin and phosphorylated cortactin at tyrosine 421 (pY-421cortactin), respectively; migration and invasion ability were evaluated by wound-healing assay and Transwell assay; immunofluorescence was used to disclose the expression relationship between ABL2 and cortactin at protein level.
ResultsImmunohistochemistry indicated that the ABL2 expression level in glioblastoma tissues was higher as compared with that in normal brain tissues. The mRNA and protein expression levels of ABL2 in shRNA-ABL2 group were significantly lower than those in the shRNA-NC group and shRNA-N group (P<0.05). Wound-healing assay indicated that the number of cell migration in the shRNA-ABL2 group (72.33±7.64) was significantly smaller than that in the shRNA-NC group and shRNA-N group (187.67±5.03 and 190.33±7.23, P<0.05); Transwell assay showed that the invasiveness capability of cells in the shRNA-ABL2 group was significantly decreased as compared with that in the shRNA-NC group and shRNA-N group (P<0.05). Double immunofluorescent staining revealed ABL2 enjoyed co-localization with cortactin at cell protruded membrane.
ConclusionsThe ABL2 expressions in glioblastoma tissues are higher than those in normal brain tissues. Silencing ABL2 expression will inhibit glioma cell migration and invasion owing to down-regulating the level of phosphorylated form of cortactin.
Key words:
Glioma; Abelson nonreceptor tyrosine kinases 2; Cortactin; Migration and invasion
Contributor Information
Lei Chen
Department of Neurosurgery, General Hospital of Tianjin Medical University, Tianjin 300052, China
Meng Zhu
Shengping Yu
Chen Zhang
Pengfei Zhao
Hua Zhou
Long Hai
Bo Liu
Shuai Li
Xingchen Zhou
Yu Lin
Xuejun Yang
