大蒜素逆转人乳腺癌MCF-7细胞顺铂耐药性的研究

目的]探索大蒜素对人乳腺癌MCF-7细胞顺铂耐药性的逆转作用及调控机制。方法]以对数生长期人乳腺癌耐顺铂MCF-7/DDP细胞为受试细胞,设空白对照组(DMSO)、大蒜素(20μg/mL)单药组、顺铂(6μg/mL)单药组、联合组(大蒜素20μg/mL+顺铂6μg/mL)。药物干预48 h后,噻唑蓝(MTT)染色法检测MCF-7/DDP细胞增殖抑制率,应用Annexin V-FITC染色、碘化丙啶(PI)染色流式细胞术检测细胞凋亡和分析细胞周期分布,Western blot法检测细胞凋亡相关调控蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)]和细胞周期相关调控蛋白特异性周期蛋白-D1(Cyclin D1),周期蛋白依赖激酶2、4、6(CDK2、CDK4、CDK6)]的表达。结果]与顺铂单药组比较,联合组MCF-7/DDP细胞增殖抑制率、凋亡率升高,G0/G1期MCF-7/DDP细胞比例升高,Bax表达量升高而Bcl-2表达量降低、Bax/bcl-2值升高,Cyclin D1、CDK2、CDK4、CDK6表达量降低,差异均具有统计学意义(P<0.05或P<0.01)。与空白对照组比较,大蒜素单药组MCF-7/DDP细胞G0/G1期比例升高,Bax表达量升高且Bcl-2表达量降低、Bax/bcl-2值升高,Cyclin D1、CDK2、CDK4、CDK6表达量降低,差异均具有统计学意义(P<0.05或P<0.01)。结论]大蒜素具有逆转人乳腺癌MCF-7细胞顺铂耐药性的药理学作用,可能与调节细胞凋亡、周期相关蛋白表达进而诱导MCF-7/DDP细胞凋亡、阻滞其细胞周期进程有关。

Study of Allicin reverses Cisplatin resistance in human breast cancer MCF-7 cells

Objective]To study the effect of Allicin reverses Cisplatin resistance in human breast cancer MCF-7 cells and related regulatory mechanisms.Methods]Taking human cisplatin-resistant MCF-7/DDP cells in logarithmic growth phase as test cells,set blank control group(DMSO),Allicin group(20μg/mL),Cisplatin group(6μg/mL)and combined group(Allicin 20μg/mL+Cisplatin 6μg/mL).48h after given drugs,the proliferation inhibition rate of MCF-7/DDP cells was detected by MTT method,the apoptosis and analysis of cell cycle distribution were detected by Annexin V-FITC staining and PI staining flow cytometry,the expression of Bcl-2,Bax,Cyclin D1,CDK2,CDK4,CDK6 proteins were detected by Western blot.Results]Compared with Cisplatin group,the proliferation inhibition rate and apoptosis rate of human breast cancer MCF-7 cells in combination group decreased,the ratio of the cells in G0/G1 phase increased,the expression of Bax increased and the expression of Bcl-2 decreased the Bax/bcl-2 ratio increased,the expression of Cyclin D1,CDK2,CDK4,CDK6 proteins decreased,all of the difference above were significant(P<0.05 or P<0.01).Compared with blank group group,the ratio of the cells in G0/G1 phase of Crocin group increased,the expression of Baxincreased and the expression of Bcl-2 decreased,the Bax/bcl-2 ratio increased,the expression of Cyclin D1,CDK2,CDK4,CDK6 proteins decreased,all of the difference above were significant(P<0.05 or P<0.01).Conclusions]Allicin has pharmacological effects on reversing Cisplatin resistance in human breast cancer MCF-7 cells,and its mechanism may be related to the regulation of apoptosis-related and cycle-related proteins expression,promoting MCF-7/DDP cell apoptosis and blocking the cell cycle process.