7-二氟甲氧基-5,4'-二正辛烷氧基金雀异黄素钝化Pim-1对宫颈癌细胞增殖和侵袭的抑制作用

目的 检测7-二氟甲氧基-5,4''-二正辛烷氧基金雀异素(7-difluoromethoxyl-5,4''-di-n-octylygenistein，DFOG)对宫颈癌HeLa细胞增殖、迁移、侵袭的抑制作用及其机制。方法 体外培养宫颈癌HeLa细胞，用MTT法检测DFOG对HeLa细胞的增殖抑制作用；用PI染色FCM检测DFOG对HeLa细胞周期分布的影响；用Transwell法检测DFOG对HeLa细胞迁移及侵袭的抑制作用；用Western blotting、siRNA/cDNA转染检测DFOG对HeLa细胞增殖、迁移侵袭抑制作用的可能分子生物学机制。结果 DFOG在体外对HeLa细胞增殖、迁移、侵袭呈浓度依赖性的抑制作用，阻滞细胞周期于G1期，伴随着Pim-1蛋白表达下调，同时Pim-1下游蛋白Cyclin D、Cyclin E、Cyclin A表达下调，而p15和p21蛋白表达上调。用siRNA干扰沉默Pim-1基因，则DFOG对HeLa细胞增殖、迁移、侵袭的抑制作用明显加强；用Pim-1-cDNA转染HeLa细胞，则能部分抵消DFOG对细胞增殖、迁移、侵袭的抑制作用。结论 DFOG通过钝化Pim-1组织细胞与G1期而发挥其抑制宫颈癌HeLa细胞增殖、迁移和侵袭的作用。

Effects of 7-difluoromethoxyl-5,4'-di-n-octylygenistein on the Actions of Inhibiting Proliferation and Invasion of Cervical Cancer Cells via Inactivating Pim-1

OBJECTIVE To investigate the anti-tumor effects of 7-difluoromethoxyl-5,4''-di-n-octylygenistein (DFOG) on human cervical cancer HeLa cells and its possible molecular mechanism. METHODS HeLa cells were cultured in vitro, and the proliferation inhibitory effects of DFOG was determined using MTT assay. The effect of DFOG on distribution of cell cycle phase was observed using flow cytometry with propidium iodide (PI) staining. The inhibition rate of migration and invasion were valued by Transwell cell assay. Multiple molecular techniques, such as Western blotting, siRNA and cDNA transfection were used to explore the molecular mechanism. RESULTS DFOG presented dramatic anti-tumor activity against HeLa cells in vitro, inhibited the cells proliferation, attenuated the migration and invasion ability in dose-dependent manner, accompanied by cell cycle arrest in G1 phase. DFOG caused proliferation inhibition accompanied with the expression down-regulation of Pim-1 protein and its downstream genes, such as Cyclin D, Cyclin E, Cyclin A, and increased expression p15 and p21. Down-regulation expression of Pim-1 by siRNA followed DFOG treatment resulted in cell proliferation inhibition rate enhanced and migration/invasion ability attenuated. Up-regulation expression of Pim-1 by cDNA transfection attenuated DFOG-induced cell proliferation inhibition and its migration/invasion capability. CONCLUSION DFOG can inhibit proliferation and invasion of human cervical cancer HeLa cells in vitro through inactivated Pim-1 expression accompanied cells in G1 phase.