去整合素样金属基质酶8对脑胶质瘤增殖和侵袭的影响及其机制  

The influence of A disintegrin and metalloprotease protein 8 in glioma progression and its mechanism

作　　者：郭松波 孙剑瑞[1] 张所军 Guo Songbo;Sun Jianrui;Zhang Suojun(Department of Neurosurgery,the First Afiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Neurosurgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]郑州大学第一附属医院神经外科,郑州450052 [2]华中科技大学同济医学院附属同济医院神经外科,武汉430030

出　　处：《中华实验外科杂志》2024年第10期2248-2251,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金面上项目(82072805)。

摘　　要：目的:探讨去整合素样金属基质酶8(ADAM8)对巨噬细胞迁移和血管生成的影响及对胶质瘤细胞侵袭的作用及其机制。方法:骨髓细胞分别取自ADAM8野生型和ADAM8基因敲除C57Bl/6小鼠腿骨骨髓。骨髓细胞分化为巨噬细胞备用。采用划痕实验比较巨噬细胞ADAM8野生型和ADAM8基因敲除细胞迁移能力;并比较两种类型巨噬细胞上清液对HUVEC细胞血管生成的影响,采用聚合酶链反应(PCR)实验及PrAMA实验检测ADAM8基因敲除对其他金属基质酶的mRNA表达及酶活性影响。应用Transwell侵袭实验比较巨噬细胞ADAM8对胶质瘤细胞GL261侵袭能力的作用。组间数据比较采用非配对t检验。结果:ADAM8能够促进巨噬细胞的迁移能力,DMEM两组野生型细胞迁移能力(1.00±0.04)高于ADAM8基因敲除组(0.72±0.05),差异有统计学意义(t=3.200,P<0.01),LPS组野生型细胞迁移能力(1.30±0.09)高于ADAM8基因敲除组(0.92±0.07),差异有统计学意义(t=4.132,P<0.01),ADAM8显著促进野生型组的血管生成,野生型组生成小管结数量(13.84±4.82)高于ADAM8基因敲除组(4.39±1.83),差异有统计学意义(t=2.170,P<0.05)。ADAM8不影响巨噬细胞中MMP-9的mRNA表达,但ADAM8的敲除降低了MMP-9的活性,野生型MMP-9的活性比ADAM8敲除巨噬细胞高2倍以上。同时发现ADAM8促进了胶质瘤细胞GL261的侵袭,野生型巨噬细胞组GL261细胞穿透基质膜数量(187.67±23.23)高于ADAM8基因敲除组(2.67±1.69),差异有统计学意义(t=11.230,P<0.01)。结论:ADAM8影响巨噬细胞的迁移能力,并影响胶质瘤的血管生成和侵袭。Objective:To investigate the effects of A disintegrin and metalloprotease protein 8(ADAM8)on the migration and angiogenesis of macrophages and the invasion of glioma cells.Methods:The marrow cells were harvested from the femur and tibia bones of the C57Bl/6 wild-type mice and ADAM8 deficient mice,then differentiate to the macrophages.Wound healing assay was used to compare the migration ability of ADAM8 wild-type macrophages and ADAM8 knockout macrophages,and the effects of the supernatants of the two types of macrophages on the angiogenesis of HUVEC were compared,and the effects of ADAM8 knockout on the mRNA expression and catalytic activity of other matrix metalloproteinases were detected by polymerase chain reaction(PCR)and PrAMA assay.Transwell invasion assay was used to compare the effect of two types of macrophage supernatants on the invasion ability of glioma cells GL261.The statistical significance was evaluated by unpaired student’s t-test.Results:ADAM8 could promote the migration.Either in DMEM medium or with the LPS stimulation the knockout of Adam8 on macrophages decreased cell migration significantly(0.72±0.05,0.92±0.07)compared to wild-type macrophages(1.00±0.04,1.30±0.09,t=3.200,4.132,P<0.01).ADAM8 could promote the angiogenesis significantly in the wild-type group(13.84±4.82),compared to the A8_KO group(4.39±1.83,t=2.170,P<0.05).ADAM8 did not affect MMP-9 mRNA expression in macrophages,but knockout of ADAM8 decreased MMP-9 activity more than two times.It was also found that ADAM8 promoted the invasion of glioma cells GL261.There were more GL261 cells infiltrate the Matrigel in the wild-type group(187.67±23.23)than the ADAM8 knockout group(2.67±1.69,t=11.230,P<0.01).Conclusion:ADAM8 affects the invasive ability of macrophages and promote the angiogenesis and invasion of glioma.

关 键 词：胶质瘤 去整合素样金属基质酶8 巨噬细胞 肿瘤微环境 

分 类 号：R739.41[医药卫生—肿瘤]