大蒜素逆转人乳腺癌MCF-7细胞顺铂耐药性的研究

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

关键词:  大蒜素  乳腺癌  顺铂  耐药  细胞周期

Key words:  Allicin  breast cancer  Cisplatin  resistance  cell cycle

摘要:

[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（20 μg/mL）单药组、顺铂（6 μg/mL）单药组、联合组（大蒜素20 μg/mL+顺铂6 μg/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。

Abstract:

[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 (6 μg/mL) and combined group (Allicin 20 μg/mL +Cisplatin 6 μg/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.