干扰核转录因子Snail增强喉癌耐药细胞顺铂敏感性的研究  

Small interference of transcription factor Snail contribute to enhanced cisplatin sensitivity on human laryngeal resistant cancer cells

作　　者：于锋[1] 龚小蓉 周毅波[1] 刘炜 YU Feng GONG Xiaorong ZHOU Yibo LIU Wei(Institute of Otolaryngology Head and Neck Surgery, Affiliated to Guangzhou Medical University, Guangzhou Hospital of Otolaryngology Head and Neck Surgery, Guangzhou 12th People＇s Hospital,Guangzhou, 510620, Chin)

机构地区：[1]广州医科大学耳鼻咽喉头颈外科研究所广州市耳鼻咽喉头颈外科医院广州市第十二人民医院,广州510620

出　　处：《临床耳鼻咽喉头颈外科杂志》2018年第5期350-354,共5页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：广州市喉癌临床医学与转化中心试点建设项目(No:201604020005)

摘　　要：目的:探讨核转录因子Snail对喉癌顺铂耐药细胞株(Hep-2/CDDP)顺铂敏感性的影响。方法:采用小干扰RNA(si-RNA)技术靶向沉默Hep-2/CDDP细胞中Snail的表达;采用实时定量荧光聚合酶链式反应(RT-qPCR)法检测其干扰效率;免疫荧光法检测干扰组(si-Hep-2/CDDP)和对照组(Hep-2/CDDP)中Snail蛋白的表达情况;CCK-8法检测干扰组和对照组对0、1、2、4、8、16μg/ml不同浓度的顺铂抑制率;Western blot法检测干扰组和对照组si-Snail处理前后Snail、E-cadherin、MDR1蛋白的表达情况。结果:RT-qPCR结果显示,靶向沉默Snail后干扰组细胞Snail表达较对照组下降(67.85±9.50)%;免疫荧光实验示,干扰组细胞核及细胞质中Snail荧光强度均下降;CCK-8实验示,不同浓度顺铂作用后,与耐药细胞Hep-2/CDDP相比,干扰组抑制率较对照组升高;Western blot示Snail干扰组蛋白水平较耐药细胞组表达下调(P<0.05),同时上皮表型标记E-cadherin表达上调,多药耐药蛋白MDR1表达下调(均P<0.05)。结论:干扰核转录因子Snail可上调E-cadherin的表达,降低耐药细胞Hep-2/CDDP耐药蛋白MDR1的表达,增强喉癌耐药细胞的顺铂敏感性。Objective：To study the relationship between transcription factor Snail and the sensitivity of cisplatin on human laryngeal resistant cancer cells.Method：siRNA interference of Snail was transfected by small RNA interference technology.The interference efficiency on mRNA level were detected by RT-qPCR assay;the expression of Snail protein level was assessed by immunofluorescence.The inhibition ratio of different cisplatin concentration（0,1,2,4,8,16μg/ml）was detected by CCK-8 assay;the protein level of Snail,E-cadherin,MDR1 were detected by Western blot assay.Result：RT-qPCR assay show the expression of Snail on mRNA level was decreased to（67.85±9.50）% after transfection in Hep-2/CDDP cell（P〈0.05）.Immunofluorescence show fluorescence intensity of si-Hep-2/CDDP group was reduced both in nucleus and cytoplasm;CCK-8 assay show the inhibitory ratio of transfected group was increased compared to negative control and Hep-2/CDDP group in different cisplatin concentration（0,1,2,4,8,16μg/ml）（P〈0.05）.Western blot assay show the protein expression of Snail and MDR1 were down-regulated in transfected Hep-2/CDDP cells（all P〈0.05）,while epithelial marker E-cadherin was up-regulated in protein level（P〈0.05）.Conclusion：Small interference of transcription factor Snail could increase the expression of E-cadherin while decrease the expression of MDR1,and it was confirmed that interference Snail contribute to enhanced cisplatin sensitivity on human laryngeal resistant cancer cells.

关 键 词：喉肿瘤 SNAIL 耐药 顺铂 小干扰RNA 化疗敏感性 

分 类 号：R739.6[医药卫生—肿瘤]