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揭示miRNA30b在抑制ADAM12以对抗三阴性乳腺癌中的作用。

Unveiling miRNA30b's Role in Suppressing ADAM12 to Combat Triple-Negative Breast Cancer.

作者信息

Yin Qing-Hua, Hu Jian-Bing, Zhou Qiang, Weng Jie, Shen Er-Dong, Wen Fang, Liu Song-Lian, Yin Lei-Lan, Tong Ya-Jun, Long Ling, Tang Ke-Wei, Bai Si-Te, Ou Lu-di

机构信息

Department of Oncology, Yueyang Central Hospital, Yueyang 414000, Hunan, China.

出版信息

Breast J. 2024 Oct 30;2024:5202941. doi: 10.1155/2024/5202941. eCollection 2024.

Abstract

Triple-negative breast cancer, a subtype of breast cancer, is characterized by a poor prognosis. Recent studies have shown that miRNA30b acts as an oncogene and is vital for the proliferation of malignancies across various systems. This study aimed to elucidate the impact of miRNA30b on the proliferation, migration, and invasion capabilities of breast cancer cells . Triple-negative breast cancer cell lines MDA-MB-231 were transiently transfected with miRNA30b inhibitor, mimic, or the negative control by Lipofectamine 2000. Successful transfection was confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Functional assays, including CCK8, clone formation, scratch, and transwell assays, were conducted to evaluate the proliferation, invasion, and migration ability of MDA-MB-231 cells in each group. The target protein of miRNA30b was determined using an online prediction data website, and the dual-luciferase assay confirmed whether there was a binding site between miRNA30b and ADAM12. The effect was further verified by Western blot analysis. MDA-MB-231 cells were transfected with miRNA30b inhibitor, mimic, and negative control. miRNA30b expression was downregulated in the cells. Relative to the negative control group, miRNA30b expression significantly increased in the mimic group and decreased in the miRNA30b inhibitor group, with the differences being statistically significant. The miRNA30b mimic group exhibited a significant increase in miRNA30b expression and a corresponding promotion of cell proliferation, colony formation, and migration. Conversely, the miRNA30b inhibitor group displayed significantly reduced miRNA30b expression and suppressed cell proliferation, colony formation, and migration abilities compared to the negative control cells. Bioinformatics software predicted ADAM12 as a potential target of miRNA30b. Dual-luciferase assays confirmed the presence of a binding site between miRNA30b and ADAM12. Western blot analysis revealed that overexpression of miRNA30b downregulated ADAM12 expression in MDA-MB-231 cells. miRNA30b could promote proliferation, migration, and invasion of TNBC cell lines MDA-MB-231. The effect of miRNA30b on triple-negative breast cancer would be achieved partly at least through inhibiting the expression of ADAM12.

摘要

三阴性乳腺癌是乳腺癌的一种亚型,其特征是预后较差。最近的研究表明,miRNA30b作为一种癌基因,对各种系统中恶性肿瘤的增殖至关重要。本研究旨在阐明miRNA30b对乳腺癌细胞增殖、迁移和侵袭能力的影响。通过Lipofectamine 2000将miRNA30b抑制剂、模拟物或阴性对照瞬时转染到三阴性乳腺癌细胞系MDA-MB-231中。通过定量实时聚合酶链反应(qRT-PCR)确认转染成功。进行了包括CCK8、克隆形成、划痕和Transwell实验在内的功能实验,以评估每组中MDA-MB-231细胞的增殖、侵袭和迁移能力。使用在线预测数据网站确定miRNA30b的靶蛋白,双荧光素酶实验证实miRNA30b与ADAM12之间是否存在结合位点。通过蛋白质免疫印迹分析进一步验证其作用效果。用miRNA30b抑制剂、模拟物和阴性对照转染MDA-MB-231细胞。细胞中miRNA30b表达下调。相对于阴性对照组,模拟物组中miRNA30b表达显著增加,而miRNA30b抑制剂组中miRNA30b表达降低,差异具有统计学意义。miRNA30b模拟物组中miRNA30b表达显著增加,相应地促进了细胞增殖、集落形成和迁移。相反,与阴性对照细胞相比,miRNA30b抑制剂组中miRNA30b表达显著降低,细胞增殖、集落形成和迁移能力受到抑制。生物信息学软件预测ADAM12是miRNA30b的潜在靶标。双荧光素酶实验证实miRNA30b与ADAM12之间存在结合位点。蛋白质免疫印迹分析显示,miRNA30b的过表达下调了MDA-MB-231细胞中ADAM12的表达。miRNA30b可促进三阴性乳腺癌细胞系MDA-MB-231的增殖、迁移和侵袭。miRNA30b对三阴性乳腺癌的作用至少部分是通过抑制ADAM12的表达来实现的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d4b/11540880/906dbf051062/TBJ2024-5202941.001.jpg

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