摘要
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 explore the effect of galangin on the migration and invasion of breast cancer and its action mechanism. Methods MTT assay was adopted to observe the effect of galangin at different concentration on the growth capacity of breast cancer cells MDA-MB-231. Wound healing and Transwell assay were conducted to detect the effect of galangin on the migration and invasion of MDA-MB-231 cells. Western blotting and immunofluorescence assay were adopted to investigate the migration and invasion related proteins including PI3 K, AKT, MMP-2, and MMP-9. The effect of galangin on NF-κb p65 phosphorylation and NF-κb p65 nuclear transition in vitro was assayed by Western blotting and immunofluorescence assay. Furthermore, the antitumor effect of galangin in MDA-MB-231 tumor bearing BALB/c mice model was assayed in vivo. Results Galangin inhibited the migration of MDAMB-231 cells in a dose-dependent manner. In wound healing and transwell experiments, with the increase in the concentration of galangin, the number of migrant MDA-MB-231 cells and the invasion capacity of breast cancer cells decreased. Galangin could decrease the protein expression of PI3 K, Akt, MMP-2, and MMP-9. Besides, galangin significantly inhibit the phosphorylation of transcription factor NF-κB p65 and NF-κB p65 nuclear transition. Galangin also inhibited the growth of MDA-MB-231 cancer cells in vivo and down-regulated PI3 K, Akt, MMP-2, and MMP-9. Conclusion Galangin could inhibit the growth capacity of breast cancer cells MDA-MB-231 and block the migration and invasion of MDA-MB-231 cells. The mechanism for antitumor effect of galangin is through inhibiting NF-κb p65 activity and MMP-2/9 expression, and blocking PI3K-AKT signaling pathway.
出处
《中草药》
CAS
CSCD
北大核心
2016年第10期1731-1739,共9页
Chinese Traditional and Herbal Drugs
基金
江苏省卫生厅医学科研项目(Z201304)
作者简介
许奕夫，男，学士，主要从事医院药学研究。E-mail：xyfsuzhou@163．com
通信作者姚鑫，男，硕士，主要从事医院药学研究。E-mail：yaobest@163．com
