复方苦参联合5-氟尿嘧啶调控细胞周期蛋白D1抑制乳腺癌生长作用的研究

Effectiveness Study of Compound Sophora Flavescens Combined with 5-Fluorouracil on Inhibiting the Growth of Breast Cancer by Regulating Cyclin D1

为了解苦参联合5-氟尿嘧啶(5-Fu)对乳腺癌MCF-7细胞增殖和凋亡及对Cyclin D1表达的影响,采用流式细胞术观察联合用药组(复方苦参+5-Fu)和对照组(二甲基亚砜DMSO+5-Fu)处理后MCF-7细胞凋亡的情况;采用免疫印迹法(Western Blot)观察两组MCF-7细胞Cyclin D1的表达;采用CCK-8细胞增殖实验观察两组MCF-7细胞增殖情况;采用RT-PCR观察两组MCF-7细胞中Cyclin D1 mRNA的表达;建立裸鼠荷瘤模型,并观察两组肿瘤组织中Cyclin D1的表达.结果联合用药组处理的MCF-7细胞凋亡(12.7%)高于对照组(6.2%),差异显著.联合用药组MCF-7细胞中Cyclin D1 mRNA及Cyclin D1的蛋白表达均低于对照组;联合用药组MCF-7细胞的增殖能力低于对照组(P<0.05);裸鼠荷瘤后,联合用药组的直径为(79.8±10.3)mm,明显小于对照组的直径(176.7±26.2)mm(P<0.05).说明苦参可明显抑制乳腺癌MCF-7细胞中Cyclin D1的表达,并抑制MCF-7细胞的增殖,加速MCF-7细胞的凋亡,其机制可能与抑制Cyclin D1的表达有关.

To explore the influence of compound sophora flavescens 5-Fu on proliferation and apoptosis of MCF-7 breast cancer cells and the expression of Cyclin D1,flow cytometry was used to observe the apoptosis of MCF-7 cells treated by the combined groups（ compound sophora flavescens 5-Fu） or the control group（ DMSO 5-Fu）. Western Blot was used to observe the expression of Cyclin D1 in the two groups. CCK-8 Cell proliferation experiment was conducted to observe the proliferation of MCF-7 cells,and RT PCR was used to detect Cyclin D1 mRNA expression of MCF-7 cells. The tumor model of nude mice was established to compare the expression of Cyclin D1 of tumor tissues in the two groups. Our results showed that the apoptosis of MCF-7 cells（ 12. 7%） in the combined treatment group was higher than that in the control group（ 6.2%）. The expression of Cyclin D1 mRNA and Cyclin D1 protein of MCF-7 cells in the combined treatment group was lower than that in the control group. The proliferation capacity of MCF-7 cells in the combined treatment group was also lower than that in the control group（ P〈0.05）. In MCF-7 tumor-burdened nude mice,the diameter of the biggest tumor of the combined treatment group（ 79.8±10.3） mm was substantially smaller than that of the control group（ 176.7 ± 26.2） mm. The difference was statistically significant（ P〈 0. 05）. It indicates that compound sophora flavescens can significantly inhibit the proliferation of MCF-7 cells and increase the apoptosis of MCF-7 cells,which may be related to the inhibition of Cyclin D1 expression.