基于ERK/NF-κB通路探讨苦杏仁苷对乳腺癌MDA-MB-231细胞增殖、侵袭、迁移的影响

Effects of amygdalin on proliferation,invasion and migration of breast cancer MDA-MB-231 cells based on ERK/NF-κB pathway

目的探讨苦杏仁苷对乳腺癌MDA-MB-231细胞增殖、侵袭、迁移及细胞外信号调节激酶(ERK)/核因子(NF)-κB通路的影响。方法将乳腺癌MDA-MB-231细胞随机分成对照组、紫杉醇组(10μg/ml)、苦杏仁苷低剂量组(20μg/ml)、苦杏仁苷高剂量组(40μg/ml);以上各组每孔设6个平行样,培养72 h。实验结束后,噻唑蓝(MTT)法测定乳腺癌MDA-MB-231细胞增殖水平,Transwell小室测定乳腺癌MDA-MB-231细胞侵袭迁移水平,实时荧光定量聚合酶链反应(PCR)及Western印迹法测定乳腺癌MDA-MB-231细胞ERK、NF-κB mRNA和蛋白表达水平。结果与对照组比较,紫杉醇组、苦杏仁苷低、高剂量组光密度(OD)值、细胞存活率、穿膜数、ERK、NF-κB mRNA和蛋白表达水平显著降低(P<0.05);与紫杉醇组比较,苦杏仁苷低剂量组上述指标显著升高(P<0.05),苦杏仁苷高剂量组上述指标无显著差异(P>0.05);与苦杏仁苷低剂量组比较,苦杏仁苷高剂量组上述指标显著降低(P<0.05)。结论苦杏仁苷能明显抑制乳腺癌MDA-MB-231细胞增殖、侵袭、迁移,其机制可能与苦杏仁苷抑制ERK、NF-κB mRNA和蛋白表达进而抑制ERK/NF-κB通路的激活有关。

Objective To investigate the effect of amygdalin on breast cancer MDA-MB-231 cells proliferation,invasion,migration and extracellular signal-regulated kinase(ERK)/nuclear factor(NF)-κB pathway.Methods Breast cancer MDA-MB-231 cells were randomly divided into control group,paclitaxel group(10μg/ml),amygdalin low dose group(20μg/ml),amygdalin high dose group(40μg/ml);6 parallel samples were set for each of the above groups and cultured for 72 h.After the experiment,the proliferation level of breast cancer MDA-MB-231 cells were measured by thiazolyl terazolium(MTT)method,the invasion and migration levels of breast cancer MDA-MB-231 cells were measured by Transwell chamber,real-time fluorescence quantitative polymerase chain reaction(PCR)and Western blotting were used to determine the mRNA and protein expression levels of ERK and NF-κB in breast cancer MDA-MB-231 cells.Results Compared with control group,the optical density(OD)value,cell viability,number of penetrating membrane,ERK,NF-κB mRNA and protein expression levels of paclitaxel group,amygdalin low dose group and amygdalin high dose group were significantly decreased(P<0.05);compared with paclitaxel group,the above indexes in amygdalin low dose group were significantly increased(P<0.05);there was no significant difference in the above indexes in amygdalin high dose group and paclitaxel group(P>0.05).Compared with amygdalin low dose group,the above indexes in amygdalin high dose group were significantly decreased(P<0.05).Conclusions Amygdalin could significantly inhibit the proliferation,invasion and migration of breast cancer MDA-MB-231 cells,and its mechanism might be related to that amygdalin could inhibit the expressions of ERK,NF-κB mRNA and protein,thereby inhibiting activation of ERK/NF-κB pathway.