和厚朴酚抑制人乳腺癌SK-BR-3细胞的生长及其机制

Honokiol inhibits the growth of human breast cancer SK-BR-3 cells and its underlying mechanism

目的:探讨和厚朴酚对人乳腺癌SK-BR-3细胞增殖、凋亡、侵袭和迁移的影响,及可能的作用机制。方法:用不同浓度的和厚朴酚处理人乳腺癌SK-BR-3细胞,分别采用结晶紫染色法和MTT法检测细胞的增殖能力,用FCM法检测和厚朴酚对细胞周期和凋亡率的影响,用划痕愈合实验和Transwell实验检测对细胞迁移和侵袭能力的影响。同时,采用蛋白质印迹法检测和厚朴酚处理对SK-BR-3细胞中增殖相关蛋白增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、细胞周期相关蛋白cyclin D1、凋亡相关蛋白(Bcl-xL、Bax、caspase3和cleaved-caspase3)以及细胞迁移和侵袭相关蛋白[基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)、波形蛋白(vimentin)和E-钙黏蛋白(E-cadherin)]表达水平的影响,对MAPK/ERK信号通路中总ERK以及磷酸化ERK蛋白的表达水平的影响。结果:与空白对照组相比,随着和厚朴酚浓度的提高,和厚朴酚对SKBR-3细胞的增殖、迁移和侵袭能力的抑制作用显著增强(P<0.01)。和厚朴酚处理后,SK-BR-3细胞中PCNA和cyclin D1蛋白的表达水平明显下降(P<0.01);抗凋亡蛋白Bcl-xL的表达水平明显下降(P<0.01),促凋亡蛋白Bax的表达水平明显升高(P<0.05),caspase3的表达水平明显下降(P<0.01)而cleaved-caspase3的蛋白水平上升(P<0.05);同时,迁移和侵袭相关蛋白MMP-2和vimentin的表达水平明显下降(P<0.01),而E-cadherin蛋白的表达水平明显升高(P<0.01);MAPK/ERK信号通路中p-ERK1/2的蛋白水平明显降低(P<0.01)。结论:和厚朴酚可以在诱导人乳腺癌SK-BR-3细胞凋亡同时抑制其增殖、迁移和侵袭,具体作用机制可能与抑制MAPK/ERK信号通路的活性有关。

Objective:To study the effects of honokiol on the proliferation,apoptosis,invasion and migration of human breast cancer SK-BR-3 cells and the underlying molecular mechanism.Methods:Human breast cancer SK-BR-3 cells were treated with different concentrations of honokiol,Then,the cell viability was assessed by crystal violet staining and MTT assay,the effect of honokiol on the cell cycle and apoptosis of SK-BR-3 cells was assessed by FCM assay,and the effect of honokiol on the migration and invasion of SK-BR-3 cells was evaluated by wound healing assay and Transwell assay.In the meantime,the effect of honokiol on the expression levels of proliferation associated protein PCNA(proliferating cell nuclear antigen),cell cycle associated protein cyclin D1,apoptosis associated proteins(Bcl-xL、Bax、caspase3 and cleaved-caspase3),cell migration and invasion associated proteins[MMP-2(matrix metalloproteinase-2),vimentin and E-cadherin],as well as ERK and phosphorylated ERK of the MAPK/ERK signaling pathway was examined by Western blotting.Results:Compared with blank control,honokiol treatment significantly inhibited the proliferation,migration and invasion of SK-BR-3 cells as the treatment concentration increased(P<0.01).After honokiol treatment in SK-BR-3 cells,the expression levels of PCNA and cyclin D1 decreased significantly(P<0.01);the expression levels of anti-apoptotic protein Bax increased obviouly(P<0.05),the expression level of caspase3 decreased significantly(P<0.01)while that of cleaved-caspase3 increased(P<0.05);the expression levels of migration and invasion assaociated proteins MMP-2 and vimentin decreased evidently(P<0.01)while taht of E-cadherin increased significantly(P<0.01);the expression level of p-ERK1/2 of the MAPK/ERK signaling pathway decreased significantly(P<0.01).Conclusion:Honokiol can induce the apoptosis while inhibit the proliferation,migration and invasion of human breast cancer SK-BR-3 cells.The potential mechanism may be related to the inhibition of the activity of the MAPK/ERK sig naling pathway.