FAK对黑米花青素抑制HER-2阳性乳腺癌转移的影响研究

Role of focal adhesion kinase in black rice anthocyanins inhibiting HER-2 positive human breast cancer cells

目的:探讨粘着斑激酶(Focal Adhesion Kinase,FAK)在黑米花青素(Black Rice Anthocyanins,BRACs)抑制人表皮生长因子受体2(Human epidermal growth factor-2;HER-2)阳性乳腺癌转移中的作用。方法:以正常永生化乳腺细胞MCF-10A、HER-2阴性乳腺癌细胞MCF-7、HER-2阳性乳腺癌细胞MDA-MB-453为研究对象,使用Cell Counting Kits-8(CCK8)试剂盒检测细胞活性;伤痕愈合实验检测细胞转移状态;免疫印迹实验测定FAK、c Src表达及磷酸化水平;免疫共沉淀实验检测HER/FAK蛋白间相互作用。结果:与对照组相比,分别用50、100、200、400μg/m L BRACs处理MDA-MB-453细胞后,抑制率分别为18.74%、21.06%、22.82%、40.12%;200μg/m L BRACs处理后MDA-MB-453细胞迁移距离减少37%;FAK及c Src磷酸化水平降低,HER-2/FAK蛋白间相互作用减弱。结论:BRACs通过减弱FAK与HER-2受体相互作用抑制FAK及c Src磷酸化从而抑制MDA-MB-453细胞增殖和转移,这可能是黑米花青素抑制MDA-MB-453细胞增殖和转移的分子机制之一。

Objective： To explore the role of focal adhesion kinase in HER-2 positive human breast cancer cells antimetastasis by black rice anthocyanins（ BRACs）. Methods： To observe the effect of BRACs on the proliferation of MCF-10 A,MCF-2 and MDA- MB-453 cells with CCK8 method. Metastasis of MCF-10 A,MCF-2 and MDA- MB-453 cells were detected by wound healing experiments.The level of total and phosphorylated FAK and c Src were detected by western blot.The interaction between HER-2 and FAK were analyzed by immunoprecipitation Results：Different concentration of BRACs including 50,100,200,400 μg / m L were used to detect the influence which BRACs play on the growth MDA- MB- 453 cells. Compared with control,the inhibition ratio of each group were 18.74%,21.06%,22.82%,40.12% respectively. The migration distance of MDA- MB- 453 cells was decreased 37% by200 μg / m L BRACs. The level of phosphorylated FAK and c Src were reduced by BRACs and the interaction between HER- 2 / FAK was also decreased by BRACs. Conclusion： BRACs suppress the metastasis of HER- 2positive breast cancer cells via inhibiting the phosphorylation of FAK and c Src by decrease the interaction between FAK and HER-2 which maybe a mechanism of anti- metastasis by BRACs.