阿西替尼对乳腺癌细胞生物学活性的影响

Effects of axitinib on biological activities of breast cancer cells

目的研究阿西替尼对乳腺癌细胞MCF-7增殖、凋亡和迁移的影响。方法 MTT法检测阿西替尼对MCF-7细胞增殖的影响,流式细胞仪检测阿西替尼作用MCF-7细胞48 h后对细胞周期的影响,并进一步用免疫印迹的方法检测周期蛋白Cyclin D和Cyclin E表达量的变化;流式细胞仪检测阿西替尼对MCF-7细胞凋亡的影响,并用免疫印迹的方法检测凋亡相关蛋白Pro-Caspase-8和Pro-Caspase-9表达水平的变化;Transwell小室实验检测阿西替尼对MCF-7细胞迁移的影响,逆转录-聚合酶链反应(RT-PCR)的方法检测迁移相关蛋白MMP-2和MMP-9表达的变化。结果阿西替尼抑制MCF-7细胞增殖呈时间和剂量依赖性,将MCF-7细胞抑制在G1期,并且G1期调控蛋白随阿西替尼浓度增加表达量逐渐降低;阿西替尼促进MCF-7细胞的凋亡,并且蛋白前体Pro-Caspase-8和Pro-Caspase-9的表达量随阿西替尼浓度的增加而逐渐减少;阿西替尼抑制MCF-7细胞的迁移,迁移相关蛋白MMP-2和MMP-9的m RNA水平随阿西替尼浓度增加而表达下调。结论阿西替尼通过把MCF-7细胞控制在G1期而抑制细胞的增殖,阿西替尼促进MCF-7细胞的凋亡,并能抑制其转移。

【Objective】 To explore the effect of axitinib on proliferation, apoptosis and migration of Michigan Cancer Foundation-7（MCF-7） cells. 【Methods】 The effect of axitinib on cell proliferation was measured by MTT colorimetry. The expression levels of cyclin D and cyclin E proteins were detected by Western blot. The effect of axitinib on cell apoptosis was analyzed using flow cytometry by double staining with annexin V and propidium iodide. The expression levels of pro-Caspase-8 and pro-Caspase-9 proteins associated with apoptosis were detected by Western blot. The invasion potential was evaluated by Transwell assay, and the expression levels of MMP-2 and MMP-9 proteins were measured by RT-PCR. 【Results】 The MCF-7 cell proliferation, apoptosis and invasion were depressed by axitinib in a time or dose-dependent manner. And the levels of associated proteins such as cyclin D, cyclin E, pro-Caspase-8, pro-Caspase-9 and MMP-2, MMP-9m RNA became lower and lower as the axitinib concentration increased.【Conclusion】 Axitinib represses MCF-7 cell proliferation by controlling the cells in G1 stage, triggers cell apoptosis and decreases its invasion potential.