PPAR-γ对人鼻咽癌 CNE1／DDP 细胞顺铂耐药性的逆转作用及其机制探讨

摘要

Objective To investigate the effect of PPAR-γon cisplatin sensitivity reversal of human nasopharyngeal carcinoma CNE1/DDP cell line and its mechanism.Methods The human nasopharyngeal carcinoma CNE1/DDP cells were treated by 10 μg/mL and 20 μg/mL PPAR-γagonist rosiglitazone.MTS assay was used to measure the effect of pro-liferation;flow cytometry was used to measure the cell apoptosis and P-gp expression;RT-PCR and Western blot assay were used to measure the mRNA and protein expression of multidrug resistance gene (MDR1), multidrug resistance-associated protein ( MRP) , B-cell lymphoma gene 2 ( bcl-2);Western blot assay was used to measure the phosphorylation of Akt ( p-Akt).Results CNE1/DDP cells were treated by 10μg/mL and 20μg/mL rosiglitazone, the drug resistance reverse rates were 100.0%, 149.3%±11.3%, 293.8%±25.5%, respectively;the apoptosis rate was gradually increased;the mR-NA and protein expression of PPAR-γ, MDR1, MRP and bcl-2, and p-Akt expression were gradually decreased (all P<0.05).Conclusions PPAR-γcould increase cisplatin sensitivity of CNE1/DDP cells by down-regulating phosphorylation of Akt and the expression of MDR1 and MRP.%目的：观察氧化物酶体增殖物激活受体-γ（ PPAR-γ）对人鼻咽癌CNE1/DDP细胞顺铂耐药性的逆转作用，并探讨其机制。方法将CNE1/DDP细胞分别加入10、20μg/mL PPAR-γ激动剂罗格列酮和等体积培养液进行培养。采用MTS法检测细胞耐药逆转率，流式细胞术检测细胞凋亡率和P-糖蛋白（ P-gp）表达，RT-PCR法和Western blot法检测多重耐药基因（MDR1）、多药耐药相关蛋白基因（MRP）和B淋巴细胞瘤-2基因（bcl-2）mRNA和蛋白表达，Western blot法检测磷酸化Akt蛋白（ p-Akt ）。结果加入等体积培养液和10、20μg/mL罗格列酮的CNE1/DDP细胞耐药逆转率分别为100．0％、149．3％±11．3％、293．8％±25．5％，其细胞凋亡率依次升高，PPAR-γ、MDR1、MRP、bcl-2 mRNA和蛋白表达以及p-gp阳性率、p-Akt表达依次降低；两两比较，P均＜0．05。结论 PPAR-γ对CNE1/DDP细胞的顺铂耐药性具有逆转作用，且呈剂量依赖性；可能与其抑制Akt磷酸化和耐药基因MDR1、MRP表达，并促进细胞凋亡有关。