中国肿瘤临床
中國腫瘤臨床
중국종류림상
CHINESE JOURNAL OF CLINICAL ONCOLOGY
2009年
21期
1243-1247
,共5页
张燕%祝淑钗%赵蔚然%董英辉%张献波%乞国艳
張燕%祝淑釵%趙蔚然%董英輝%張獻波%乞國豔
장연%축숙차%조위연%동영휘%장헌파%걸국염
褪黑素%乳腺癌%阿霉素%增敏作用%p53%bcl-2
褪黑素%乳腺癌%阿黴素%增敏作用%p53%bcl-2
퇴흑소%유선암%아매소%증민작용%p53%bcl-2
Melatonin%Breast cancer%Adriamycin%Sensitization%p53%bcl-2
目的:探讨生理浓度(10~(-9)mol/L)褪黑素和药理浓度(10~(-5)mol/L)褪黑素对阿霉素抑制ER~+乳腺癌细胞MCF-7作用影响及其机制.方法:1)应用四甲基偶氮唑蓝(MTT)法检测经不同浓度褪黑素孵育后阿霉素对MCF-7的抑制率和IC_(50)变化.2)应用流式细胞学方法观察不同浓度褪黑素、阿霉素以及两药联用时对MCF-7凋亡的影响.3)应用Western blotting法检测不同浓度褪黑素、阿霉素单药和两药联用对MCF-7细胞p53和bcl-2蛋白表达的影响.结果:1)阿霉素对MCF-7乳腺癌细胞具有明显的抑制作用,且成剂量时间依赖性.IC_(50)值为0.62±0.07μg/ml,经生理浓度和药理浓度褪黑素孵育后IC_(50)值分别降为0.59±0.09μg/ml和0.42±0.02μg/ml,前者与孵育前相比未见统计学差异(P>0.05),后者相比差异显著(P<0.01).2)流式细胞学检测结果显示:阿霉素对MCF-7细胞具有凋亡促进作用,随浓度增高凋亡率未见增加(P>0.05).生理浓度褪黑素联合阿霉素与相应浓度的阿霉素单药相比,凋亡率未见明显增加(P>0.05).药理浓度褪黑素联合阿霉素与相应浓度阿霉素单药比较,凋亡率明显增加(P<0.05),联合组随着阿霉素浓度增加凋亡率并未见明显变化(P>0.05).3)MCF-7乳腺癌细胞株中呈p53蛋白低表达、bcl-2蛋白高表达.生理浓度褪黑素即可显著增高MCF-7细胞p53蛋白并降低bcl-2蛋白表达,并有剂量依赖性(P<0.01).药理浓度褪黑素联合不同浓度阿霉素对两个蛋白表达未见显著性差异(P>0.05);阿霉素对两种蛋白表达未见明显影响(P>0.05).结论:1)生理浓度褪黑素(10~(-9)mol/L)对阿霉素的抗癌作用未见明显影响,药理浓度(10~(-5)mol/L)以上褪黑素表现出对阿霉素明显的增敏作用.2)阿霉素较低浓度时,凋亡促进作用可能是褪黑素对其增敏机制的一部分,随着阿霉素浓度的提高,褪黑素的细胞毒增敏机制可能占主要地位.3)生理浓度褪黑素能提高ER~+乳腺癌细胞p53蛋白表达并降低bcl-2蛋白表达,并具有剂量依赖性.涉及p53和bcl-2的凋亡通路可能是褪黑素对阿霉素增敏机制的一部分.
目的:探討生理濃度(10~(-9)mol/L)褪黑素和藥理濃度(10~(-5)mol/L)褪黑素對阿黴素抑製ER~+乳腺癌細胞MCF-7作用影響及其機製.方法:1)應用四甲基偶氮唑藍(MTT)法檢測經不同濃度褪黑素孵育後阿黴素對MCF-7的抑製率和IC_(50)變化.2)應用流式細胞學方法觀察不同濃度褪黑素、阿黴素以及兩藥聯用時對MCF-7凋亡的影響.3)應用Western blotting法檢測不同濃度褪黑素、阿黴素單藥和兩藥聯用對MCF-7細胞p53和bcl-2蛋白錶達的影響.結果:1)阿黴素對MCF-7乳腺癌細胞具有明顯的抑製作用,且成劑量時間依賴性.IC_(50)值為0.62±0.07μg/ml,經生理濃度和藥理濃度褪黑素孵育後IC_(50)值分彆降為0.59±0.09μg/ml和0.42±0.02μg/ml,前者與孵育前相比未見統計學差異(P>0.05),後者相比差異顯著(P<0.01).2)流式細胞學檢測結果顯示:阿黴素對MCF-7細胞具有凋亡促進作用,隨濃度增高凋亡率未見增加(P>0.05).生理濃度褪黑素聯閤阿黴素與相應濃度的阿黴素單藥相比,凋亡率未見明顯增加(P>0.05).藥理濃度褪黑素聯閤阿黴素與相應濃度阿黴素單藥比較,凋亡率明顯增加(P<0.05),聯閤組隨著阿黴素濃度增加凋亡率併未見明顯變化(P>0.05).3)MCF-7乳腺癌細胞株中呈p53蛋白低錶達、bcl-2蛋白高錶達.生理濃度褪黑素即可顯著增高MCF-7細胞p53蛋白併降低bcl-2蛋白錶達,併有劑量依賴性(P<0.01).藥理濃度褪黑素聯閤不同濃度阿黴素對兩箇蛋白錶達未見顯著性差異(P>0.05);阿黴素對兩種蛋白錶達未見明顯影響(P>0.05).結論:1)生理濃度褪黑素(10~(-9)mol/L)對阿黴素的抗癌作用未見明顯影響,藥理濃度(10~(-5)mol/L)以上褪黑素錶現齣對阿黴素明顯的增敏作用.2)阿黴素較低濃度時,凋亡促進作用可能是褪黑素對其增敏機製的一部分,隨著阿黴素濃度的提高,褪黑素的細胞毒增敏機製可能佔主要地位.3)生理濃度褪黑素能提高ER~+乳腺癌細胞p53蛋白錶達併降低bcl-2蛋白錶達,併具有劑量依賴性.涉及p53和bcl-2的凋亡通路可能是褪黑素對阿黴素增敏機製的一部分.
목적:탐토생리농도(10~(-9)mol/L)퇴흑소화약리농도(10~(-5)mol/L)퇴흑소대아매소억제ER~+유선암세포MCF-7작용영향급기궤제.방법:1)응용사갑기우담서람(MTT)법검측경불동농도퇴흑소부육후아매소대MCF-7적억제솔화IC_(50)변화.2)응용류식세포학방법관찰불동농도퇴흑소、아매소이급량약련용시대MCF-7조망적영향.3)응용Western blotting법검측불동농도퇴흑소、아매소단약화량약련용대MCF-7세포p53화bcl-2단백표체적영향.결과:1)아매소대MCF-7유선암세포구유명현적억제작용,차성제량시간의뢰성.IC_(50)치위0.62±0.07μg/ml,경생리농도화약리농도퇴흑소부육후IC_(50)치분별강위0.59±0.09μg/ml화0.42±0.02μg/ml,전자여부육전상비미견통계학차이(P>0.05),후자상비차이현저(P<0.01).2)류식세포학검측결과현시:아매소대MCF-7세포구유조망촉진작용,수농도증고조망솔미견증가(P>0.05).생리농도퇴흑소연합아매소여상응농도적아매소단약상비,조망솔미견명현증가(P>0.05).약리농도퇴흑소연합아매소여상응농도아매소단약비교,조망솔명현증가(P<0.05),연합조수착아매소농도증가조망솔병미견명현변화(P>0.05).3)MCF-7유선암세포주중정p53단백저표체、bcl-2단백고표체.생리농도퇴흑소즉가현저증고MCF-7세포p53단백병강저bcl-2단백표체,병유제량의뢰성(P<0.01).약리농도퇴흑소연합불동농도아매소대량개단백표체미견현저성차이(P>0.05);아매소대량충단백표체미견명현영향(P>0.05).결론:1)생리농도퇴흑소(10~(-9)mol/L)대아매소적항암작용미견명현영향,약리농도(10~(-5)mol/L)이상퇴흑소표현출대아매소명현적증민작용.2)아매소교저농도시,조망촉진작용가능시퇴흑소대기증민궤제적일부분,수착아매소농도적제고,퇴흑소적세포독증민궤제가능점주요지위.3)생리농도퇴흑소능제고ER~+유선암세포p53단백표체병강저bcl-2단백표체,병구유제량의뢰성.섭급p53화bcl-2적조망통로가능시퇴흑소대아매소증민궤제적일부분.
Objective: To investigate the sensitization of physiological (10~(-9)mol/L) and pharmacological (10~(-5)mol/L) concentrations of melatonin on cell line MCF-7 for adriamycin and its mechanism. Methods: (1) MTT was applied to test the changes in inhibition ratio and IC_(50) of call line MCF-7 for adriamycin before and af-ter incubation with melatonin. (2) Flow cytometry was used to observe the effect of different concentrations of melatonin, adriamycin and melatonin plus adriamycin on cell apoptosis. (3) Western blot was employed to de-termine the expression of p53 and bcl-2 in MCF-7 cells incubated with melatonin, adriamycin and melatonin plus adriamycin. Results: (1) MTT method showed that adriamycin had inhibitive effect on the growth of MCF-7 cells in a dose- and time-dependent manner. The IC_(50) of cell line MCF-7 for adnamycin before treat-ment with melatonin was 0.62±0.07ug/mL (P>0.05). The IC50 of cell line MCF-7 for adriamycin incubated with physiological and pharmacological concentrations of melatonin was 0.59±0.09ug/mL and 0.42±0.02ug/mL, re-spectively, with a significant difference (P<0.01). (2) Flow cytometry method showed that adriamycin could promote apoptosis of MCF-7, and no changes in the apoptosis index were observed as the concentration of melatonin was changed (P>0.05). With the same concentration of adriamycin, the apoptosis index of cells treated with physiological concentration of melatonin plus adriamycin was not changed (P=>0.05), but the apop-tosis index of cells treated with pharmacological concentrations of melatonin plus addamycin was increased significantly. The concentration of adriamycin had no effect on the apoptosis index. (3) Western blot showed that P53 protein was expressed at a lower level and bcl-2 protein was highly expressed. Physiological concen-trations of melatonin increased the expression of p53 and decreased bcl-2 expression in a dose - dependent manner. The concentration of addamycin had no effect on the expression of p53 and bcl-2 proteins. Conclu-sion: (1) Physiological concentrations of melatonin had no effect on the anti-cancer effect of adriamycin. Phar-macological concentrations of melatonin showed sensitization of MCF-7 cells for adriamycin. (2) With a lower concentration of adriamycin, the promotion of apoptosis may be part of the mechanism of sensitization effect of melatonin. With the increase of adriamycin concentration, the cytotoxic mechanism of melotonin became more and more important. (3) Physiological concentration could increase the expression of p53 and decrease bcl-2 expression in ER~+ breast carcinoma cell line MCF-7 in a dose -dependent manner. The apoptosis involv-ing p53 and bcl-2 passway was part of the mechanism of sensitization effect of melatonin for addamycin.
