Department of General Surgery, Juancheng People's Hospital, Heze, China.
Department of Gastroenterology, Heze Municipal Hospital, Heze, China.
J Cell Physiol. 2020 Apr;235(4):3905-3915. doi: 10.1002/jcp.29284. Epub 2019 Oct 22.
H3 modification is related to a wide range of tumors, including liver cancer. The Ras passageway is actuated in human diseases. Thus, we investigated the roles of Ras in liver cancer cells via acetylation of H3K56. Ras-carrying G12V and Y40C site mutation was transfected into liver cancer cell lines SNU-475 and SK-Hep-1. Acetylation of H3K56 and phosphatidylinositol 3-kinase (PI3K), P300/CBP-associated factor (PCAF) and Mouse double minute 2 homolog (MDM2) was tested via western blot. Cell activity, colonies, and migration were tested via Cell Counting Kit-8, soft-agar colony formation, and Transwell experiment, respectively. Sirtuin 6 (SIRT6) and PCAF were tested via quantitative reverse transcription polymerase chain reaction (qRT-PCR). Chromatin immunoprecipitation was employed to test the relationship between Ras and downstream elements. Flow cytometry was employed to test cell cycle series. We found that Ras transfection reduced the acetylation of H3K56 and activated phosphorylation of protein kinase B. H3K56Q (H3K56ac overexpression) suppressed cell activity, colonies, and migration. H3K56ac changed Ras downstream factors expression. Ras bound to Ras-PI3K downstream elements' promoters. SIRT6 silencing raised H3K56ac and suppressed cell activity, migration and S phage cell percentage. SIRT6 silence transformed expression of downstream elements. PCAF and H3K56ac demonstrated the close current while MDM2 was conversed. In summary, the Ras-PI3K passageway promoted cell growth and metastasis via decreasing H3K56ac, in which MDM2-mediated PCAF was involved.
H3 修饰与广泛的肿瘤有关，包括肝癌。 Ras 通路在人类疾病中被激活。因此，我们通过 H3K56 的乙酰化研究了 Ras 在肝癌细胞中的作用。将携带 Ras 的 G12V 和 Y40C 位点突变的质粒转染到肝癌细胞系 SNU-475 和 SK-Hep-1 中。通过 Western blot 检测 H3K56 乙酰化和磷脂酰肌醇 3-激酶（PI3K）、P300/CBP 相关因子（PCAF）和鼠双微体 2 同源物（MDM2）。通过 Cell Counting Kit-8 检测细胞活性、集落形成、Transwell 实验检测细胞迁移。通过定量逆转录聚合酶链反应（qRT-PCR）检测 Sirtuin 6（SIRT6）和 PCAF。通过染色质免疫沉淀检测 Ras 与下游元件的关系。通过流式细胞术检测细胞周期系列。我们发现 Ras 转染降低了 H3K56 的乙酰化并激活了蛋白激酶 B 的磷酸化。H3K56Q（H3K56ac 过表达）抑制细胞活性、集落形成和迁移。H3K56ac 改变了 Ras 下游因子的表达。Ras 结合 Ras-PI3K 下游元件的启动子。沉默 Sirtuin 6 提高了 H3K56ac 并抑制了细胞活性、迁移和 S 期细胞百分比。Sirtuin 6 沉默改变了下游元件的表达。PCAF 和 H3K56ac 表现出密切的关系，而 MDM2 则相反。总之，Ras-PI3K 通路通过降低 H3K56ac 促进细胞生长和转移，其中涉及 MDM2 介导的 PCAF。
