摘要
目的研究组蛋白去乙酰化酶抑制剂(HDACis)PCI-24781对上皮性卵巢癌SKOV-3细胞增殖、迁移及Wnt/β-catenin信号通路的影响。方法采用xCELLigence RTCA检测SKOV-3细胞的增殖与迁移,流式细胞仪DCFH-DA探针检测细胞内活性氧(ROS)的生成情况,免疫蛋白印迹法(Western blot)检测Ac-H3、Ac-H4、β-catenin、P53的表达。结果 PCI-24781对上皮性卵巢癌SKOV-3细胞呈剂量-时间依赖性抑制细胞增殖,差异有统计学意义(P<0.05),其48h的半数有效抑制水平(IC50)值为0.193 0μmol/L。PCI-24781对上皮性卵巢癌SKOV-3细胞迁移影响不明显,各组Cell Index差异无统计学意义(P>0.05)。PCI-24781可提高细胞内ROS水平(P<0.05),且高水平组的PCI-24781提高细胞内ROS水平更明显增加。Western blot检测可见随PCI-24781水平升高,β-catenin表达水平减弱、Ac-H3、Ac-H4、P53表达水平增强。结论 PCI-24781可增加细胞内ROS水平,上调组蛋白乙酰化水平,促进抑癌基因P53表达,抑制经典Wnt/β-catenin信号通路,从而抑制细胞增殖,产生抗肿瘤作用。
Objective To study the antitumor effects of Histone deacetylase inhibitors（HDACis）PCI-24781 on the proliferation and migration of epithelial ovarian cancer cell line SKOV-3and the effects on the expression of wnt/β-catenin signaling pathway.Methods xCELLigence RTCA Multi-function real time non labeled cell analyzer was used to detect the proliferation and migration of SKOV-3cells,flow cytometry was used to detect the production of intracellular ROS by DCFH-DA probe,Western blot was used to detect the expression of Ac-H3,Ac-H4andβ-catenin,P53.Results PCI-24781 could inhibit the proliferation of epithelial ovarian cancer cell line SKOV-3in a concentration-time dependent manner,the difference was statistically significant（P〈0.05）,the IC50 value of 48 htreatment was 0.193 0μmol/L.The effect of PCI-24781 on the migration of SKOV-3cells was not significant,and there was no statistical significance（P〉0.05）of Cell Index in each group.PCI-24781 could improve the intracellular ROS level（P〈0.05）,and ROS production increased remarkably in the high concentration PCI-24781 treatment group.Western blot detection showed that with the increase of PCI-24781 concentration,the expression level of β-catenin was decreased,the expression of Ac-H3,Ac-H4 and P53were enhanced.Conclusion PCI-24781 could increase the level of intracellular ROS,increase the level of histone acetylation,up-regulate the expression of tumor suppressor gene P53,inhibit the Wnt/β-catenin signaling pathway,thereby,inhibit cell proliferation,produce antitumor effects.
出处
《检验医学与临床》
CAS
2017年第20期3072-3075,共4页
Laboratory Medicine and Clinic
