Ras-ERK1/2 信号通路通过 HAT1 调控 H4K12ac 促进骨肉瘤的发生发展。
Ras-ERK1/2 signalling promotes the development of osteosarcoma through regulation of H4K12ac through HAT1.
机构信息
a Department of Orthopedics , China-Japan Union Hospital of Jilin University , Changchun , China.
b Department of Ultrasonography , China-Japan Union Hospital of Jilin University , Changchun , China.
出版信息
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):1207-1215. doi: 10.1080/21691401.2019.1593857.
Histone H4 acetylation at lysine 12 (H4K12ac) has been reported to be associated with the poor prognosis of pancreatic cancer. The study intends to study whether H4K12ac participates in regulating the carcinogenic effect of Ras-ERK1/2 on osteosarcoma (OS). The plasmids of pEGFP-N1, pEGFP-Ras and pEGFP-K-Ras were transfected into MG-63 cells, the protein levels of H4K12ac and ERK1/2 were analyzed by Western blot. Effects of H4K12ac on cell proliferation and migration of MG-63 cells were tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2Htetrazolium bromide solution (MTT), transwell, soft-agar colony formation and flow cytometry assays. Effect of H4K12ac on the transcription of ERK1/2 downstream genes was analyzed by RT-qPCR and ChIP assays. The involvements of HDAT6, HAT1 and MDM2 in cell proliferation and migration of MG-63 cells were finally studied. We found that H4K12ac was specifically down-regulated by Ras-ERK1/2 activation in MG-63 cells. H4K12ac suppressed Ras-ERK1/2-induced cell viability, colony formation and migration in MG-63 cells. Additionally, HDAC6 silence recovered Ras-ERK1/2-repressed H4K12ac expression, as well as inhibited the carcinogenic effect of Ras-ERK1/2 on MG-63 cells. Besides, down-regulated H4K12ac induced by Ras-ERK1/2 was found to be associated with MDM2-mediated HAT1 degradation. In conclusion, these results testified that Ras-ERK1/2 signalling promoted the development of OS by mediating H4K12ac through MDM2-mediated HAT1 degradation.
组蛋白 H4 赖氨酸 12 乙酰化（H4K12ac）已被报道与胰腺癌的不良预后相关。本研究旨在研究 H4K12ac 是否参与调节 Ras-ERK1/2 对骨肉瘤（OS）的致癌作用。将 pEGFP-N1、pEGFP-Ras 和 pEGFP-K-Ras 质粒转染到 MG-63 细胞中，通过 Western blot 分析 H4K12ac 和 ERK1/2 的蛋白水平。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H 四唑溴盐溶液（MTT）、Transwell、软琼脂集落形成和流式细胞术检测 H4K12ac 对 MG-63 细胞增殖和迁移的影响。通过 RT-qPCR 和 ChIP 分析检测 H4K12ac 对 ERK1/2 下游基因转录的影响。最后研究了 HDAT6、HAT1 和 MDM2 在 MG-63 细胞增殖和迁移中的作用。结果发现，在 MG-63 细胞中，H4K12ac 被 Ras-ERK1/2 激活特异性下调。H4K12ac 抑制 Ras-ERK1/2 诱导的 MG-63 细胞活力、集落形成和迁移。此外，沉默 HDAC6 恢复了 Ras-ERK1/2 抑制的 H4K12ac 表达，并抑制了 Ras-ERK1/2 对 MG-63 细胞的致癌作用。此外，发现 Ras-ERK1/2 诱导的 H4K12ac 下调与 MDM2 介导的 HAT1 降解有关。综上所述，这些结果表明，Ras-ERK1/2 信号通过 MDM2 介导的 HAT1 降解来调节 H4K12ac，从而促进 OS 的发展。
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