SGI-1776钝化Pim-1对卵巢癌细胞增殖和侵袭的抑制作用及其机制(英文)

SGI-1776, an imidazo pyridazine compound, inhibits the proliferation of ovarian cancer cells by inactivating Pim-1

目的:检测SGI-1776对卵巢癌HO-8910细胞增殖、迁移、侵袭的抑制作用及其可能机制。方法:体外培养卵巢癌HO-8910细胞;MTT法和克隆形成法检测SGI-1776对HO-8910细胞的增殖抑制作用;PI染色流式细胞术(flow cytometry,FCM)检测SGI-1776对HO-8910细胞周期分布的影响;Transwell法检测SGI-1776对HO-8910细胞迁移及侵袭的抑制作用;ELISA,Western印迹,siRNA/cDNA转染检测SGI-1776对HO-8910细胞增殖、迁移、侵袭抑制作用的可能分子机制。结果:SGI-1776在体外对HO-8910细胞增殖、迁移、侵袭呈现出浓度依赖性的抑制作用,阻滞细胞周期于G1期。随Pim-1激酶活性降低,Pim-1蛋白表达下调,同时Pim-1下游蛋白CDK6,pCDK6,CDK4,pCDK4,CDK2和pCDK2表达下调,而P21和P27蛋白表达上调。用siRNA干扰沉默Pim-1基因,则SGI-1776对HO-8910细胞增殖、迁移、侵袭的抑制作用明显加强;用Pim-1-cDNA转染HO-8910细胞,则能部分抵消SGI-1776对细胞增殖、迁移、侵袭的抑制作用。结论:SGI-1776通过钝化Pim-1而发挥其抑制卵巢癌HO-8910细胞增殖、迁移和侵袭的作用,提示Pim-1是卵巢癌治疗的新的分子靶。

Objective: To investigate the antitumor effect of SGI-1776 on human ovarian cancer HO-8910 cells and its molecular mechanism. Methods: HO-8910 cells were cultured in vitro, and the proliferation inhibitory effects of SGI-1776 were determined by MTT assay and colony formation assay. The effect of SGI-1776 on the distribution of cell cycle phase was observed by 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 ELISA, Western blot, siRNA and cDNA transfection were used to explore the molecular mechanism.Results: SGI-1776 presented dramatic anti-tumor activity against HO-8910 cells in vitro, inhibited the cells proliferation and colony formation, and attenuated the migration and invasion in a dose-dependent manner, accompanied by cell cycle arrest in G1 phase. SGI-1776 caused the proliferation inhibition with concomitant decrease in Pim-1 kinase activity, down-regulated the expression of Pim-1 protein and and its downstream genes, such as CDK6, pCDK6, CDK4, pCDK4, CDK2 and pCDK2, and increased the expression of P21 and P27. Down-regulation expression of Pim-1 by siRNA followed SGI-1776 treatment resulted in enhanced cell proliferation inhibition rate and attenuated migration/invasion. Up-regulation of Pim-1 by cDNA transfection attenuated SGI-1776-induced cell proliferation inhibition and its migration/invasion.Conclusion: Pim-1 mediates the biological effect of SGI-1776 in human ovarian cancer HO-8910 cells, suggesting Pim-1 might be a novel target for human ovarian cancer.