CAJ | 학술논문

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(99m technetium-methoxyisobutyli-sonitrile,99m Tc-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99m Tc-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99m Tc-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99m Tc-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99m Tc-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。
목적：탐토2-탈양-D-포도당(2-DG)대비인암순박내약주세포( HNE-1/DDP )다약내약( MDR )적역전궤제,관찰99m득-갑양기이정기이정(99m technetium-methoxyisobutyli-sonitrile,99m Tc-MIBI)세포내적섭취변화,평개2-DG역전종류세포다약내약적효과。방법γ계수기검측불동농도2-DG작용하HNE-1/DDP세포대99m Tc-MIBI적섭취청제정황이급2-DG농도위10 mmol·L-1시HNE-1급HNE-1/DDP세포대99m Tc-MIBI적섭취청제정황。측정2-DG 작용하HNE-1/DDP세포내ATP치。 Western blot검측2-DG작용하HNE-1/DDP세포내P당단백( P-gp)、다약내약상관단백( MRP)표체。추화병정( PI)단염검측순박( DDP)단독급연합2-DG작용하HNE-1/DDP세포적조망정황。결과 HNE-1/DDP세포대99m Tc-MIBI적청제솔위54．8%,고우HNE-1세포적청제솔-41．3%(P <0．01),2-DG(10 mmol·L-1)작용후HNE-1/DDP세포적청제솔위-203．7%,교전명현강저( P<0．01)。 HNE-1/DDP세포재2-DG작용하 ATP치위대조조적55．69%,차P-gp、MRP단백표체교대조조감소。 DDP연합2-DG(10 mmol·L-1)작용24 h후HNE-1/DDP세포조망솔위49．4%,고우단독사용DDP시HNE-1/DDP세포적조망솔(22．5%)。결론99m Tc-MIBI가유효검측HNE-1/DDP세포다약내약현상급평개2-DG적역전효과,2-DG역전궤제가능여세포내ATP생성억제급상관전운단백표체감소유관。
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.