Gu She Qun, Luo Ji Hui, Yao Wen Xiu
The First Department of Medical Oncology, The First People's Hospital of Chenzhou, Southern Medical University, Chenzhou 423000, China.
The Second General Surgery Department for Carcinoma, The First People's Hospital of Chenzhou, Chenzhou 423000, China.
Math Biosci Eng. 2019 Nov 27;17(2):1428-1441. doi: 10.3934/mbe.2020073.
This study aimed to explore the regulatory mechanism of miR-139-5p on the biological characteristics of breast cancer cells by targeting Collagen type XI alpha 1 chain (COL11A1). GEO2R was used to identify the differentially expressed genes (DEGs) of breast cancer in GEO database. miRDB, miRanda and TargetScan databases were used to predict the miRNAs that regulate COL11A1. qRT-PCR was used to detect the expressions of COL11A1 and miR-139-5p in breast cancer cells, and western blot was used to detect the protein level of COL11A1. RNA binding protein immunoprecipitation assay was employed to test the targeted relationship between miR-139-5p and COL11A1, which was further verified by dual-luciferase reporter gene assay. CCK-8 assay was performed to detect the cell proliferation, and flow cytometry was carried out to examine the cell apoptosis. Moreover, western blot was used for the detection of caspase-3, Bax and Bcl-2 protein levels. A total of five DEGs were screened from the GEO database, of which COL11A1 was the only highly expressed in breast cancer. According to the database analysis, we predicted that miR-139-5p was much likely to target the expression of COL11A1. MiR-139-5p was poorly expressed in breast cancer cells and targeted inhibited COL11A1. Silencing COL11A1 or overexpressing miR-139-5p both could inhibit the proliferation and promote the apoptosis, while overexpressing the two factors simultaneously could reverse such effect. Overexpression of miR-139-5p inhibits the proliferation and promotes the apoptosis of breast cancer cells by inhibiting the expression of COL11A1.
本研究旨在通过靶向XI型胶原蛋白α1链（COL11A1）来探索miR-139-5p对乳腺癌细胞生物学特性的调控机制。利用GEO2R在GEO数据库中鉴定乳腺癌的差异表达基因（DEG）。使用miRDB、miRanda和TargetScan数据库预测调控COL11A1的miRNA。采用qRT-PCR检测乳腺癌细胞中COL11A1和miR-139-5p的表达，并用蛋白质印迹法检测COL11A1的蛋白水平。采用RNA结合蛋白免疫沉淀试验检测miR-139-5p与COL11A1之间的靶向关系，并通过双荧光素酶报告基因试验进一步验证。进行CCK-8试验检测细胞增殖情况，采用流式细胞术检测细胞凋亡情况。此外，用蛋白质印迹法检测caspase-3、Bax和Bcl-2蛋白水平。从GEO数据库中共筛选出5个DEG，其中COL11A1是唯一在乳腺癌中高表达的基因。根据数据库分析，我们预测miR-139-5p很可能靶向COL11A1的表达。miR-139-5p在乳腺癌细胞中低表达，并靶向抑制COL11A1。沉默COL11A1或过表达miR-139-5p均可抑制增殖并促进凋亡，而同时过表达这两个因子可逆转这种效应。过表达miR-139-5p通过抑制COL11A1的表达来抑制乳腺癌细胞的增殖并促进其凋亡。
