SALL4 是一种在多种癌症中高表达的转录因子，并被认为参与了癌症的发展。在结肠癌（CC）中，SALL4 被认为具有促癌作用，但其分子机制尚不清楚。我们进行了染色质免疫沉淀实验和双荧光素酶分析实验以验证 SALL4 与 ROBO2 的结合关系。利用 qRT-PCR 检测了 SALL4 和 ROBO2 的 mRNA 表达水平，利用流式细胞术分析了细胞周期分布。利用免疫印迹检测了 SALL4 的表达以及与细胞周期/干细胞相关的蛋白质。通过 MTT、克隆形成和球形形成实验阐明了 SALL4 和 ROBO2 对细胞增殖能力和肿瘤细胞干性的影响。结果显示，SALL4 和 ROBO2 在 CC 中上调表达，并且 SALL4 可以激活 ROBO2 的转录。下调 SALL4 能够显著抑制 CC 细胞的增殖能力，并通过抑制 cyclin B、cyclin E 和 cyclin D1 的表达来阻滞细胞周期在 G0/G1 阶段。此外，救治实验结果表明，上调 ROBO2 能够扭转下调 SALL4 对 CC 细胞增殖的抑制作用，加速细胞周期的进展，从而促进肿瘤干细胞的球形形成。SALL4 通过直接激活 ROBO2 的表达来推进 CC 的增殖和细胞干性，并揭示了 SALL4 在 CC 中的新机制，并指出 SALL4/ROBO2 轴很可能是 CC 临床治疗的潜在靶点。

SALL4 is a transcription factor highly expressed in diverse cancers and is implicated in the development of cancer. SALL4 has been implied to play a cancer-promoting role in colon cancer (CC), but the molecular mechanism remains unclear. Chromatin immunoprecipitation assay and dual-luciferase assay were conducted to verify the binding relationship of SALL4 and ROBO2. qRT-PCR detected the mRNA expression levels of SALL4 and ROBO2, and the flow cytometry analyzed the cell cycle distribution. Western blot examined SALL4 expression, and cell cycle/cell stemness-related proteins. The impact of SALL4 and ROBO2 on the proliferation capacity of cells and tumor cell stemness was elucidated by MTT, colony formation, and sphere-forming assays. SALL4 and ROBO2 were up-regulated in CC, and SALL4 could activate the transcription of ROBO2. Down-regulated SALL4 was able to significantly restrain the proliferation capacity of CC cells and arrest the cell cycle in G0/G1 phase by repressing the expression of cyclin B, cyclin E, and cyclin D1. Besides, the rescue assay results indicated that up-regulated ROBO2 could reverse the repressive impact of down-regulated SALL4 on the proliferation of CC cells and accelerate the progression of the cell cycle, thus promoting the sphere-forming of tumor stem cells. SALL4 advanced the proliferation of CC and cell stemness through direct activation of ROBO2 expression, implied the novel mechanism of SALL4 in CC, and pointed out that SALL4/ROBO2 axis was likely to be a potential target for clinical treatment of CC.