中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2014年
2期
106-109
,共4页
吴小进%米燕燕%杨辉%冯青青%董海北%殷咏梅
吳小進%米燕燕%楊輝%馮青青%董海北%慇詠梅
오소진%미연연%양휘%풍청청%동해북%은영매
黄芪多糖%乳腺肿瘤%细胞增殖%细胞凋亡
黃芪多糖%乳腺腫瘤%細胞增殖%細胞凋亡
황기다당%유선종류%세포증식%세포조망
Astragalan%Breast neoplasms%Cell proliferation%Apoptosis
目的 探讨中药黄芪多糖的体外抗人乳腺癌MCF-7细胞活性.方法 实验分为空白对照组、黄芪多糖组和阳性对照组,黄芪多糖组MCF-7细胞给予不同浓度(2.5、5、10、20 mg/L)的黄芪多糖,阳性对照组给予10 μmol/L顺铂,空白对照组给予等体积培养基.48 h后应用四甲基偶氮唑蓝(MTT)法测黄芪多糖对MCF-7细胞增殖抑制率,计算IC50;吖啶橙(AO)/溴化乙啶(EB)荧光染色法测定黄芪多糖对MCF-7细胞诱导凋亡作用;应用流式细胞仪分析黄芪多糖对MCF-7细胞凋亡和细胞周期的影响.结果 在给予2.5、5、10、20 mg/L黄芪多糖48 h后,黄芪多糖呈浓度依赖性抑制MCF-7细胞的增殖(r=0.985,P＜0.05),抑制率分别为(4.14±2.96)％、(7.14±2.10)％、(20.13±2.33)％、(64.66±5.15)％,高于空白对照组0％,但4个不同浓度组的抑制率均低于阳性对照组(90.31±4.92)％.黄芪多糖48 h的IC50=16.83m/L.随着黄芪多糖浓度的逐渐增高,MCF-7细胞中代表凋亡的玛瑙色逐渐增多,细胞核骤缩、核分裂,细胞形态呈现典型的凋亡特征.2.5、5、10、20 mg/L黄芪多糖的凋亡率分别为(2.37±0.98)％、(6.76±1.31)％、(11.65±1.46)％、(20.75±2.68)％,高于空白对照组(1.14±1.25)％(均P＜0.05),但均低于阳性对照组(35.09±2.88)％(均P＜ 0.05).与空白对照组比较,黄芪多糖以浓度依赖性诱导MCF-7细胞凋亡(r=0.991,P＜0.05),随浓度的增加,细胞凋亡率升高,但均低于阳性对照组.黄芪多糖随浓度的增加促使S期细胞比例逐渐升高,但低于空白和阳性对照组(均P＜0.05),使处于G0-G1期细胞的比例逐渐减少,仍高于空白和阳性对照组(均P＜0.05).结论 黄芪多糖抑制人乳腺癌MCF-7细胞增殖并诱导其凋亡,使MCF-7细胞生长增殖停滞在S期.
目的 探討中藥黃芪多糖的體外抗人乳腺癌MCF-7細胞活性.方法 實驗分為空白對照組、黃芪多糖組和暘性對照組,黃芪多糖組MCF-7細胞給予不同濃度(2.5、5、10、20 mg/L)的黃芪多糖,暘性對照組給予10 μmol/L順鉑,空白對照組給予等體積培養基.48 h後應用四甲基偶氮唑藍(MTT)法測黃芪多糖對MCF-7細胞增殖抑製率,計算IC50;吖啶橙(AO)/溴化乙啶(EB)熒光染色法測定黃芪多糖對MCF-7細胞誘導凋亡作用;應用流式細胞儀分析黃芪多糖對MCF-7細胞凋亡和細胞週期的影響.結果 在給予2.5、5、10、20 mg/L黃芪多糖48 h後,黃芪多糖呈濃度依賴性抑製MCF-7細胞的增殖(r=0.985,P＜0.05),抑製率分彆為(4.14±2.96)％、(7.14±2.10)％、(20.13±2.33)％、(64.66±5.15)％,高于空白對照組0％,但4箇不同濃度組的抑製率均低于暘性對照組(90.31±4.92)％.黃芪多糖48 h的IC50=16.83m/L.隨著黃芪多糖濃度的逐漸增高,MCF-7細胞中代錶凋亡的瑪瑙色逐漸增多,細胞覈驟縮、覈分裂,細胞形態呈現典型的凋亡特徵.2.5、5、10、20 mg/L黃芪多糖的凋亡率分彆為(2.37±0.98)％、(6.76±1.31)％、(11.65±1.46)％、(20.75±2.68)％,高于空白對照組(1.14±1.25)％(均P＜0.05),但均低于暘性對照組(35.09±2.88)％(均P＜ 0.05).與空白對照組比較,黃芪多糖以濃度依賴性誘導MCF-7細胞凋亡(r=0.991,P＜0.05),隨濃度的增加,細胞凋亡率升高,但均低于暘性對照組.黃芪多糖隨濃度的增加促使S期細胞比例逐漸升高,但低于空白和暘性對照組(均P＜0.05),使處于G0-G1期細胞的比例逐漸減少,仍高于空白和暘性對照組(均P＜0.05).結論 黃芪多糖抑製人乳腺癌MCF-7細胞增殖併誘導其凋亡,使MCF-7細胞生長增殖停滯在S期.
목적 탐토중약황기다당적체외항인유선암MCF-7세포활성.방법 실험분위공백대조조、황기다당조화양성대조조,황기다당조MCF-7세포급여불동농도(2.5、5、10、20 mg/L)적황기다당,양성대조조급여10 μmol/L순박,공백대조조급여등체적배양기.48 h후응용사갑기우담서람(MTT)법측황기다당대MCF-7세포증식억제솔,계산IC50;아정등(AO)/추화을정(EB)형광염색법측정황기다당대MCF-7세포유도조망작용;응용류식세포의분석황기다당대MCF-7세포조망화세포주기적영향.결과 재급여2.5、5、10、20 mg/L황기다당48 h후,황기다당정농도의뢰성억제MCF-7세포적증식(r=0.985,P＜0.05),억제솔분별위(4.14±2.96)％、(7.14±2.10)％、(20.13±2.33)％、(64.66±5.15)％,고우공백대조조0％,단4개불동농도조적억제솔균저우양성대조조(90.31±4.92)％.황기다당48 h적IC50=16.83m/L.수착황기다당농도적축점증고,MCF-7세포중대표조망적마노색축점증다,세포핵취축、핵분렬,세포형태정현전형적조망특정.2.5、5、10、20 mg/L황기다당적조망솔분별위(2.37±0.98)％、(6.76±1.31)％、(11.65±1.46)％、(20.75±2.68)％,고우공백대조조(1.14±1.25)％(균P＜0.05),단균저우양성대조조(35.09±2.88)％(균P＜ 0.05).여공백대조조비교,황기다당이농도의뢰성유도MCF-7세포조망(r=0.991,P＜0.05),수농도적증가,세포조망솔승고,단균저우양성대조조.황기다당수농도적증가촉사S기세포비례축점승고,단저우공백화양성대조조(균P＜0.05),사처우G0-G1기세포적비례축점감소,잉고우공백화양성대조조(균P＜0.05).결론 황기다당억제인유선암MCF-7세포증식병유도기조망,사MCF-7세포생장증식정체재S기.
Objective To explore the in vitro anti-breast cancer activity of a traditional Chinese medicine,Astragalus polysaccharides (APS),in MCF-7 cells.Methods The MCF-7 cells were divided into three groups:treated with different concentrations of APS (2.5,5,10 and 20 mg/L) (APS group),with 10 μmol/L cisplastin (positive control group),with equivalent volume of media (blank control group).After 48 h,methylthiazolyl tetrazolium (MTT)test was used to evaluate the proliferation inhibition and then the IC50 was calculated; acridine orange (AO)-ethidium bromide (EB) fluorescence staining was used to evaluate the induced apoptosis of MCF-7 cells; flow cytometry was used to analyze the apoptosis and cell cycle.Results After 48 h treatment,different concentrations of APS (2.5,5,10 and 20 mg/L) were shown to inhibit the proliferation of MCF-7 cells in a dose-dependent manner (r=0.985,P＜0.05),with the rates of inhibition being (4.14±2.96)％,(7.14±2.10)％,(20.13±2.33)％ and (64.66±5.15)％,respectively.These inhibition rates were significantly higher than that in blank control group (0％),but lower than that in positive control group (90.31±4.92)％.The IC50 of APS at 48 h was 16.83 mg/L.Along with higher concentrations of APS,the MCF-7 cells showed more apoptosis as reflected by an increase in brown-stained cells and typical morphology of apoptosis such as karyopyknosis.The rates of apoptosis by APS were (2.37±0.98)％,(6.76±1.31)％,(11.65±1.46)％,(20.75±2.68)％,respectively,according different levels(2.5,5,10 and 20 mg/L).These rates were higher than that in blank control group (1.14± 1.25)％,but lower than that in positive control group (35.09±2.88) ％ (all P＜0.05).Compared with blank control group,APS induced apoptosis of MCF-7 cells in a dose-dependent manner (r=0.991,P＜0.05).The rates of apoptosis increased with higher doses of APS,but were all lower than that in the positive control group.The proportion of S-phase cells increased with higher concentration of APS,but was nevertheless lower than those in blank and positive control groups (all P＜0.05),whereas the proportion of G0-G1 phase cells decreased with higher concentration of APS,but was higher than those in blank and positive control groups (all P＜0.05).Conclusion APS is effective in inhibiting proliferation and inducing apoptosis of human breast cancer MCF-7 cells by arresting the cells at S phase.
