多种癌症的肿瘤发生受到表观遗传失调的影响。在检查了591个表观遗传调节因子（ERFs）后，我们发现AF9在结直肠癌（CRC）中能够显著抑制恶性肿瘤的扩散，这是基于我们的创伤愈合分析而得出的结果。然而，AF9在CRC中的确切作用尚待探索。 为了研究AF9在CRC中的功能，我们使用小干扰RNA（siRNA）来沉默591个ERFs的表达。随后，我们进行创伤愈合实验来评估细胞增殖和迁移情况。我们进行了体外和体内实验来阐明AF9对CRC的潜在影响。我们分析了临床样本以评估AF9表达与CRC预后的关联。此外，我们采用Azoxymethane-Dextran Sodium Sulfate（AOM/DSS）诱导的CRC AF9IEC-/-小鼠模型来确认AF9在CRC中的作用。为了确定AF9的靶基因，我们进行了RNA测序和共免疫沉淀分析。此外，我们运用生物信息学预测法来识别可能靶向AF9的miRNAs。 在检查了591个ERFs后，我们发现AF9在CRC中表达下调，并且与CRC患者的长期生存率呈正相关。体内和体外实验证明，AF9的缺失促进了细胞增殖、迁移以及糖酵解。具体而言，将MLLT3（AF9）基因敲除在肠道上皮细胞中，显著增加了AOM/DSS诱导的肿瘤形成。我们还发现miR-145可以靶向AF9的3'非翻译区域抑制AF9的表达。AF9的缺失导致糖异生基因的表达下调，包括磷酸烯醇式丙酮酸羧激酶2（PCK2）和果糖1,6-二磷酸酶1（FBP1），随后促进葡萄糖消耗和肿瘤发生。 AF9对PCK2和FBP1的上调是必不可少的，miR-145/AF9轴的破坏可能成为CRC治疗发展的潜在靶点。© 2023 The Authors. Clinical and Translational Medicine由上海临床生物信息研究所代表John Wiley & Sons Australia, Ltd出版。

The tumourigenesis of various cancers is influenced by epigenetic deregulation. Among 591 epigenetic regulator factors (ERFs) examined, AF9 showed significant inhibition of malignancy in colorectal cancer (CRC) based on our wound healing assays. However, the precise role of AF9 in CRC remains to be explored.To investigate the function of AF9 in CRC, we utilised small interfering RNAs (siRNAs) to knock down the expression of 591 ERFs. Subsequently, we performed wound healing assays to evaluate cell proliferation and migration. In vitro and in vivo assays were conducted to elucidate the potential impact of AF9 in CRC. Clinical samples were analysed to assess the association between AF9 expression and CRC prognosis. Additionally, an Azoxymethane-Dextran Sodium Sulfate (AOM/DSS) induced CRC AF9IEC-/- mouse model was employed to confirm the role of AF9 in CRC. To identify the target gene of AF9, RNA-seq and coimmunoprecipitation analyses were performed. Furthermore, bioinformatics prediction was applied to identify potential miRNAs that target AF9.Among the 591 ERFs examined, AF9 exhibited downregulation in CRC and showed a positive correlation with prolonged survival in CRC patients. In vitro and in vivo assays proved that depletion of AF9 could promote cell proliferation, migration as well as glycolysis. Specifically, knockout of MLLT3 (AF9) in intestinal epithelial cells significantly increased tumour formation induced by AOM/DSS. We also identified miR-145 could target 3'untranslated region of AF9 to suppress AF9 expression. Loss of AF9 led to decreased expression of gluconeogenic genes, including phosphoenolpyruvate carboxykinase 2 (PCK2) and fructose 1,6-bisphosphatase 1 (FBP1), subsequently promoting glucose consumption and tumourigenesis.AF9 is essential for the upregulation of PCK2 and FBP1, and the disruption of the miR-145/AF9 axis may serve as a potential target for the development of CRC therapeutics.© 2023 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.