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微小 RNA-451 通过下调激活转录因子 2 的表达来减少 B-CPAP 人甲状腺乳头状癌细胞的增殖。

Micro-RNA-451 Reduces Proliferation of B-CPAP Human Papillary Thyroid Cancer Cells by Downregulating Expression of Activating Transcription Factor 2.

机构信息

Department of General Surgery, Chongming Branch of Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, China (mainland).

出版信息

Med Sci Monit. 2021 Mar 16;27:e929774. doi: 10.12659/MSM.929774.

Abstract

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.

摘要

背景

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。

材料与方法

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。

结果

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。

结论

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2d7/7980498/75e109f355f5/medscimonit-27-e929774-g001.jpg

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