HDAC抑制剂对脉络膜黑色素瘤细胞系C918细胞增殖的影响及相关机制  

Effects and related mechanisms of histone deacetylase inhibitors on the proliferation of choroidal melanoma C918 cell line

作　　者：张益萌 杨瀚毅 宁佳怡 闫小龙 韩静 Yi-Meng Zhang;Han-Yi Yang;Jia-Yi Ning;Xiao-Long Yan;Jing Han(Xi'an Medical University,Xi an 710068,Shaanxi Province,China;Department of Ophthalmology,the Second Affiliated Hospital of the Air Force Military Medical University,Xi'an 710038,Shaanxi Province,China;Department ofThoracic Surgery,the Second Affiliated Hospital of the Air Force Military Medical University,Xi'an 710038,Shaanxi Province,China)

机构地区：[1]西安医学院,陕西省西安市710068 [2]空军军医大学第二附属医院眼科,陕西省西安市710038 [3]空军军医大学第二附属医院胸外科,陕西省西安市710038

出　　处：《国际眼科杂志》2023年第2期193-197,共5页International Eye Science

基　　金：国家自然科学基金资助项目(No.8217111370);陕西省科技攻关重点研发计划项目(No.2021SF-158);唐都医院临床重点研究项目(No.2021LCYJ019,2018LCYJ008)。

摘　　要：目的:阐明组蛋白去乙酰化酶(HDAC)抑制剂辛二酰苯胺异羟肟酸(SAHA)对脉络膜黑色素瘤(CM)细胞系C918细胞增殖的影响并探讨相关机制。方法:使用倒置荧光显微镜观察不同浓度SAHA(0.625、1.25、2.5μmol/L)对C918细胞形态的影响;CCK-8法观察C918细胞活力的变化;细胞平板克隆形成实验和EdU染色法观察SAHA对C918细胞增殖的影响;同时,Western blot检测细胞增殖相关蛋白c-Myc、细胞周期蛋白CyclinA2和CDK2以及HDAC7和成纤维细胞生长因子18(FGF18)的表达。结果:与空白对照组比较,光镜下见SAHA可减小C918的细胞密度,促进细胞皱缩,且随着SAHA浓度的增加对细胞的抑制作用也增强;CCK-8法检测结果显示SAHA浓度依赖性抑制C918细胞活力,2.5μmol/L浓度时抑制率达80%;Western blot结果表明SAHA可呈浓度依赖地降低C918细胞中的增殖蛋白c-Myc、细胞周期蛋白CyclinA2和CDK2的表达;另外,1.25μmol/L SAHA显著降低EdU染色阳性细胞数和细胞克隆数。更为重要的是,与空白对照组相比,SAHA能浓度依赖地降低HDAC7和FGF18的表达。结论:SAHA能够通过抑制HDAC7/FGF18信号通路抑制CM细胞系C918细胞的增殖。AIM:To elucidate the effect of histone deacetylase(HDAC)inhibitor suberoylanilide hydroxamic acid(SAHA)on the proliferation of choroidal melanoma(CM)cell line C918 and to explore the related mechanism.METHODS:Inverted fluorescence microscope was used to observe the effect of different concentrations of SAHA(0.625,1.25 or 2.5μmol/L)on the morphology of C918 cell.The cell viability was detected by cholecystokinin octapeptide(CCK-8)assay.Plate clone formation assay and EdU staining were carried out to measure the effect of SAHA on the cell proliferation.Meanwhile,the expressions of cell proliferation-related proteins including c-Myc,CyclinA2 and CDK2,and histone deacetylase 7(HDAC7)and fibroblast growth factor 18(FGF18)were detected by Western blot.RESULTS:Compared with the control group,the cell density was reduced in SAHA.SAHA could also promote cell shrinkage,and the inhibition on cell was in a concentration-dependent manner.CCK-8 assay showed that SAHA treatment decreased cell viability in a dose-dependent manner and the inhibition rate was 80%when SAHA at 2.5μmol/L.Compared with the control group,Western blot showed that SAHA could suppress the expression of cell proliferation proteins including c-Myc,CyclinA2 and CDK2 in a dose-dependent manner.In addition,1.25μmol/L SAHA significantly decreased the numbers of EdU staining positive cells and cell clones.More importantly,SAHA could dose-dependently decrease the expression of HDAC7 and FGF18 compared with control group.CONCLUSION:SAHA could inhibit the proliferation of CM cell line C918 by inhibiting the HDAC7/FGF18 signaling pathway.

关 键 词：辛二酰苯胺异羟肟酸 组蛋白去乙酰化酶(HDAC) 成纤维细胞生长因子18 细胞增殖 细胞周期 

分 类 号：R739.7[医药卫生—肿瘤]