目的 研究补骨脂酚对人乳腺癌MCF-7细胞生长抑制作用及其机制。方法 采用噻唑蓝法（MTT法）考察补骨脂酚对MCF-7细胞生长的影响；采用流式细胞术检测补骨脂酚处理后细胞周期分布情况及活性氧（ROS）的产生；利用荧光显微镜观察补骨脂酚对MCF-7细胞核的影响；采用蛋白免疫印迹法检测补骨脂酚作用下MCF-7细胞凋亡、周期相关蛋白和丝裂原活化蛋白激酶（MAPK）家族蛋白的表达；引入ROS清除剂及MAPK家族蛋白抑制剂考察其在补骨脂酚作用下对MCF-7细胞生长抑制率和细胞周期相关蛋白表达的影响。结果 补骨脂酚可以呈时间、剂量依赖性抑制MCF-7细胞生长，其抑制作用明显强于阳性药5-氟尿嘧啶。补骨脂酚可诱导MCF-7细胞产生ROS，同时上调p-p53及p21蛋白表达水平，下调细胞周期相关蛋白CDK2和CyclinA2表达水平，进而引起细胞发生S期阻滞。而上述影响可被ROS清除剂Tiron逆转，表明ROS参与补骨脂酚诱导的S期阻滞。加入p38MAPK抑制剂SB203580后，细胞产生的ROS水平降低，表明ROS的产生依赖于p38MAPK途径。结论 补骨脂酚抑制MCF-7细胞增殖的作用与其引起细胞发生S期阻滞有关，ROS参与S期阻滞过程。

Objective To investigate the cell growth inhibitory effect and molecular mechanism of bakuchiol against human breast cancer MCF-7 cells. Methods The growth inhibitory effect of bakuchiol on MCF-7 cells was tested by MTT assay. Flow cytometry was used to investigate the distribution of cell cycle and ROS generation. Fluorescence microscope was used to observe the change of cell nucleus. Western blotting was used to detect the expression of the protein related to cell cycle and MAPK family. The ROS scavenger and inhibitors of MAPK family were introduced to investigate the effect on the growth inhibitory rate and the levels of cell cycle related protein by bakuchiol. Results Bakuchiol inhibited the cell growth on the MCF-7 cells in dose- and time-dependent manner, which showed stronger effect than that of 5-fluorouracil. Furthermore, bakuchiol induced S-phase arrest in MCF-7 cells via ROS generation. The production of ROS up-regulated p-p53 and p21 expression, and then decreased CDK2 and CyclinA2. The changes of bakuchiol on these proteins could be reversed by the ROS scavenger Trion, indicating that ROS was associated with bakuchiol-induced S-phase arrest. In addition, pretreatment with p38MAPK inhibitor SB203580 decreased bakuchiol-caused ROS generation, suggesting that the production of ROS was dependent on p38MAPK pathway. Conclusion The proliferation inhibitory effect of bakuchiol on MCF-7 cells is related with S-phase cell cycle arrest, and ROS plays a role in the bakuchiol-induced S-phase arrest.

R285.5