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miR-133a增加喉癌细胞对顺铂化疗的敏感性及其对ATP7B表达的影响

发表时间:2013-06-15  浏览量:1110  下载量:531
全部作者: 王绪锐,王苹,赵晓东,戴晓天,祝威
作者单位: 吉林省肿瘤医院耳鼻咽喉头颈外科;吉林大学第一医院耳鼻咽喉头颈外科;吉林省人民医院耳鼻咽喉头颈外科
摘 要: 目的:探讨miR-133a对喉癌耐药细胞化疗敏感性的影响,了解miR-133a调控顺铂杀伤肿瘤细胞的分子机制。方法:采用miRNA poly加尾PCR的方法检测正常咽上皮细胞NP69,Hep-2和Hep-2/长春新碱(vincristine, VCR) miR-133a的水平;采用慢病毒重组载体pEZX-miR-133a转染Hep-2细胞和Hep-2/VCR耐药细胞株,细胞活力实验观察不同浓度顺铂对细胞存活的影响;通过免疫荧光染色和实时定量PCR方法检测pEZX-miR-133a转染细胞ATP7B的转录和蛋白表达变化。结果:Hep-2细胞和Hep-2/VCR表达ATP7B的荧光值明显高于NP-69细胞。顺铂50 μmol/L和100 μmol/L作用组转染pEZX-miR-133a的Hep-2/VCR,细胞存活明显低于转染pEZX-control的Hep-2/VCR细胞;miR-133a降低了Hep-2/VCR细胞ATP7B的转录和表达。结论:miR-133a能够增加喉癌细胞对顺铂的化疗敏感性,其机制可能与miR-133a下调ATP7B蛋白表达有关。
关 键 词: 耳鼻咽喉科学;喉癌;miRNA;化疗;ATP7B
Title: miR-133a increases the sensitivity of laryngeal cancer cells to cisplatin and affects expression of ATP7B
Author: WANG Xurui, WANG Ping, ZHAO Xiaodong, DAI Xiaotian, ZHU Wei
Organization: Department of Otorhinolaryngology, Cancer Hospital of Jilin Province; Department of Otorhinolaryngology, First Hospital of Jilin University; Department of Otorhinolaryngology, People’s Hospital of Jilin Province
Abstract: Objective: To investigate the influence of miR-133a on chemosensitivity of laryngeal carcinoma drug resistant cells, and clarify the molecular mechanisms of miR-133a regulation of cisplatin on killing tumor cells. Methods: miRNA were firstly poly tailed, and then applied PCR to detect miR-133a levels in normal pharyngeal epithelial cells NP69, Hep-2 and Hep-2/VCR. Then, lentiviral recombinant vector pEZX-miR-133a was transfected into Hep-2 cells and Hep-2/VCR cells, the effects of different concentrations of cisplatin on cell survival were observed by using cell viability experiments. The differences of ATP7B expression at transcription and protein level after being transfected pEZX-miR-133a were also detected by immunofluorescence staining and real-time PCR. Results: ATP7B fluorescence value was significantly higher in Hep-2 and Hep-2/VCR than in the NP-69 cells. The survival of pEZX-miR-133a Hep-2/VCR cells was significantly lower than the pEZX-control Hep-2/VCR cells after adding 50 μmol/L and 100 μmol/L cisplatin. In addition, miR-133a reduced transcription and expression of ATP7B in Hep-2/VCR cells. Conclusion: miR-133a could increase the chemosensitivity of laryngeal carcinoma cells to cisplatin, and the mechanism might be related to down-regulating of ATP7B protein expression by miR-133a.
Key words: otorhinolaryngology; laryngeal carcinoma; miRNA; chemotherapy; ATP7B
发表期数: 2013年6月第11期
引用格式: 王绪锐,王苹,赵晓东,等. miR-133a增加喉癌细胞对顺铂化疗的敏感性及其对ATP7B表达的影响[J]. 中国科技论文在线精品论文,2013,6(11):1033-1039.
 
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