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miR-335-5p 通过靶向 SDC1 调控乳腺癌的进展。

miR-335-5p Targets SDC1 to Regulate the Progression of Breast Cancer.

机构信息

Department of Oncology, Shengjing Hospital of China Medical University, Shenyang, 110004, China.

出版信息

Crit Rev Eukaryot Gene Expr. 2022;32(6):21-31. doi: 10.1615/CritRevEukaryotGeneExpr.2022041813.

Abstract

The objective of the study was to explore the role of SDC1 in breast cancer cells. Our study also investigated the regulatory relationship between SDC1 and the microRNA (miRNA) miR-335-5p as well as the impact of these two genes on the progression of breast cancer. Bioinformatic approaches were employed to analyze the differentially expressed messenger RNAs (mRNAs) and miRNAs (DE-mRNAs and DE-miRNAs) in breast cancer tissue. Then mRNA SC1 was obtained. Differentially downregulated mRNAs were intersected with target miRNAs predicted by databases, and miR-335-5p was determined as the study object. Quantitative reverse transcription polymerase chain reaction was applied to assess the expressions of SDC1 and miR-335-5p in each cell line. Next, Western blot assay was conducted to detect the protein level of SDC1 and dual-luciferase assay was performed to verify the binding relationship between miR-335-5p and SDC1. Finally, we conducted methyl thiazolyl tetrazolium (MTT), colony formation, and Transwell assays and flow cytometry to further investigate the impacts of SDC1 and miR-335-5p on the progression of breast cancer. SDC1 was significantly highly expressed while miR-335-5p was remarkably lowly expressed in human breast cancer. Silencing SDC1 in breast cancer blocked the proliferation, migration and invasion of the cells. In breast cancer, SDC1 was a target gene of miR-335-5p and silencing miR-335-5p notably increased SDC1 expression. Compared with the silence of miR-335-5p, simultaneous silences of miR-335-5p and SDC1 significantly reduced the proliferative, migratory and invasive abilities of breast cancer cells. The result revealed the interaction between miR-335-5p and SDC1 in the progression of breast cancer, which may contribute to the treatments for this cancer.

摘要

本研究旨在探讨 SDC1 在乳腺癌细胞中的作用。我们的研究还探讨了 SDC1 与 microRNA (miRNA) miR-335-5p 之间的调控关系,以及这两个基因对乳腺癌进展的影响。采用生物信息学方法分析乳腺癌组织中差异表达的信使 RNA (mRNA) 和 miRNA (DE-mRNA 和 DE-miRNA)。然后获得 mRNA SC1。将数据库预测的靶 miRNA 与差异下调的 mRNA 进行交集,确定 miR-335-5p 为研究对象。采用实时定量逆转录聚合酶链反应 (qRT-PCR) 评估各细胞系中 SDC1 和 miR-335-5p 的表达。接下来,采用 Western blot 检测 SDC1 的蛋白水平,采用双荧光素酶报告基因检测验证 miR-335-5p 与 SDC1 的结合关系。最后,我们通过噻唑蓝(MTT)比色法、集落形成实验、Transwell 实验和流式细胞术进一步研究 SDC1 和 miR-335-5p 对乳腺癌进展的影响。在人乳腺癌中,SDC1 显著高表达,而 miR-335-5p 显著低表达。沉默乳腺癌中的 SDC1 可阻断细胞的增殖、迁移和侵袭。在乳腺癌中,SDC1 是 miR-335-5p 的靶基因,沉默 miR-335-5p 显著增加 SDC1 的表达。与沉默 miR-335-5p 相比,同时沉默 miR-335-5p 和 SDC1 显著降低了乳腺癌细胞的增殖、迁移和侵袭能力。结果揭示了 miR-335-5p 和 SDC1 在乳腺癌进展中的相互作用,可能有助于该癌症的治疗。

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