摘要
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。
Objective To investigate the effect of PPAR-γ on 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-γ agonist rosiglitazone. MTS assay was used to measure the effect of proliferation; 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（pAkt）. Results CNE1 /DDP cells were treated by 10 μg /mL and 20 μg /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 mRNA and protein expression of PPAR-γ,MDR1,MRP and bcl-2,and p-Akt expression were gradually decreased（all P〈0. 05）. Conclusions PPAR-γ could increase cisplatin sensitivity of CNE1 /DDP cells by down-regulating phosphorylation of Akt and the expression of MDR1 and MRP.
出处
《山东医药》
CAS
2014年第40期10-12,15,共4页
Shandong Medical Journal
