鼻咽癌多药耐药基因的表达及药物逆转的相关研究  

Expression of multidrug resistance gene in nasopharyngeal carcinoma and drug reversal

作　　者：徐洪波[1] 何斌[1] 孙谦[1] 张亚军[1] 蔡丰[1] 万强琨 汪庚明[1] XU Hong-bo(Department of cancer radiotherapy,the first affiliated hospital of Bengbu Medical College,Bengbu,Anhui,233004,China.)

机构地区：[1]蚌埠医学院第一附属医院肿瘤放射治疗科,安徽蚌埠233004

出　　处：《齐齐哈尔医学院学报》2020年第9期1061-1064,共4页Journal of Qiqihar Medical University

基　　金：蚌埠医学院自然科学基金重点项目(BYKY1732ZD)。

摘　　要：目的探讨鼻咽癌多药耐药基因的表达及药物逆转机制。方法取人鼻咽低分化鳞癌细胞系HNE1,用新辅助化疗药奈达铂(NDP)作为体外诱导剂,建立耐NDP的人鼻咽癌细胞系HNE1/NDP。qRT-PCR和免疫印迹检测HNE1和HNE1/NDP中多药耐药基因1(MDR1)、多药耐药相关蛋白1(MRP1)和P-糖蛋白(P-gp)的mRNA和蛋白表达差异;不同浓度的贝伐单抗(5、10、20、40、80μg/ml)处理HNE1和HNE1/NDP,48 h后CCK8法检测贝伐单抗对HNE1和HNE1/NDP细胞的增殖作用。不同浓度贝伐单抗联合1 ug/ml奈达铂处理HNE1/NDP后,CCK8检测贝伐单抗对HNE1/NDP的耐药逆转作用。流式细胞仪检测贝伐单抗处理HNE1/NDP细胞后细胞凋亡率。结果qRT-PCR和免疫印迹结果显示HNE1/NDP中MDR1、MRP1和P-gp的mRNA和蛋白表达显著上调;贝伐单抗呈浓度依赖性地抑制HNE1和HNE1/NDP细胞的增殖,同时逆转HNE1/NDP的耐药性;qRT-PCR结果显示贝伐单抗处理HNE1/NDP细胞后,抑制MDR1、MRP1和P-gp的mRNA表达。PI染色结果显示贝伐单抗促进HNE1/NDP细胞的凋亡。结论贝伐单抗通过下调多药耐药相关基因的表达,同时促进HNE1/NDP细胞凋亡,从而实现耐药逆转作用。Objective To explore the expression of multidrug resistance gene in nasopharyngeal carcinoma and drug reversal mechanism.Methods The human nasopharyngeal poorly differentiated squamous cell carcinoma cell line HNE1 was obtained,and the neoadjuvant drug Nidaplatin(NDP)was used as an inducing agent to establish a human NDP-resistant NDP cell line HNE1/NDP.mRNA and protein expression of multidrug resistance gene 1(MDR1),multidrug resistance associated protein 1(MRP1)and P-glycoprotein(P-gp)in HNE1 and HNE1/NDP cell lines by qRT-PCR and western blotting.HNE1 and HNE1/NDP cells were treated with different concentrations of bevacizumab(5,10,20,40,80μg/ml)for 48 h,and CCK8 was used to detect the proliferation of HNE1 and HNE1/NDP cells.After treatment with different concentrations of bevacizumab combined with 1ug/ml nedaplatin for HNE1/NDP,CCK8 detected the reversal effect of bevacizumab on HNE1/NDP.Flow cytometry was used to detect the apoptosis rate of HNE1/NDP cells after treatment of bevacizumab.Results qRT-PCR and western blotting results showed that mRNA and protein expression of MDR1,MRP1 and P-gp were significantly up-regulated in HNE1/NDP cell lines.Bevacizumab inhibited the proliferation activity of HNE1 and HNE1/NDP cells in a concentration-dependent manner,and reversed the drug resistance of HNE1/NDP;qRT-PCR results showed that bevacizumab inhibited mRNA expression of MDR1,MRP1 and P-gp in HNE1/NDP cells after treatment.PI staining showed that bevacizumab promoted apoptosis in HNE1/NDP cells.Conclusions Bevacizuma b reverses the drug resistance via down-regulates the expression of multidrug resistance related genes and promotes apoptosis of HNE1/NDP.

关 键 词：鼻咽癌 贝伐单抗 耐药逆转 细胞凋亡 

分 类 号：R739.62[医药卫生—肿瘤]