基于CYP19探讨雷公藤甲素抗ER(+)人乳腺癌MCF-7细胞作用机理

摘要

目的 基于CYP19基因阐明雷公藤甲素抗ER(+)人乳腺癌细胞的作用机制.方法 雷公藤甲素组与来曲唑组相对照,采用MTT法观察雷公藤甲素对MCF-7细胞增殖的影响;采用Western blot观察其对芳香化酶的影响;采用RT-PCR观察其对CYP19基因的影响;采用瞬时转染形成高表达及低表达CYP19基因的MCF-7细胞,并用Western blot的方法观察雷公藤甲素对转染细胞CYP19以及其下游通路相关基因JNK、P38和ERK的影响.结果 雷公藤甲素能明显抑制MCF-7细胞增殖(P<0.01),其抑制作用随着浓度增高和时间的延长而增强,能抑制芳香化酶及CYP19基因的表达,并能明显下调低表达CYP19-MCF-7细胞的CYP19表达,及抑制JNK、p-38和ERK的磷酸化.结论 雷公藤甲素抗MCF-7细胞的作用机制之一,是通过抑制芳香化酶起作用,并引起下游Ras-Raf-MAPK-ERK激酶系统通路受抑制.%OBJECTIVE To investigate the mechanisms of Triptolide in ER(+) human breast cancer MCF-7 cells from CYP19.METHODS The inhibition of the MCF-7 cells influenced by Triptolide and Letrozole was analyzed respectively by MTT assays.The aromatase levels were measured by Western blot.The CYP19 gene was observed by RT-PCR.The MCF-7 cells of high and low expression CYP19 gene were transfacted.It's CYP-19 and downstream channel gene JNK,p-38 and ERK were observed by Western blot.RESULTS Triptolide displayed a dose-and time-dependent inhibition of the MCF-7 cells.It inhibited the aromatase and CYP19.The expression of CYP19 in MCF-7 cells of low expression CYP19 was significantly decreased.Phosphorylation of JNK,p38 and ERK was inhibited too.CONCLUSION Triptolide inhibit MCF-7 cells not only via inhibition aromatase,but also blocking Ras-Raf-MAPK-ERK enzyme.