安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
ACTA UNIVERSITY MEDICINALIS ANHUI
2014年
10期
1404-1408
,共5页
依西美坦%甲氨蝶呤%耐药细胞%乳腺癌
依西美坦%甲氨蝶呤%耐藥細胞%乳腺癌
의서미탄%갑안접령%내약세포%유선암
exemestane%methotrexate%drug-resistance cell%breast cancer
目的研究依西美坦( EXE )与低剂量甲氨蝶呤( MTX)对依西美坦耐药人乳腺癌 MCF-7细胞( MCF-7/EXE)的增殖抑制作用及意义。方法使用 MTT 法检测EXE与低剂量MTX单药和联合给药对MCF-7/EXE细胞的增殖抑制作用,计算依西美坦与低剂量甲氨蝶呤联合给药时的联合指数(CI),评价两药间的相互作用。流式细胞术检测IC50浓度药物EXE组(120μmol/L)、低剂量MTX组(60 nmol/L)、EXE+MTX组处理后的MCF-7/EXE细胞周期变化。荧光染色观察单药及联合作用对细胞凋亡的影响。Western blot法检测各组细胞 Bcl-2、AKT、P-AKT、COX-2蛋白的表达情况。结果 MTT法表明,与单药作用相比,联合治疗明显降低 MCF-7/EXE细胞的活性( CI<0.9)。此外, EXE和低剂量MTX的联合使用表现对细胞增殖的协同抑制作用,细胞周期显著阻滞于S期,Western blot法显示联合组与对照组或单个药物组相比对P-ATK、Bcl-2、COX-2蛋白的表达有更强的抑制作用。结论 EXE与低剂量MTX联合使用时对MCF-7/EXE细胞产生协同抑制作用,可逆转耐药。
目的研究依西美坦( EXE )與低劑量甲氨蝶呤( MTX)對依西美坦耐藥人乳腺癌 MCF-7細胞( MCF-7/EXE)的增殖抑製作用及意義。方法使用 MTT 法檢測EXE與低劑量MTX單藥和聯閤給藥對MCF-7/EXE細胞的增殖抑製作用,計算依西美坦與低劑量甲氨蝶呤聯閤給藥時的聯閤指數(CI),評價兩藥間的相互作用。流式細胞術檢測IC50濃度藥物EXE組(120μmol/L)、低劑量MTX組(60 nmol/L)、EXE+MTX組處理後的MCF-7/EXE細胞週期變化。熒光染色觀察單藥及聯閤作用對細胞凋亡的影響。Western blot法檢測各組細胞 Bcl-2、AKT、P-AKT、COX-2蛋白的錶達情況。結果 MTT法錶明,與單藥作用相比,聯閤治療明顯降低 MCF-7/EXE細胞的活性( CI<0.9)。此外, EXE和低劑量MTX的聯閤使用錶現對細胞增殖的協同抑製作用,細胞週期顯著阻滯于S期,Western blot法顯示聯閤組與對照組或單箇藥物組相比對P-ATK、Bcl-2、COX-2蛋白的錶達有更彊的抑製作用。結論 EXE與低劑量MTX聯閤使用時對MCF-7/EXE細胞產生協同抑製作用,可逆轉耐藥。
목적연구의서미탄( EXE )여저제량갑안접령( MTX)대의서미탄내약인유선암 MCF-7세포( MCF-7/EXE)적증식억제작용급의의。방법사용 MTT 법검측EXE여저제량MTX단약화연합급약대MCF-7/EXE세포적증식억제작용,계산의서미탄여저제량갑안접령연합급약시적연합지수(CI),평개량약간적상호작용。류식세포술검측IC50농도약물EXE조(120μmol/L)、저제량MTX조(60 nmol/L)、EXE+MTX조처리후적MCF-7/EXE세포주기변화。형광염색관찰단약급연합작용대세포조망적영향。Western blot법검측각조세포 Bcl-2、AKT、P-AKT、COX-2단백적표체정황。결과 MTT법표명,여단약작용상비,연합치료명현강저 MCF-7/EXE세포적활성( CI<0.9)。차외, EXE화저제량MTX적연합사용표현대세포증식적협동억제작용,세포주기현저조체우S기,Western blot법현시연합조여대조조혹단개약물조상비대P-ATK、Bcl-2、COX-2단백적표체유경강적억제작용。결론 EXE여저제량MTX연합사용시대MCF-7/EXE세포산생협동억제작용,가역전내약。
Objective To investigate the combined effect of exemestane and low-dose methotrexate on exemestane-resistant MCF-7 human breast cancer cells( MCF-7/EXE). Methods Antiproliferative effects of exemestane and low-dose of methotrexate, alone and in combination on growth of MCF-7/EXE cells were assessed by using the MTT assay. Synergistic interaction between the two drugs was evaluated in vitro by using the combination index ( CI) method. The cell cycle distribution was analyzed by flow cytometry in a half inhibitory concentration of exemestane and low-dose of methotrexate . The changes of apoptosis on MCF-7/EXE cells exposed to two drugs alone or in com-bination were observed by fluorescence microscope. The expression of Bcl-2,AKT,P-AKT and cyclooxygenase-2 was investigated by Western blot. Results MTT assays indicated that the combination treatment apparently decreased the viability of MCF-7/EXE cells compared to single drug treatment (CI<0. 9). In addition, the combination of exemestane and low-dose methotrexate exhibited a synergistic inhibition of cell proliferation, arrested the cell cycle in the S phase significantly and produced a stronger inhibitory effects on P-AKT, Bcl-2 and cyclooxygenase-2 ex-pression than control or individual drug treatment. Conclusion The combination of the two inhibitors significantly increases the response as compared to single agent treatment, suggesting that combination treatment which can re-verse the resistance of exemestane could be a more effective approach to breast cancer.
