Department of Oncology, Shengjing Hospital of China Medical University, Shenyang, 110004, China.
Crit Rev Eukaryot Gene Expr. 2022;32(6):21-31. doi: 10.1615/CritRevEukaryotGeneExpr.2022041813.
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.
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。
