天津医药

• 细胞与分子生物学 • 上一篇    下一篇

MTA1沉默抑制鼻咽癌侵袭能力的体外研究

涂英华   

  1. 广西中医药大学附属瑞康医院肿瘤科一区
  • 收稿日期:2013-10-14 修回日期:2013-12-12 出版日期:2014-04-15 发布日期:2014-04-15
  • 通讯作者: 涂英华

Silencing of MTA1Attenuates Nasopharyngeal Carcinoma Metastasis in vitro

TU ying hua   

  • Received:2013-10-14 Revised:2013-12-12 Published:2014-04-15 Online:2014-04-15
  • Contact: TU ying hua

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摘要：

【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（Lipofectamine TM 2000）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MTA1mRNA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。

关键词: MTA1基因, 鼻咽癌, 侵袭和转移, RNA干扰, 5-8F细胞

Abstract:

[Abstract] Objective To investigate the effects of MTA1knock down on migration and invasion of NPC cell5-8F in vitro. Methods RNAi (Si-MTA1-01and Si-MTA1-02) that can transiently silenced MTA1was designed, synthesized and transfected into5-8F cells by lipofectamine2000. Control group (transfection with nonsense sequence) was also estab? lished. The efficiency of MTA1depletion 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 MTA1knockdown on5-8F cell me? tastasis. Results Transiently knock down of MTA1decreased MTA1transcription 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-MTA1were much weaker than the control group (P<0.05). Conclusion silencing MTA1gene can effectively in? hibit the migration and invasion of nasopharyngeal carcinoma cell, and might be a promising target for NPC treatment.