HDAC抑制剂SAHA对脉络膜黑色素瘤细胞OCM1的作用  

Effect of HDAC inhibitor SAHA on choroidal melanoma OCM1 cells

作　　者：张益萌 杨瀚毅 陈亦玄 宁佳怡 闫小龙 韩静 ZHANG Yimeng;YANG Hanyi;CHEN Yixuan;NING Jiayi;YAN Xiaolong;HAN Jing(Department of Ophthalmology,Second Affiliated Hospital of Air Force Military Medical University,Xi’an 710038,China;Department of Ophthalmology,Xi’an Medical University;Department of Thoracic Surgery,Second Affiliated Hospital of Air Force Military Medical University)

机构地区：[1]空军军医大学第二附属医院眼科,西安710038 [2]西安医学院眼科学教研室 [3]空军军医大学第二附属医院胸外科

出　　处：《山西医科大学学报》2023年第6期733-740,共8页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(8217111370);陕西省科技攻关重点研发计划项目(2021SF-158);唐都医院临床重点研究项目(2021LCYJ019,2018LCYJ008)。

摘　　要：目的探讨组蛋白去乙酰化酶(histone deacetylase,HDAC)抑制剂辛二酰苯胺异羟肟酸(suberoylanilide hydroxamic acid,SAHA)对脉络膜黑色素瘤(choroidal melanoma,CM)细胞系OCM1增殖侵袭的机制。方法使用显微镜观察不同浓度SAHA(0.625,1.25,2.5μmol/L)对OCM1细胞形态结构的影响,利用CCK-8实验观察不同浓度SAHA(0.625,1.25,2.5μmol/L)对OCM1细胞活力的影响,通过EdU染色法和克隆形成实验观察control组(0μmol/L SAHA)和1.25μmol/L SAHA组OCM1细胞增殖的变化,通过划痕实验和迁移侵袭实验观察control组和1.25μmol/L SAHA组OCM1细胞迁移侵袭能力的变化。采用Western blot实验检测不同浓度SAHA(0.625,1.25,2.5μmol/L)组中HDAC4、CyclinD1、c-Myc、E-cadherin和Snail蛋白表达水平。结果与control组相比较,光镜下明显可见0.625μmol/L SAHA组、1.25μmol/L SAHA组、2.5μmol/L SAHA组OCM1细胞增殖能力减弱,促进细胞皱缩和死亡,且细胞增殖活力降低(P<0.05)。与control组比较,1.25μmol/L SAHA组OCM1细胞增殖和迁移侵袭能力明显降低(P<0.05)。此外,与control组相比较,0.625μmol/L SAHA组、1.25μmol/L SAHA组、2.5μmol/L SAHA组E-cadherin蛋白表达逐渐升高,而HDAC4和Snail蛋白表达逐渐降低(P<0.05)。结论SAHA可通过抑制HDAC4明显降低OCM1细胞增殖活力和迁移侵袭能力。Objective To investigate the effect and its mechanism of histone deacetylase(HDAC)inhibitor suberoylanilide hydroxamic acid(SAHA)on the proliferation and invasion abilities of choroidal melanoma(CM)cell line OCM1.Methods The effects of different concentrations of SAHA(0.625,1.25,2.5μmol/L)on the morphological structure of OCM1 cells were observed using a microscope,the effects of different concentrations of SAHA(0.625,1.25,2.5μmol/L)on the viability of OCM1 cells were observed using CCK-8 assay,the proliferation of OCM1 cells in control group(0μmol/L SAHA)and 1.25μmol/L SAHA group was observed by EdU staining and colony formation assay,and the migration and invasion abilities of OCM1 cells in control group and 1.25μmol/L SAHA group were observed by scratch assay and migration and invasion assay.In addition,HDAC4,CyclinD1,c-Myc,E-cadherin and Snail protein expression levels in different concentrations of SAHA groups(0.625,1.25,2.5μmol/L)were detected by Western blot assay.Results Compared with control group,the proliferation ability of OCM1 cells in 0.625μmol/L SAHA group,1.25μmol/L SAHA group and 2.5μmol/L SAHA group was weakened and the cell shrinkage and death was promoted under the light microscope,and the cell proliferation activity was decreased(P<0.05).Compared with control group,the proliferation,migration and invasion abilities of OCM1 cells were significantly decreased in 1.25μmol/L SAHA group(P<0.05).Compared with control group,the E-cadherin protein expression was gradually increased,and the HDAC4 and Snail protein expression was decreased in 0.625μmol/L SAHA group,1.25μmol/L SAHA group,and 2.5μmol/L SAHA group(all P<0.05).Conclusion SAHA can significantly reduce the proliferation viability and the migratory invasion ability of CM cell line OCM1 cells by inhibiting HDAC4.

关 键 词：辛二酰苯胺异羟肟酸(SAHA) 组蛋白去乙酰化酶4 SNAIL 脉络膜黑色素瘤细胞 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R739.7[医药卫生—肿瘤]