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目的探究高良姜素对MDA-MB-231乳腺癌细胞迁移及侵袭的影响及其作用机制。方法采用MTT法观察不同浓度高良姜素对乳腺癌细胞MDA-MB-231增殖的影响;划痕愈合实验和Transwell小室迁移实验考察高良姜素对MDA-MB-231水平迁移能力和垂直迁移能力的影响;Transwell小室侵袭实验分析高良姜素对MDA-MB-231细胞侵袭能力的影响;采用蛋白免疫印迹(Western blotting)法考察高良姜素对MDA-MB-231细胞PI3K、Akt、MMP-2、MMP-9蛋白表达的影响,免疫荧光染色实验验证高良姜素对MDA-MB-231细胞PI3K、Akt、MMP-2、MMP-9蛋白表达的影响;Western blotting法和免疫荧光染色实验考察高良姜素对核转录因子(NF-κB)p65磷酸化表达和入核的影响;制备乳腺癌MDA-MB-231细胞株裸鼠移植瘤模型,高良姜素200μg/kg ig给药15 d,检测肿瘤体积和质量,计算抑瘤率,Western blotting分析肿瘤组织PI3K、AKT、MMP-2、MMP-9表达及NF-κB p65磷酸化表达情况。结果高良姜素体外浓度为50、100、200μmol/L时可显著抑制MDA-MB-231细胞的增殖;高良姜素能剂量依赖性抑制PI3K、Akt、MMP-2、MMP-9蛋白表达,降低NF-κB p65磷酸化表达和入核;200μg/kg高良姜素能抑制裸鼠乳腺癌的生长,抑瘤率为43.66%;高良姜素能抑制瘤组织中PI3K、Akt、MMP-2、MMP-9表达以及NF-κB p65磷酸化表达。结论高良姜素在体内对MDA-MB-231生长有明显的抑制作用,在体外能抑制MDA-MB-231细胞增殖、迁移、侵袭,抑制相关蛋白PI3K、Akt和MMP-2、MMP-9蛋白表达,抑制转录因子NF-κB磷酸化表达及入核,阻断其激活。
Objective To investigate the effect of galangin on the migration and invasion of MDA-MB-231 breast cancer cells and its mechanism. Methods The effects of galangin at different concentrations on the proliferation of breast cancer cells MDA-MB-231 were observed by MTT assay. Scratch healing assay and Transwell chamber assay were used to investigate the effects of galangin on the migration and migration of MDA-MB-231 The effect of galangin on the invasiveness of MDA-MB-231 cells was analyzed by Transwell chamber invasion assay. The effects of galangin on the expression of PI3K, Akt, MMP-2, The effect of galangin on the protein expression of PI3K, Akt, MMP-2 and MMP-9 in MDA-MB-231 cells was verified by immunofluorescence staining. The results of Western blotting and immunofluorescence staining showed that galangin On the phosphorylation and nuclear translocation of nuclear factor-kappa B (NF-κB) p65.Methods The nude mice model of breast cancer MDA-MB-231 cells was established and treated with galangin 200 μg / kg for 15 days. Volume and quality of the tumor were calculated. The tumor inhibition rate was calculated. The expression of PI3K, AKT, MMP-2, MMP-9 and the phosphorylation of NF-κB p65 were detected by Western blotting. Results galangin significantly inhibited the proliferation of MDA-MB-231 cells in vitro at concentrations of 50,100 and 200μmol / L. Galangin could inhibit the expression of PI3K, Akt, MMP-2 and MMP-9 in a dose- κB p65 phosphorylation and nuclear entry; galangin 200μg / kg can inhibit the growth of breast cancer in nude mice inhibition rate was 43.66%; galangin can inhibit the expression of PI3K, Akt, MMP-2, MMP-9 and NF- κB p65 phosphorylation. CONCLUSION Galangin can significantly inhibit the growth of MDA-MB-231 cells in vitro and inhibit the proliferation, migration and invasion of MDA-MB-231 cells in vitro and inhibit the expression of PI3K, Akt and MMP-2, MMP- Expression, inhibition of transcription factor NF-κB phosphorylation and nuclear entry, blocking its activation.