中药提取物配伍对乳腺癌MDA-MB-231BO细胞侵袭作用的影响及其机制

目的：探讨中药蛇床子、补骨脂、附子有效成分蛇床子素、补骨脂素、乌头碱对乳腺癌细胞株MDA-MB-231BO的侵袭抑制作用及其机制。方法：采用均匀设计实验方案进行组方设计,逐步回归分析拟合蛇床子素、补骨脂素配伍乌头碱的回归方程;采用人乳腺癌骨高转移细胞（MDA-MB-231BO）体外侵袭实验,实时荧光定量聚合酶链反应检测,观察药物处理MDA-MB-231BO细胞后,细胞侵袭作用特点和相关基因的表达。结果：通过逐步回归方程,获取蛇床子素、补骨脂素和乌头碱3个药物的最佳配伍比例为6.44∶8.89∶9.44;有效成分和阳性药物唑来磷酸注射液在体外作用24h可以显著抑制MDA-MB-231BO细胞的侵袭;药物处理组转化生长因子β1（transforming growth factor-β1,TGF-β1）、Smad2、Smad3、Smad4、Smad7、核因子κB（nuclear factor-κB,NF-κB）、NF-κB受体活化因子（receptor activator of NF-κBligand,RANK）mRNA表达量显著降低,最佳药物配伍组和唑来磷酸药物组与空白对照组,最佳药物配伍组与最差药物配伍相比差异有统计学意义（P〈0.01）。结论：蛇床子素、补骨脂素、乌头碱配伍应用可显著抑制MDA-MB-231BO细胞侵袭,其机制与抑制TGF-β/Smad的信号转导有关,同时可降低NF-κB和RANK的表达。

Inhibitory effects of osthole, psoralen and aconitine on invasive activities of breast cancer MDA-MB-231BO cell line and the mechanisms

Objective：To explore the inhibitory effects and to investigate the mechanisms of combined treatment of osthole,psoralen with aconitine on human breast cancer cell line MDA-MB-231BO.Methods：The best inhibitory concentration of osthole,psoralen combined with aconitine on MDA-MB-231BO cells was obtained by stepwise regression analysis after adopting a uniform experiment design.The invasive activities were observed by transwell assays,and expressions of transforming growth factor-β1（TGF-β1）,Smad2,Smad3,Smad4,Smad7,nuclear factor-κB（NF-κB）and receptor activator of NF-κB ligand（RANK）mRNAs were analyzed by real-time quantitative polymerase chain reaction.Results：The optimal combination concentrations of osthol,psoralen and aconitine were 6.44,8.89 and 9.44 μg/mL,respectively.Cell invasion was significantly inhibited after 24 hours of treatment using the combination drugs and zoledronic acid.TGF-β1,Smad2,Smad3,Smad4,Smad7,NF-κB and RANK mRNA expressions of the optimal combination group and zoledronic acid group were significantly reduced compared with the control group（P0.01）.Furthermore,TGF-β1,Smad2,Smad3,Smad4,Smad7,NF-κB and RANK mRNA expressions of the optimal combination group were significantly lower than those of the weak combination group（P0.01）.Conclusion：Combination treatment of osthole,psoralen with aconitine can inhibit cancer cell invasion,which is a result of alteration of the TGF-β/Smad signaling pathway and down-regulation of NF-κB and RANK expressions.