Department of General Surgery, Chongming Branch of Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, China (mainland).
Med Sci Monit. 2021 Mar 16;27:e929774. doi: 10.12659/MSM.929774.
BACKGROUND MicroRNAs (miRNAs) are novel biomarkers that are important in tumorigenesis and cancer treatment resistance. miR-451 is expressed in human papillary thyroid carcinoma (PTC) tissues and is associated with tumor progression. This study investigated the molecular mechanism associated with the effects of miR-451 on B-CPAP human PTC cells in vitro. MATERIAL AND METHODS Binding of miRNAs to the 3' untranslated region (3'UTR) of messenger RNA (mRNA) was determined with a luciferase reporter assay. miRNAs and plasmids were transfected into human PTC B-CPAP cells with Lipofectamine 2000 Transfection Reagent. Cell viability was tested with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay. The levels of miRNAs and mRNA were determined with quantitative polymerase chain reaction and protein levels were analyzed with immunoblotting. RESULTS miR-451 bound to wild-type but not mutant 3'-UTR of activating transcription factor 2 (ATF2). MiR-451 mimics inhibited the growth of B-CPAP cells and reduced mRNA and protein levels in ATF2, whereas miR-451 inhibitors promoted the growth of B-CPAP cells and increased mRNA and protein levels in ATF2. CONCLUSIONS miR-451 directly bound to the 3'UTR of ATF2, decreased mRNA and protein levels in ATF2, and inhibited growth of B-CPAP cells. Our findings suggest that miR-451 may be a potential therapeutic target for PTC.
MicroRNAs (miRNAs) 是肿瘤发生和癌症治疗耐药性中的重要新型生物标志物。miR-451 在人甲状腺乳头状癌 (PTC) 组织中表达,并与肿瘤进展相关。本研究探讨了 miR-451 对体外 B-CPAP 人 PTC 细胞影响的相关分子机制。
通过荧光素酶报告基因检测确定 miRNA 与信使 RNA (mRNA) 3'非翻译区 (3'UTR) 的结合。用 Lipofectamine 2000 Transfection Reagent 将 miRNA 和质粒转染入人 PTC B-CPAP 细胞。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐 (MTT) 法检测细胞活力。用定量聚合酶链反应 (qPCR) 测定 miRNA 和 mRNA 的水平,用免疫印迹法分析蛋白水平。
miR-451 与野生型但非突变型激活转录因子 2 (ATF2) 的 3'-UTR 结合。miR-451 模拟物抑制 B-CPAP 细胞的生长,并降低 ATF2 的 mRNA 和蛋白水平,而 miR-451 抑制剂促进 B-CPAP 细胞的生长,并增加 ATF2 的 mRNA 和蛋白水平。
miR-451 直接与 ATF2 的 3'UTR 结合,降低 ATF2 的 mRNA 和蛋白水平,并抑制 B-CPAP 细胞的生长。我们的研究结果表明,miR-451 可能是 PTC 的一个潜在治疗靶点。