■结直肠癌(CRC)是一种常见的消化道恶性肿瘤。它的特点是恶性程度高,早期转移和预后不良。研讨显示miR-369-3p对多种肿瘤的生物学功效具有必定的感化。然而,miR-369-3p及其潜在靶基因参与CRC发病的机制尚未阐明.本研究旨在研究miR-369-3p与转录因子4(TCF4)的关系,揭示CRC发生发展的机制,并为CRC的治疗提供有希望的靶点。
■使用实时定量聚合酶链反应(RT-qPCR)检测CRC组织和细胞中的miR-369-3p水平。将miR-369-3p模拟物和/或TCF4过表达载体转染到SW480细胞中。RT-qPCR检测miR-369-3p和TCF4mRNA的表达。生物信息学分析预测miR-369-3p与TCF43'UTR的结合位点,并通过双荧光素酶报告基因检测验证了靶向关系。使用BrdU抗体和Transwell测定法通过标记免疫荧光测定法研究细胞增殖和侵袭。通过商业试剂盒测定细胞的氧化应激能力。通过蛋白质印迹法测量与细胞增殖和侵袭相关的蛋白质的水平。
■miR-369-3p水平在CRC组织和细胞系中显著下调,尤其是SW480细胞(P<0.05)。TCF4与miR-369-3p的表达呈负相关。高水平的miR-369-3p靶向TCF4抑制细胞增殖并下调Ki67和PCNA的蛋白表达(P<0.05)。过表达miR-369-3p结合TCF4抑制细胞侵袭,降低血管内皮生长因子(VEGF)和E-cadherin蛋白水平(P<0.05)。此外,miR-369-3p的上调可阻断TCF4的表达,提高超氧化物歧化酶(SOD)活性,降低丙二醛(MDA)含量和乳酸脱氢酶(LDH)活性(P<0.05)。
■MiR-369-3p抑制增殖,通过靶向TCF4对CRC细胞的侵袭和氧化应激,从而将miR-369-3p定义为诊断和治疗CRC的潜在靶标。
UNASSIGNED: Colorectal carcinoma (CRC) is a common malignant tumor of the digestive tract. It is characterized by a high degree of malignancy, early metastasis and poor prognosis. Studies have shown the effect of miR-369-3p on the biological function of a variety of tumors. However, the mechanism by which miR-369-3p and its potential target genes participate in the pathogenesis of CRC has not been elucidated. This
study aims to
study the relationship between miR-369-3p and transcription factor 4 (TCF4), to reveal the mechanism of the occurrence and development of CRC, and to provide a promising target for the treatment of CRC.
UNASSIGNED: Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the miR-369-3p levels in CRC tissues and cells. miR-369-3p mimics and/or TCF4 overexpression vectors were transfected into SW480 cells. The expression of miR-369-3p and TCF4 mRNA was detected using RT-qPCR. Bioinformatics analysis predicted the binding site of miR-369-3p to the TCF4 3\'UTR, and the targeting relationship was verified by a dual luciferase reporter gene assay. Cell proliferation and invasion were investigated by labeled immunofluorescence assay using BrdU antibody and Transwell assay. The oxidative stress ability of cells was determined by commercial kits. The levels of proteins related to cell proliferation and invasion were measured by western blotting.
UNASSIGNED: The level of miR-369-3p was significantly down-regulated in CRC tissues and cell lines, especially in SW480 cells (P<0.05). The expression of TCF4 was negatively correlated with that of miR-369-3p. High levels of miR-369-3p targeting TCF4 suppressed cell proliferation and downregulated the protein expression of Ki67 and PCNA (P<0.05). Overexpressed miR-369-3p binding TCF4 inhibited cell invasion and decreased the protein levels of vascular endothelial growth factor (VEGF) and E-cadherin (P<0.05). Furthermore, upregulation of miR-369-3p increased the activity of superoxide dismutase (SOD) while decreasing the content of malondialdehyde (MDA) and activity of lactate dehydrogenase (LDH) by blocking the expression of TCF4 (P<0.05).
UNASSIGNED: MiR-369-3p inhibits the proliferation, invasion and oxidative stress of CRC cells by targeting TCF4, thus defining miR-369-3p as a potential target for the diagnosis and treatment of CRC.