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

IF 1.9 4区 医学 Q3 OBSTETRICS & GYNECOLOGY Breast Journal Pub Date : 2024-10-30 DOI:10.1155/2024/5202941
Qing-hua Yin, Jian-bing Hu, Qiang Zhou, Jie Weng, Er-dong Shen, Fang Wen, Song-lian Liu, Lei-lan Yin, Ya-jun Tong, Ling Long, Ke-wei Tang, Si-te Bai, Lu-di Ou
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Abstract

Background: 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 in vitro.

Methods: 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.

Results: 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.

Conclusions: 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.

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揭示 miRNA30b 在抑制 ADAM12 对抗三阴性乳腺癌中的作用
背景:三阴性乳腺癌是乳腺癌的一种亚型,预后较差。最近的研究表明,miRNA30b 是一种致癌基因,对各系统恶性肿瘤的增殖至关重要。本研究旨在阐明 miRNA30b 对体外乳腺癌细胞增殖、迁移和侵袭能力的影响。 研究方法用 Lipofectamine 2000 给三阴性乳腺癌细胞株 MDA-MB-231 转染 miRNA30b 抑制剂、模拟物或阴性对照。转染成功与否由实时定量聚合酶链反应(qRT-PCR)确认。功能试验包括 CCK8、克隆形成、划痕和透孔试验,以评估各组 MDA-MB-231 细胞的增殖、侵袭和迁移能力。利用在线预测数据网站确定了 miRNA30b 的靶蛋白,并通过双荧光素酶试验证实了 miRNA30b 与 ADAM12 之间是否存在结合位点。通过 Western 印迹分析进一步验证了其效果。 结果用 miRNA30b 抑制剂、模拟物和阴性对照组转染 MDA-MB-231 细胞。与阴性对照组相比,miRNA30b表达在miRNA30b抑制剂组显著增加,在miRNA30b抑制剂组显著减少,差异有统计学意义。miRNA30b mimic 组的 miRNA30b 表达明显增加,相应地促进了细胞增殖、集落形成和迁移。相反,与阴性对照组相比,miRNA30b 抑制剂组的 miRNA30b 表达明显减少,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 的潜在靶点。双荧光素酶测定证实了 miRNA30b 与 ADAM12 之间存在结合位点。Western 印迹分析显示,过表达 miRNA30b 会降低 MDA-MB-231 细胞中 ADAM12 的表达。 结论:miRNA30b 可促进 TNBC 细胞株 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 的表达实现的。
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来源期刊
Breast Journal
Breast Journal 医学-妇产科学
CiteScore
4.00
自引率
0.00%
发文量
47
审稿时长
4-8 weeks
期刊介绍: The Breast Journal is the first comprehensive, multidisciplinary source devoted exclusively to all facets of research, diagnosis, and treatment of breast disease. The Breast Journal encompasses the latest news and technologies from the many medical specialties concerned with breast disease care in order to address the disease within the context of an integrated breast health care. This editorial philosophy recognizes the special social, sexual, and psychological considerations that distinguish cancer, and breast cancer in particular, from other serious diseases. Topics specifically within the scope of The Breast Journal include: Risk Factors Prevention Early Detection Diagnosis and Therapy Psychological Issues Quality of Life Biology of Breast Cancer.
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