蛇床子素体外抗乳腺癌活性研究

In vitro research of anti-breast cancer activity of Osthole

目的:探索蛇床子素对具有HER-2/neu高表达的人乳腺癌SK-BR-3和MDA-MB-231细胞系中的促血管生成因子的表达影响,并阐述其可能的活性机制。方法:以蛇床子素与表皮生长因子(EGF)对具有HER-2/neu高表达的人乳腺癌SK-BR-3和MDA-MB-231细胞系进行单独或联合处理,用酶联免疫吸附测定法(ELISA)定量测定肿瘤细胞促血管生成因子(VEGF、MMP-9)的表达,Western blot检测HER-2/neu及MAPK信号通路关键蛋白ERK1/2的磷酸化水平改变,QRT-PCR检测VEGF、MMP-9在m RNA水平的变化。结果:蛇床子素对HER-2/neu高表达的人乳腺癌SK-BR-3和MDA-MB-231细胞系均具有显著的增殖抑制作用。EGF能显著促进HER-2/neu高表达的人乳腺癌SK-BR-3和MDA-MB-231细胞系表达肿瘤促血管生成因子,蛇床子素能显著能显著抑制ERK1/2蛋白的磷酸化,下调经EGF处理后肿瘤促血管生成因子(VEGF、MMP-9)的表达,并在m RNA水平显著抑制促血管生成因子的表达。结论:蛇床子素可能通过抑制HER-2/neu活性而阻断其下游的MAPK信号通路,通过抑制了ERK的磷酸化减少了MMP-9的表达,从而抑制乳腺癌细胞分泌VEGF,实现其对促血管生成因子表达水平的抑制作用。

Objective:To explore the effect of osthole on the expression of pro-angiogenic factors in human breast cancer SK-BR-3and MDA-MB-231 cell lines with high HER-2/neu expression and to elucidate its possible mechanism.Method:Human breast cancer SK-BR-3 and MDA-MB-231 cell lines with high HER-2/neu expression were treated with osthole and epidermal growth factor(EGF)either alone or in combination.Enzyme-linked immunosorbent assay(ELISA)was used to quantitatively determine the expression of tumor angiogenic factors(VEGF,MMP-9)and QRT-PCR detection of VEGF,MMP-9 changes in mRNA levels.Western blot was then applied to verify the change of phosphorylation of ERK-1/-2.Result:Osthole could significantly inhibit the proliferation of human breast cancer cell line SK-BR-3 and MDA-MB-231 with high HER-2/neu expression.EGF can significantly promote the expression of tumor angiogenic factors in human breast cancer SK-BR-3 and MDA-MB-231 cells with high HER-2/neu expression.Osthole could significantly inhibit the phosphorylation level of ERK-1/-2,reduce the expression of tumor angiogenic factors(VEGF,MMP-9)after EGF treatment and significantly inhibit the expression of pro-angiogenic factors at the mRNA level.Conclusion:Osthole could possibly block the downstream of MAPK signaling pathway by inhibiting the activity of HER-2/neu and reduce the expression of MMP-9 by inhibiting the phosphorylation of ERK,thereby inhibiting the secretion of VEGF from breast cancer cells and promoting its effect on pro-angiogenic inhibition.