中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2014年
1期
5-10
,共6页
赵潇%任贺%高松%郝继辉
趙瀟%任賀%高鬆%郝繼輝
조소%임하%고송%학계휘
胰腺肿瘤%薯蓣皂苷%细胞凋亡%过氧化还原酶1
胰腺腫瘤%藷蕷皂苷%細胞凋亡%過氧化還原酶1
이선종류%서여조감%세포조망%과양화환원매1
Pancreatic neoplasms%Dioscin%Apoptosis%Peroxiredoxin-1
目的 探讨薯蓣皂苷对胰腺癌细胞MiaPaCa-2凋亡的作用及其对细胞中过氧化还原酶1(PRDX1)表达水平的影响.方法 采用四甲基偶氮唑蓝法检测不同浓度薯蓣皂苷对MiaPaCa-2细胞增殖抑制的影响,流式细胞术检测细胞的凋亡,Western blot检测PRDX1以及凋亡相关蛋白的表达,DCFH-DA荧光探针检测细胞中活性氧簇(ROS)的水平.结果 薯蓣皂苷可以抑制MiaPaCa-2细胞的增殖,且有剂量-效应依赖关系.薯蓣皂苷可以诱导MiaPaCa-2细胞内ROS水平明显上升.2.5 μmol/L和5μmol/L薯蓣皂苷处理MiaPaCa-2细胞12 h后,细胞凋亡率分别为(28.4±0.9)％和(49.6±2.7)％,与空白对照组[(3.5±0.7)％]比较,差异均有统计学意义(均P ＜0.05).Western blot检测结果显示,与对照组比较,薯蓣皂苷组的抗凋亡蛋白表达降低,而凋亡蛋白表达上调,但加入N-乙酰半胱氨酸(NAC)后,薯蓣皂苷组的这种诱导作用受到抑制.10 mmol/L NAC和2.5 μ.mol/L薯蓣皂苷、10 mmol/L NAC和5μmol/L薯蓣皂苷处理MiaPaCa-2细胞12h后,凋亡率分别为(10.8±2.3)％和(18.8 ±3.0)％.薯蓣皂苷可以降低MiaPaCa-2细胞的PRDX1蛋白表达.过表达PRDXI后,薯蓣皂苷诱导MiaPaCa-2细胞内ROS水平上升和凋亡的作用明显受到抑制,凋亡率为(21.3±5.9)％.结论 薯蓣皂苷可以下调胰腺癌MiaPaCa-2细胞中PRDX1的表达水平,进而导致ROS过度蓄积引起细胞凋亡.
目的 探討藷蕷皂苷對胰腺癌細胞MiaPaCa-2凋亡的作用及其對細胞中過氧化還原酶1(PRDX1)錶達水平的影響.方法 採用四甲基偶氮唑藍法檢測不同濃度藷蕷皂苷對MiaPaCa-2細胞增殖抑製的影響,流式細胞術檢測細胞的凋亡,Western blot檢測PRDX1以及凋亡相關蛋白的錶達,DCFH-DA熒光探針檢測細胞中活性氧簇(ROS)的水平.結果 藷蕷皂苷可以抑製MiaPaCa-2細胞的增殖,且有劑量-效應依賴關繫.藷蕷皂苷可以誘導MiaPaCa-2細胞內ROS水平明顯上升.2.5 μmol/L和5μmol/L藷蕷皂苷處理MiaPaCa-2細胞12 h後,細胞凋亡率分彆為(28.4±0.9)％和(49.6±2.7)％,與空白對照組[(3.5±0.7)％]比較,差異均有統計學意義(均P ＜0.05).Western blot檢測結果顯示,與對照組比較,藷蕷皂苷組的抗凋亡蛋白錶達降低,而凋亡蛋白錶達上調,但加入N-乙酰半胱氨痠(NAC)後,藷蕷皂苷組的這種誘導作用受到抑製.10 mmol/L NAC和2.5 μ.mol/L藷蕷皂苷、10 mmol/L NAC和5μmol/L藷蕷皂苷處理MiaPaCa-2細胞12h後,凋亡率分彆為(10.8±2.3)％和(18.8 ±3.0)％.藷蕷皂苷可以降低MiaPaCa-2細胞的PRDX1蛋白錶達.過錶達PRDXI後,藷蕷皂苷誘導MiaPaCa-2細胞內ROS水平上升和凋亡的作用明顯受到抑製,凋亡率為(21.3±5.9)％.結論 藷蕷皂苷可以下調胰腺癌MiaPaCa-2細胞中PRDX1的錶達水平,進而導緻ROS過度蓄積引起細胞凋亡.
목적 탐토서여조감대이선암세포MiaPaCa-2조망적작용급기대세포중과양화환원매1(PRDX1)표체수평적영향.방법 채용사갑기우담서람법검측불동농도서여조감대MiaPaCa-2세포증식억제적영향,류식세포술검측세포적조망,Western blot검측PRDX1이급조망상관단백적표체,DCFH-DA형광탐침검측세포중활성양족(ROS)적수평.결과 서여조감가이억제MiaPaCa-2세포적증식,차유제량-효응의뢰관계.서여조감가이유도MiaPaCa-2세포내ROS수평명현상승.2.5 μmol/L화5μmol/L서여조감처리MiaPaCa-2세포12 h후,세포조망솔분별위(28.4±0.9)％화(49.6±2.7)％,여공백대조조[(3.5±0.7)％]비교,차이균유통계학의의(균P ＜0.05).Western blot검측결과현시,여대조조비교,서여조감조적항조망단백표체강저,이조망단백표체상조,단가입N-을선반광안산(NAC)후,서여조감조적저충유도작용수도억제.10 mmol/L NAC화2.5 μ.mol/L서여조감、10 mmol/L NAC화5μmol/L서여조감처리MiaPaCa-2세포12h후,조망솔분별위(10.8±2.3)％화(18.8 ±3.0)％.서여조감가이강저MiaPaCa-2세포적PRDX1단백표체.과표체PRDXI후,서여조감유도MiaPaCa-2세포내ROS수평상승화조망적작용명현수도억제,조망솔위(21.3±5.9)％.결론 서여조감가이하조이선암MiaPaCa-2세포중PRDX1적표체수평,진이도치ROS과도축적인기세포조망.
Objective The aim of this study was to observe the effects of dioscin on apoptosis and on expression of PRDX1 in pancreatic cancer MiaPaCa-2 cells in vitro.Methods MTT assay was used to detect the growth rate among the medication groups treated with different concentrations of dioscin.The apoptosis rate was determined by annexin V-fluorescein isothiocyanate/propidium iodide double staining and flow cytometry.Western blot analysis was used to assay the expression of PRDX1 and apoptotic proteins in the cells.Reactive oxygen species (ROS) formation was measured by 2'7'-dichlorofluorescein diacetate (DCFH-DA).Results Dioscin considerably inhibited the proliferation of MiaPaCa-2 cells in vitro.The inhibitory action was enhanced in a dose-dependent manner.The levels of intracellular ROS detected with DCFH-DA were highly increased after dioscin treatment.The flow cytometry analysis using annexiu V-PI staining showed that compared with the apoptotic rate of control group [(3.5 ± 0.7) ％],2.5 μmol/L and 5 μmol/L dioscin induced apoptosis in (28.4 ± 0.9) ％ and (49.6 ± 2.7) ％ MiaPaCa-2 cells,and Western blot analysis showed that apoptotic proteins Bax and cleaved caspase-3 expressions were increased and antiapoptotic protein Bcl-2 expression was decreased.In addition,these effects could be blocked by antioxidant N-acetylcysteine (NAC) administration,and the apoptotic rates decreased to (10.8 ± 2.3)％ and (18.8 ± 3.0) ％,respectively.We further observed the decrease of PRDX1 expression after dioscin treatment.Moreover,after PRDX1 overexpression,dioscin treatment no longer induced high levels of ROS and apoptosis,and the apoptotic rate was decreased to (21.3 ± 5.9) ％.Conclusion Dioscin can downregulate the PRDX1 expression,and then induces ROS-mediated apoptosis in cancer cells.
