Colon cancer is a prevalent clinical malignant tumor of the digestive system. The current study aims to explore the miR-144 expression in colorectal cancer (CRC) cell lines and CRC stem cells (CSCs) and to explore its effect on the stemness of CSCs and the targeted regulation of Krüppel-like factor 4 (KLF4). Use qRT-PCR to detect the expression level of miR-144 in CRC cells SW480, HCT116, and H129 and the healthy colon cell NCM460. The CSCs that were used were cultured in HCT116 cells. Use western blot to explore the expressions of Nanog, SOX2, and OCT4 stemness marker protein. After it was transfected with miR-144 mimics or KLF4 plasmid, use MTT to explore the cell viability of CSCs, use flow cytometry to evaluate apoptosis, and use transwell assay to evaluate the ability of invasive of CSCs. The targeting effect of miR-144 on the KLF4 gene was verified using TargetScan prediction and the double-luciferase reporter gene test. Use qRT-PCR to evaluate the role of miR-144 mimics on KLF4 mRNA expression in CSCs. The qRT-PCR results exhibited that the miR-144 expression in CRC cells was higher than that in the healthy colon cell line. The expressions of OCT4, Nanog, and SOX2 stem cell markers were up-regulated in CSCs, and the expression of miR144 increased in CSCs. The cell viability, apoptosis, and invasion of CSCs increased after miR-144 was transfected. The TargetScan prediction and double-luciferase reporter gene assay confirmed that miR-144 was targeted by KLF4, and the expression of KLF4 mRNA in the miR-144 mimics group reduced. Moreover, the overexpression of KLF4 could partially reverse the role of miR-144 mimics on CSCs. In summary, miR-144 was highly expressed in CRC cell lines and CSCs, and the overexpression of miR-144 in CSCs significantly promoted the proliferation of CSCs, inhibited its apoptosis, and promoted its invasion ability. In addition, its preliminary mechanism, possibly through negative regulation KLF4, promotes the stemness of CSCs, and miR-144 is likely to be a potential target for eliminating CSC from CRC treatment.

中文翻译：

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靶向Krüppel样因子4抑制miR-144抑制结肠癌干细胞干性的初步研究。

结肠癌是消化系统流行的临床恶性肿瘤。当前的研究旨在探讨miR-144在结直肠癌（CRC）细胞系和CRC干细胞（CSCs）中的表达，并探讨其对CSCs干性的影响以及Krüppel样因子4（KLF4）的靶向调控。使用qRT-PCR检测miR-144在CRC细胞SW480，HCT116和H129和健康结肠细胞NCM460中的表达水平。使用的CSC在HCT116细胞中培养。使用蛋白质印迹来研究Nanog，SOX2和OCT4干性标记蛋白的表达。用miR-144模拟物或KLF4质粒转染后，使用MTT探索CSCs的细胞生存力，使用流式细胞仪评估细胞凋亡，并使用Transwell分析法评估CSCs的侵袭能力。使用TargetScan预测和双荧光素酶报告基因测试验证了miR-144对KLF4基因的靶向作用。使用qRT-PCR评估miR-144模拟物对CSC中KLF4 mRNA表达的作用。qRT-PCR结果显示，CRC细胞中的miR-144表达高于健康结肠细胞系。CSCs中OCT4，Nanog和SOX2干细胞标志物的表达上调，而miR144的表达在CSCs中增加。miR-144转染后，细胞活力，凋亡和CSC侵袭增加。TargetScan预测和双荧光素酶报告基因检测证实，miR-144被KLF4靶向，miR-144模仿组中KLF4 mRNA的表达降低。此外，KLF4的过表达可以部分逆转miR-144模拟物在CSC上的作用。综上所述，miR-144在CRC细胞系和CSCs中高表达，miR-144在CSCs中的过表达显着促进CSCs的增殖，抑制其凋亡并增强其侵袭能力。此外，其初步机制可能通过负调控KLF4促进CSC的干性，而miR-144可能是从CRC治疗中消除CSC的潜在目标。