目的 揭示HCT116细胞中miR-513b-5p通过IL-6/STAT3抑制结肠癌干细胞（CCSCs）转移的机制。采用球形形成培养基和磁性细胞分选技术对CCSCs进行富集和筛选。我们使用集落形成测定、细胞增殖和活力测定以及裸鼠移植肿瘤测定来鉴定 CCSC。通过ELISA鉴定细胞培养基中的IL-6，并比较不同细胞群的生长、活力、伤口愈合和跨孔迁移，以区分它们。进行双荧光素酶报告基因检测、RT-PCR和/或Western Blot分析以确定它们之间的相关性。CD133 CD44 HCT116细胞被证明具有更高的克隆效率、更强的增殖能力和活力以及更强的致瘤性。双荧光素酶报告基因检测显示 miR-513b-5p 对 STAT3 表达产生负面影响。 RT-PCR 和/或 Western Blot 分析表明 miR-513b-5p 对 STAT3 和 Vimentin 产生负面影响，同时对 E-cadherin 表达产生正面影响。实验中STAT3过表达载体  miR-513b-5p抑制剂细胞组效率最高、增殖能力和活力最强、IL-6水平最高。Mir-513b-5p抑制CCSCs的上皮间质转化（EMT）通过 IL-6/STAT3。这种潜在机制可能为结肠癌提供新的治疗靶点。© 2024。作者。

To reveal the mechanisms by which miR-513b-5p inhibits metastasis of colon cancer stem cells (CCSCs) through IL-6/STAT3 in HCT116 cells.Sphere formation media and magnetic cell sorting were used to enrich and screen CCSCs. We used a colony formation assay, cell proliferation and viability assays, and a nude mouse transplantation tumor assay to identify CCSCs. ELISA was performed to identify IL-6 in the cell culture medium, and the growth, viability, wound healing, and transwell migration of distinct cell groups were compared to differentiate them. Dual-luciferase reporter assay, RT-PCR, and/or Western Blot analysis were conducted to determine the correlation between them.CD133+CD44+ HCT116 cells were shown to have higher cloning efficiency, greater proliferation ability and viability, and stronger tumorigenicity. A dual-luciferase reporter assay revealed that miR-513b-5p negatively affected STAT3 expression. RT-PCR and/or Western Blot analysis suggested that miR-513b-5p negatively affected STAT3 and Vimentin, while positively affecting E-cadherin expression. The STAT3 overexpression vector + miR-513b-5p inhibitor cell group had the highest efficiency, greatest proliferation ability and viability, and the highest IL-6 level in the experiments.Mir-513b-5p inhibited the epithelial-mesenchymal transition (EMT) of CCSCs through IL-6/STAT3. This potential mechanism may provide a new therapeutic target for colon cancer.© 2024. The Author(s).