天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2014年
4期
309-311,401
,共4页
涂英华%练祖平%谢有科
塗英華%練祖平%謝有科
도영화%련조평%사유과
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
鼻嚥腫瘤%RNA榦擾%細胞遷移分析%細胞黏附%細胞繫,腫瘤%印跡法,蛋白質%逆轉錄聚閤酶鏈反應%MTA1%鼻嚥癌%5-8F細胞
비인종류%RNA간우%세포천이분석%세포점부%세포계,종류%인적법,단백질%역전록취합매련반응%MTA1%비인암%5-8F세포
nasopharyngeal neoplasms%RNA interference%cell migration assays%cell adhesion%cell line,tumor%blot-ting,Western%reverse transcriptase polymerase chain reaction%MTA1%nasopharyngeal cancer%5-8F cell
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。
目的：探究MTA1基因敲除後對鼻嚥癌細胞株5-8F細胞轉移能力的影響。方法設計併構建針對MTA1基因的RNAi片段，脂質體（LipofectamineTM2000）介導Si-MTA1-01（Si-MTA1-01組）和Si-MTA1-02（Si-MTA1-02組）感染5-8F細胞，同時設無義序列轉染組（Si-ctr組）為對照。應用熒光定量PCR和蛋白印跡法檢測轉染後各組細胞的MTAl mRNA和蛋白錶達；細胞劃痕實驗、基底膜侵襲實驗和細胞黏附實驗檢測轉染後5-8F細胞遷移和侵襲能力的變化，併進行比較分析。結果與Si-ctr組相比，Si-MTA1-01和Si-MTA1-02組MTA1 mRNA及蛋白錶達水平下降，穿膜細胞數減少，細胞黏附率增加（均P<0.05），劃痕實驗顯示細胞阻滯效果明顯。結論沉默MTA1能明顯減弱鼻嚥癌細胞的遷移和侵襲能力，MTA1有望成為治療鼻嚥癌的有效靶點。
목적：탐구MTA1기인고제후대비인암세포주5-8F세포전이능력적영향。방법설계병구건침대MTA1기인적RNAi편단，지질체（LipofectamineTM2000）개도Si-MTA1-01（Si-MTA1-01조）화Si-MTA1-02（Si-MTA1-02조）감염5-8F세포，동시설무의서렬전염조（Si-ctr조）위대조。응용형광정량PCR화단백인적법검측전염후각조세포적MTAl mRNA화단백표체；세포화흔실험、기저막침습실험화세포점부실험검측전염후5-8F세포천이화침습능력적변화，병진행비교분석。결과여Si-ctr조상비，Si-MTA1-01화Si-MTA1-02조MTA1 mRNA급단백표체수평하강，천막세포수감소，세포점부솔증가（균P<0.05），화흔실험현시세포조체효과명현。결론침묵MTA1능명현감약비인암세포적천이화침습능력，MTA1유망성위치료비인암적유효파점。
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<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.
