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
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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)


Effects and related mechanisms of histone deacetylase inhibitors on the proliferation of choroidal melanoma C918 cell line
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National Natural Science Foundation of China(No.8217111370); Key Research and Development Program of Shaanxi Province for Science and Technology Research(No.2021SF-158); Key Clinical Research Project of Tangdu Hospital(No.2021LCYJ019, 2018LCYJ008)

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    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。

    Abstract:

    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 2.5 μmol/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 2.5 μmol/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, 1.25 μmol/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.

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张益萌,杨瀚毅,宁佳怡,等. 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.国际眼科杂志, 2023,23(2):193-197.

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  • 收稿日期:2022-06-06
  • 最后修改日期:2023-01-16
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  • 在线发布日期: 2023-02-02
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