摘要
目的探讨2-脱氧-D-葡萄糖(2-DG)对鼻咽癌顺铂耐药株细胞(HNE-1/DDP)多药耐药(MDR)的逆转机制,观察99m锝-甲氧基异丁基异腈(99mtechnetium-methoxyisobutylisonitrile,99mTc-MIBI)细胞内的摄取变化,评价2-DG逆转肿瘤细胞多药耐药的效果。方法γ计数器检测不同浓度2-DG作用下HNE-1/DDP细胞对99mTc-MIBI的摄取清除情况以及2-DG浓度为10 mmol·L-1时HNE-1及HNE-1/DDP细胞对99mTc-MIBI的摄取清除情况。测定2-DG作用下HNE-1/DDP细胞内ATP值。Western blot检测2-DG作用下HNE-1/DDP细胞内P糖蛋白(P-gp)、多药耐药相关蛋白(MRP)表达。溴化丙啶(PI)单染检测顺铂(DDP)单独及联合2-DG作用下HNE-1/DDP细胞的凋亡情况。结果 HNE-1/DDP细胞对99mTc-MIBI的清除率为54.8%,高于HNE-1细胞的清除率-41.3%(P<0.01),2-DG(10 mmol·L-1)作用后HNE-1/DDP细胞的清除率为-203.7%,较前明显降低(P<0.01)。HNE-1/DDP细胞在2-DG作用下ATP值为对照组的55.69%,且P-gp、MRP蛋白表达较对照组减少。DDP联合2-DG(10 mmol·L-1)作用24 h后HNE-1/DDP细胞凋亡率为49.4%,高于单独使用DDP时HNE-1/DDP细胞的凋亡率(22.5%)。结论99mTc-MIBI可有效检测HNE-1/DDP细胞多药耐药现象及评价2-DG的逆转效果,2-DG逆转机制可能与细胞内ATP生成抑制及相关转运蛋白表达减少有关。
Aim To evaluate the reversal effect of 2-deoxy-D-glucose ( 2-DG ) on multidrug resistance ( MDR) by observing the uptake change of 99m Tc-MIBI in HNE-1/DDP cells, and to explore its mechanism. Methods The uptake of 99m Tc-MIBI in HNE-1/DDP cells under different concentrations of 2-DG was detec-ted by γ-counter, and the clearance rates of 99m Tc-MI-BI in HNE-1 cells and HNE-1/DDP cells after treated with 2-DG (10 mmol·L-1 ) were compared. The con-tent of ATP in HNE-1/DDP cells was detected after treated with 2-DG. P-glycoprotein ( P-gp ) and multi-drug resistance-associated proteins ( MRP ) expression were measured by Western blot. Apoptotic HNE-1/DDP cells treated with DDP alone or combined with 2-DG (10 mmol·L-1 ) were detected by propidium io-dide ( PI ) staining. Results The clearance rate of 99m Tc-MIBI in HNE-1/DDP cells was 54. 8%, which was significantly higher than that ( - 41. 3%) in HNE-1 cells (P<0. 01). The clearance rate of 99mTc-MIBI in HNE-1/DDP cells was -203. 7% after treat-ment with 2-DG ( 10 mmol · L-1 ) , which could be significantly reduced compared with the control group ( P<0. 01 ) . The level of ATP was 55 . 69% compared with the negative control group and the expression of P-gp and MRP protein decreased dramatically in HNE-1/DDP. With the combination of 2-DG and DDP, the ap-optotic rate of HNE-1/DDP cells reached 49 . 4%which was significantly higher than DDP treated group (22. 5%) . Conclusion Multidrug resistance and the reversal effect of 2-DG on multidrug resistance could be evaluated effectively by detecting the uptake change of 99m Tc-MIBI in HNE-1/DDP cells. The mechanism may be related with the inhibition of ATP level and the re-duced expression of P-gp and MRP protein in cancer cells.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2015年第10期1433-1438,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81371587)
安徽省高等学校自然科学研究项目(No KJ2015B071by)
