摘要
目的观察塞来昔布（celecoxib）与顺铂（DDP）联合处理对DDP耐药的鼻咽癌TW03／DDP细胞生长的影响。方法用（0．05-1．00）μg／mL的DDP梯度培养TW03／DDP细胞，经反转录PCR和Western blot法证明TW03／DDP细胞肺耐药相关蛋白（LRP）高表达后，采用siRNA干扰TW03／DDP细胞LRP蛋白表达；Celecoxib与DDP联合或单独处理不具耐药性的鼻咽癌TW03细胞、TW03／DDP细胞和TW03／DDP—siRNA细胞。CCK-8法检测细胞的生长抑制率，annexin V-FITC／碘化丙锭（PI）双染色结合流式细胞术检测细胞凋亡率。结果Celecoxib与DDP单独使用均能抑制TW03／DDP细胞和TW03细胞的生长并诱导凋亡，但DDP对TW03／DDP细胞作用弱；Celecoxib与DDP联用对2个细胞株的生长都有抑制作用，其生长抑制率和凋亡率均大于二者单独应用的效果；Celecoxib处理TW03／DDP细胞可使其LRP的mRNA和蛋白水平下降。结论Celecoxib和DDP联合应用可抑制LRP的表达，显著抑制TW03／DDP细胞的生长并诱导其凋亡。
Objective To investigate the synergistic cytotoxicity of cis-dichlorodiamine platinum （DDP） and celecoxib on nasopharyngeal carcinoma TW03 cell which is resistant to DDP （TW03/DDP）. Methods The mRNA and protein levels of lung resistance-related protein （LRP） were detected by reverse transcription PCR and Western blotting in TW03/DDP cells after （0.05-1.00） μg/mL DDP treatment. TW03/DDP clone which highly expressed LRP was transfected with siRNA targeting LRP mRNA. Nasopharyngeal carcinoma TW03 cells, TW03/DDP cells and TW03/DDP-siRNA cells were treated respectively with DDP or celecoxib or combination of the two. CCK-8 assay was performed for detecting proliferation. Annexin V-FITC/PI staining combined with flow cytometry were used for detecting apoptosis rate. Results Both DDP and celecoxib treatment inhibited the proliferation and induced the apoptosis of TW03 cells and TW03/DDP cells, but only celecoxib showed a significant effect on TW03/DDP cells, but not DDP. Combination of DDP and celecoxib showed a synergistic role in inducing the apoptosis of both TW03 cells and TW03/DDP cells. Meanwhile, LRP mRNA and protein levels were down-regulated in celecoxib-treated TW03/DDP cells. Conclusion DDP and celecoxib showed a synergistic effect on proliferation inhibition and apoptosis induction of TW03/DDP cells by clown-regulating LRP mRNA and protein levels.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2015年第2期203-206,211,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
四川省科技厅科技支撑计划(2012SZZ011)
关键词
塞来昔布
顺铂
鼻炎肿瘤
肺耐药相关蛋白
耐药性
RNA干扰
celecoxib
DDP
TW03 nasopharyngeal cell line
lung resistance-related protein
drug resistance
RNAinterference
