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目的:研究蛇床子素配伍补骨脂素对人乳腺癌高骨转移细胞株MDA-MB-231BO增殖与侵袭的影响,并探讨其分子机制。方法:Cell Counting Kit-8(CCK-8)测定蛇床子素、补骨脂素对细胞的毒性大小。根据L_9(3~4)正交设计表,Transwell侵袭实验筛选最佳配伍浓度。Western blot及real-time PCR检测蛇床子素配伍补骨脂素对Smad2、Smad3、Smad4、Smad7表达的影响。结果:蛇床子素和补骨脂素都能显著抑制细胞的增殖活性,且存在剂量依赖性。150μM的蛇床子素配伍100μM的补骨脂素时细胞侵袭率最低。Western b1ot和real-timePCR结果显示最佳配伍组Smad2、Smad3、Smand4蛋白及mRNA表达量明显降低,Smad7蛋白及mRNA表达量明显增加。结论:蛇床子素和补骨脂素配伍使用对细胞的侵袭抑制作用优于两个单体单用效果。配伍组通过干预TGF-β/Smads信号传导通路的各个环节从而抑制MDA-MB-231BO的增殖与侵袭。
Objective: To investigate the effects of osthole and psoralen on proliferation and invasion of human breast cancer cell line MDA-MB-231BO and its molecular mechanism. Methods: Cell Counting Kit-8 (CCK-8) was used to determine the cytotoxicity of osthole and psoralen to cells. According to L_9 (3 ~ 4) orthogonal design table, Transwell invasion experiment screening the best compatibility concentration. The expression of Smad2, Smad3, Smad4 and Smad7 were detected by Western blot and real-time PCR in combination with osthole and psoralen. Results: Both osthole and psoralen inhibited the cell proliferative activity in a dose - dependent manner. 150μM ostholol with 100μM psoralen at the lowest cell invasion rate. Western b1ot and real-time PCR results showed that the best compatibility group Smad2, Smad3, Smand4 protein and mRNA expression was significantly reduced, Smad7 protein and mRNA expression increased significantly. Conclusion: The inhibitory effect of osthole and psoralen on invasion of cells is better than that of two monomers alone. The compatibility group inhibits the proliferation and invasion of MDA-MB-231BO by intervening in all aspects of TGF-β / Smads signaling pathway.