a Department of Gastrointestinal Surgery, Jining No. 1 People's Hospital , Jining , China.
b Affiliated Jining No.1 People's Hospital of Jining Medical University, Jining Medical University , Jining , China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):2754-2763. doi: 10.1080/21691401.2019.1617725.
Colon cancer is one of the most malignant cancers. Histone modification is closely related to tumour development. Our study explored the functions of anti-silencing function 1A (ASF1A) on H4 in colon cancer cells. Colon cancer cell lines and clinical specimens were obtained and/or transfected with full length ASF1A or interference mRNA to mimic or silence of ASF1A expression. Immunoprecipitation and GST pull down was used to target targeting ASF1A or H4. Cells were transfected with H4- or H4-expressing. An kinase activity assay was set to determine whether ASF1A could phosphorylate H4. The severity of autophagy was measured by detecting number of autophagosomes, number of EGFP-LC3, LC3-II/I, percentage of degradation and expression of autophagy associated gene (ATG). ASF1A positively regulated H4; Immunoprecipitation assay and GST pull down results showed that ASF1A interacted directly with H4. In addition, ASF1A silence inhibited autophagosomes number, EGFP-LC3 number, LC3-II/I, percentage of degradation and ATG expression. Moreover, H4 impaired the promoting autophagy effects of ASF1A. The ASF1A-H4 axis promoted colon cancer autophagy via transcriptional regulation of ATG genes. ASF1A regulated H4 and promotes autophagy in colon cancer cells via a kinase activity through regulation of ATG.
结肠癌是最恶性的癌症之一。组蛋白修饰与肿瘤的发生密切相关。我们的研究探讨了抗沉默功能 1A（ASF1A）对结肠癌细胞中 H4 的作用。获取并/或转染全长 ASF1A 或干扰 mRNA 以模拟或沉默 ASF1A 表达，以获得结肠癌细胞系和临床标本。用 GST 下拉和免疫沉淀来靶向 ASF1A 或 H4。用 H4 或 H4 转染细胞。设置激酶活性测定以确定 ASF1A 是否可以磷酸化 H4。通过检测自噬体的数量、EGFP-LC3 的数量、LC3-II/I 的比例、降解的百分比和自噬相关基因（ATG）的表达来测量自噬的严重程度。ASF1A 正向调节 H4；免疫沉淀和 GST 下拉结果表明 ASF1A 与 H4 直接相互作用。此外，ASF1A 沉默抑制自噬体数量、EGFP-LC3 数量、LC3-II/I、降解百分比和 ATG 表达。此外，H4 削弱了 ASF1A 促进自噬的作用。ASF1A-H4 轴通过转录调控 ATG 基因促进结肠癌自噬。ASF1A 通过调节 ATG 来调节 H4 并通过激酶活性促进结肠癌细胞自噬。
