氨氯地平对人乳腺癌细胞MCF-7细胞的抑制作用及机制研究

目的观察氨氯地平对人乳腺癌细胞MCF-7周期、周期蛋白相关基因及产物表达的影响,探讨氨氯地平对人乳腺癌MCF-7细胞周期的影响及其机制。方法MTT检测细胞增殖;流式细胞仪分析细胞周期;RT-PCR技术检测细胞周期相关基因cyclinD1、p21mRNA的表达;Western blot检测细胞周期蛋白cyclinD1、p21的蛋白表达。结果氨氯地平剂量和时间依赖性的抑制人乳腺癌MCF-7细胞增殖,IC50为14.439μmol.L-1。经7.22μmol.L-1(1/2IC50)、14.439μmol.L-1(IC50)、28.88μmol.L-1(2IC50)的氨氯地平作用人乳腺癌MCF-7细胞48h,G0/G1期细胞较对照组明显增高(P<0.05);并使人乳腺癌MCF-7细胞中cyclinD1mRNA及蛋白表达降低;p21mRNA及蛋白表达升高。结论氨氯地平对人乳腺癌MCF-7细胞具有抗增殖作用,并使细胞阻滞于G1期。其G1阻滞机制可能与调控细胞周期相关基因cyclinD1、p21mRNA及蛋白的表达相关。

Study on inhibitory effects and mechanism of amlodipine on human breast carcinoma cell line MCF-7

Aim To observe the effects of amlodipine on cell cycle,cell cycle-related genes and cyclin expression of human breast carcinoma MCF-7 cells.And to probe effects of amlodipine on cell cycle and its mechanism of human breast carcinoma MCF-7 cells.Methods In vitro growth inhibitory effects of amlodip-ine on human breast carcinoma MCF-7 cells were determined by MTT assay,cell cycle distribution was detected by flow cytometry,the Mrna expression levels of cell cycle-related genes cyclinD1 and p21 were treated by RT-PCR,and the protein expression of cell cycle protein cyclinD1 and p21 were assessed by Western blot.Results With dose and time dependently,amlodipine could inhibit the proliferation of human breast carcinoma MCF-7 cells in vitro.The IC_(50) was 14.439 μmol·L~(-1).When breast cancer cells MCF-7 were treated with 7.22 μmol·L~(-1)(1/2 IC_(50)),14.439 μmol·L~(-1)(IC_(50))and 28.88 μmol·L~(-1)(2IC_(50))amlodipine for 48 h,the ratios in the G_0/G_1 phase were significantly increased as compared with control group(P<0.05).Amlodipine inhibited the expression of Mrna and protein of cyclinD1,while increased the expression of Mrna and protein of p21.Conclusions Amlodipine exhibits obvious anti-tumor activities on human breast carcinoma MCF-7 cells and arrest cell cycle in G_1 phase.The mechanism of G_1 phase arresting may be related to modulating the Mrna and protein expression of cell cycle-related gene cyclinD1 and p21.