微丝相关蛋白1L2对胶质瘤细胞株LN229侵袭和迁移能力的影响  

Influence of actin filament associated protein 1-like 2 on invasion and migration of glioma cell line LN229

作　　者：孙树鹏[1] 王琼[2] 王修玉[1] 尚超[1] 王金环[2] 

机构地区：[1]天津医科大学研究生院,300070 [2] 天津市环湖医院 天津市脑血管与神经变性重点实验室

出　　处：《中华实验外科杂志》2015年第4期770-774,共5页Chinese Journal of Experimental Surgery

基　　金：天津市科委抗癌重大专项攻关计划资助项目（12ZCDZSY17700）;天津市科委重点项目（14JCZDJC35600）;天津市卫生局科技攻关计划资助项目（11KG115）

摘　　要：目的 观察下调胶质瘤细胞株LN229中微丝相关蛋白1L2 （AFAP1L2）的表达对细胞侵袭和迁移能力的影响.方法 将LN229细胞分为空白对照组、无义序列小于扰RNA （siRNA）转染组和AFAP1 L2 siRNA转染组进行细胞培养,细胞转染siRNA后,采用Transwell、划痕实验检测细胞侵袭和迁移能力;采用Western blot检测AFAP1L2、基质金属蛋白酶（MMP）-2、MMP-9的表达水平.结果 转染siRNA后,Western blot检测结果显示：空白对照组、无义序列siRNA转染组、AFAP1L2 siRNA转染组AFAP1L2蛋白相对表达量分别为1.00±0.10、1.00±0.13、0.52±0.06,差异有统计学意义（P＜0.05）;MMP-2蛋白相对表达量分别为1.00±0.12、0.92±0.08、0.21±0.02,MMP-9蛋白相对表达量分别为1.00±0.13、1.12±0.07、0.17±0.01,表明AFAP1 L2 siRNA转染组MMP-2和MMP-9蛋白表达降低（P＜0.05）.侵袭和迁移实验结果显示：空白对照组、无义序列siRNA转染组、AFAP1 L2 siRNA转染组Transwell侵袭实验穿膜细胞数分别为（246.74±10.52）、（236.85±11.74）、（40.14±8.32）个,Transwell迁移实验穿膜细胞数分别为（258.36±11.46）、（230.52±9.87）、（56.43±7.16）个,细胞划痕实验48 h迁移到空白处的细胞数分别为（296.60±27.14）、（113.80±25.43）、（51.20±13.25）个,说明转染AFAP1L2 siRNA组细胞侵袭和迁移能力明显低于其他两组（P＜0.05）.结论 沉默胶质瘤细胞中AFAP1L2的表达能显著抑制肿瘤细胞的侵袭和迁移能力.Objective To investigate the influence of actin filament associated protein 1-like 2 （AFAP1L2） on invasion and migration of glioma cell LN229.Methods LN229 cells were divided into the control group,nonsense sequence small interfering RNA （siRNA） transfected group and AFAP1L2 siRNA transfected group.The invasion and migration changes of LN229 cells transfected by siRNA were evaluated by transwell assay and scratch assay.After siRNAs were transfected into LN229 cells,the expression of AFAP1L2,matrix metalloproteinase （MMP）-2 and MMP-9 were detected by Western blotting.Results After transfection with siRNA,the results of Western blotting showed that the relative protein expression levels of AFAP1L2,MMP-2 and MMP-9 were 0.52 ±0.06,0.21 ±0.02 and 0.17 ±0.01 in the AFAP1L2 siRNA transfected group,1.00 ±0.10,1.00 ±0.12 and 1.00 ±0.13 in the control group,1.00 ±0.13,0.92 ±0.08 and 1.12 ±0.07 in the nonsense sequence group respectively （P 〈0.05）.The results of the transwell assay showed that the invasive and migrated membrane cell number were 40.14 ± 8.32 and 56.43 ±7.16 in the AFAP1L2 siRNA transfected group,246.74 ± 10.52 and 258.36 ± 11.46 in the control group,236.85 ± 11.74 and 230.52 ± 9.87 in the nonsense sequence group respectively （P 〈 0.05）.Results of wound healing assay in the control group,nonsense sequence group,AFAP1L2 siRNA transfected group were 296.60 ± 27.14,113.80 ± 25.43 and 51.20 ± 13.25 respectively （P 〈 0.05）.The transwell assay and scratch assay results showed that the ability of invasion and migration were significantly reduced in AFAP1 L2 siRNA transfected group （P 〈 0.05）.Conclusion Using RNAi technology,siRNA targeting AFAP1L2 efficiently knocks down the expression of AFAP1L2 in human glioma cells,results in suppression of tumor cell invasion and migration.AFAP1L2 is a potential therapeutic target for malignant gliomas.

关 键 词：微丝相关蛋白1L2 胶质瘤 侵袭 迁移 

分 类 号：R739.41[医药卫生—肿瘤]