受体酪氨酸激酶Axl对胶质母细胞瘤细胞增殖、凋亡和侵袭的影响

Effects of Receptor Tyrosine Kinase Axl on Proliferation, Apoptosis and Invasion of Glioblastoma Cells

目的:研究受体酪氨酸激酶Axl在胶质母细胞瘤组织和细胞系U-118MG细胞中的表达情况及其对U-118MG细胞增殖、凋亡、侵袭的影响。方法:收集2015年3月至2018年5月在本院进行手术切除并经病理分型证实的胶质母细胞瘤组织标本(n=30),另取脑外伤手术中因作内减压而切除的正常脑组织作为对照(n=28)。采用荧光实时定量(qRT-PCR)检测正常脑组织和胶质母细胞瘤肿瘤组织中Axl m RNA表达水平;采用Western blot检测人小神经胶质HM细胞、U-118MG细胞以及Axl-shRNA转染后U-118MG细胞中Axl蛋白表达水平;采用CCK-8检测Axl-shRNA转染后U-118MG细胞增殖能力;采用流式细胞术检测Axl-shRNA转染后U-118MG细胞凋亡水平;采用Transwell小室实验检测Axl-shRNA转染后U-118MG细胞的侵袭能力。结果:在胶质母细胞瘤组织中Axl m RNA表达水平显著高于正常脑组织(P<0.05);U-118MG细胞Axl蛋白表达水平显著高于人小神经胶质细胞系HM细胞,差异有统计学意义(P<0.05);转染Axl-shRNA后,U-118MG细胞中Axl蛋白表达水平显著降低(P<0.05)。与U-118MG细胞和转染control-shRNA细胞相比,转染Axl-shRNA的U-118MG细胞增殖能力降低(P<0.05),凋亡水平升高(P<0.05),侵袭能力降低(P<0.05)。结论:在胶质母细胞瘤组织和U-118MG细胞中,Axl表达水平显著增高,并且Axl表达水平与U-118MG细胞增殖、凋亡及侵袭密切关联。

Objective: To observe the expression of receptor tyrosine kinase Axl in glioblastoma tissues and U-118 MG cells, and the effects of Axl in the proliferation, apoptosis and invasion of U-118 MG cells. Methods: Glioblastoma tissue samples(n=30) which were surgically removed in our hospital and confirmed by pathological classification were collected from March 2015 to may 2018. In addition, the normal brain tissue which was removed due to internal decompression during brain trauma operation was taken as the control(n=28). The expression of Axl m RNA in normal brain tissue and glioblastoma tissue was detected by real time fluorescence quantitative PCR(qRT-PCR);the expression of Axl protein in human microglia HM cells, U-118 MG cells and U-118 MG cells after Axl shRNA transfection was detected by Western blot;the proliferation of U-118 MG cells after Axl shRNA transfection was detected by CCK-8;the proliferation of Axl shRNA was detected by flow cytometry. The apoptotic level of U-118 MG cells after transfection and the invasiveness of U-118 MG cells after Axl shRNA transfection were detected by Transwell cell experiment. Results: The expression of Axl m RNA in glioblastoma tissues were significantly higher than the normal brain tissues(P<0.05). The protein expression level of Axl in U-118 MG cells was significantly higher than the HM cells(P<0.05). Compared with control-shRNA group, Axl-shRNA down regulated the protein expression of Axl in U-118 MG cells(P<0.05). After transfected with Axl-shRNA, the proliferation and invasion ability of U-118 MG cells were reduced(P<0.05), and the cell apoptosis was increased(P<0.05). Conclusion: The expression levels of Axl in glioblastoma tissues and glioblastoma U-118 MG cells were increasd, which is closely related to the proliferation, apoptosis and invasion of glioblastoma cells.