Department of Thyroid and Breast Surgery, The Affiliated Hospital of Jianghan University, Wuhan Hubei, China.
Department of Oncology, The Affiliated Hospital of Jianghan University, Wuhan Hubei, China.
Bioengineered. 2021 Dec;12(2):12394-12406. doi: 10.1080/21655979.2021.2005217.
Breast cancer, with high morbidity worldwide, is a threat to the life of women. MiR-543 was identified as playing an active part in the development of breast cancer involving multiple molecules. The goal of this study was to explore the molecular mechanisms of the involvement of miR-543 in the development of breast cancer. Quantitative real-time PCR (qRT-PCR) or Western blotting was used to detect mRNA or protein expression. Cell counting kit-8 (CCK-8), and the 5-bromo-2'-deoxyuridine (BrdU), wound healing, and Transwell assays were the main experimental procedures. Furthermore, subcutaneous tumor formation experiments were conducted to detect the function of miR-543 in breast cancer development in vivo. The match of miR-543 and ubiquitin-conjugating enzyme E2T (UBE2T) was detected through a dual-luciferase reporter experiment and RNA pull-down assay. Based on these results, miR-543 exhibited reduced expression in breast cancer tissues and cell lines, whereas UBE2T exhibited high levels. Furthermore, miR-543 directly targeted UBE2T, and a negative correlation between miR-543 and UBE2T was also observed in breast cancer tissues. Moreover, miR-543 overexpression led to inhibition of viability, proliferation, migration, and invasion of breast cancer. Furthermore, miR-543 overexpression undermined the UBE2T promotional effect by inhibiting ERK/MAPK pathway activity in breast cancer cells. Our study revealed that miR-543 impaired breast cancer progression by targeting UBE2T and downregulating UBE2T expression through the ERK/MAPK pathway, which suggested that miR-543 and UBE2T might serve as promising therapeutic gene targets for breast cancer in clinical application.
乳腺癌在全球范围内发病率高,严重威胁女性生命。miR-543 被认为在涉及多种分子的乳腺癌发生发展中起积极作用。本研究旨在探讨 miR-543 参与乳腺癌发生发展的分子机制。采用实时定量 PCR(qRT-PCR)或 Western blot 检测 mRNA 或蛋白表达。细胞计数试剂盒-8(CCK-8)、5-溴-2'-脱氧尿苷(BrdU)、划痕愈合和 Transwell 检测是主要的实验程序。此外,还进行了皮下肿瘤形成实验,以检测 miR-543 在体内乳腺癌发展中的功能。通过双荧光素酶报告实验和 RNA 下拉实验检测 miR-543 与泛素结合酶 E2T(UBE2T)的匹配。结果表明,miR-543 在乳腺癌组织和细胞系中表达降低,而 UBE2T 表达升高。此外,miR-543 直接靶向 UBE2T,并且在乳腺癌组织中也观察到 miR-543 和 UBE2T 之间呈负相关。此外,miR-543 过表达导致乳腺癌细胞活力、增殖、迁移和侵袭受到抑制。此外,miR-543 过表达通过抑制 ERK/MAPK 通路活性破坏乳腺癌细胞中 UBE2T 的促进作用。本研究表明,miR-543 通过靶向 UBE2T 并通过 ERK/MAPK 通路下调 UBE2T 表达来抑制乳腺癌进展,这表明 miR-543 和 UBE2T 可能成为临床应用中乳腺癌有前途的治疗基因靶点。