摘要
目的探究MTA1基因敲除后对鼻咽癌细胞株5-8F细胞转移能力的影响。方法设计并构建针对MTA1基因的RNAi片段,脂质体(LipofectamineTM2000)介导Si-MTA1-01(Si-MTA1-01组)和Si-MTA1-02(Si-MTA1-02组)感染5-8F细胞,同时设无义序列转染组(Si-ctr组)为对照。应用荧光定量PCR和蛋白印迹法检测转染后各组细胞的MTAl mRNA和蛋白表达;细胞划痕实验、基底膜侵袭实验和细胞黏附实验检测转染后5-8F细胞迁移和侵袭能力的变化,并进行比较分析。结果与Si-ctr组相比,Si-MTA1-01和Si-MTA1-02组MTA1 mRNA及蛋白表达水平下降,穿膜细胞数减少,细胞黏附率增加(均P<0.05),划痕实验显示细胞阻滞效果明显。结论沉默MTA1能明显减弱鼻咽癌细胞的迁移和侵袭能力,MTA1有望成为治疗鼻咽癌的有效靶点。
Objective To investigate the effects of MTA1 knock down on migration and invasion of NPC cell 5-8F in vitro. Methods RNAi （Si-MTA1-01 and Si-MTA1-02） that can transiently silenced MTA1 was designed, synthesized and transfected into 5-8F cells by lipofectamine 2000. Control group （transfection with nonsense sequence） was also estab-lished. The efficiency of MTA1 depletion was determined by q-PCR and Western blot. Wound-healing assay ,Matrigel inva-sion assay and thesolid-phase adhesion assay were performed to investigate the effect of MTA1 knockdown on 5-8F cell me-tastasis. Results Transiently knock down of MTA1 decreased MTA1 transcription and expression in 5-8F cells compared to shRNA-con cells, showing by Real-time PCR and western blot. The invasion and migration of the cells transfected with siRNA-MTA1 were much weaker than the control group （P＆lt;0.05）. Conclusion silencing MTA 1 gene can effectively in-hibit the migration and invasion of nasopharyngeal carcinoma cell, and might be a promising target for NPC treatment.
出处
《天津医药》
CAS
北大核心
2014年第4期309-311,I0002,共4页
Tianjin Medical Journal
关键词
鼻咽肿瘤
RNA干扰
细胞迁移分析
细胞黏附
细胞系
肿瘤
印迹法
蛋白质
逆转录聚合酶链反
应
MTA1
鼻咽癌
5-8F细胞
nasopharyngeal neoplasms
RNA interference
cell migration assays
cell adhesion
cell line,tumor
blot-ting,Western
reverse transcriptase polymerase chain reaction
nasopharyngeal cancer
5-8F cell
