Zhang Chao, Yang Ting
Department of Breast Surgery, Beijing Chaoyang Hospital Affiliated to Capital Medical University, Beijing 100020, China.
Department of Operating Room, New Century Women's and Children's Hospital, Beijing 100102, China.
Protein Pept Lett. 2023;30(1):72-84. doi: 10.2174/0929866530666221026164454.
There has been a large increase in the incidence of breast cancer (BC) among women. LINC00473 is a cancer-related lncRNA, participating in the progression of many cancers, but its role in the progression of BC awaits more elaboration.
Quantitative real-time polymerase chain reaction (qRT-PCR) was used to quantify LINC00473, miR-424-5p, and cyclin E1 (CCNE1) mRNA expression levels in BC tissues and cells. Cell counting kit-8 (CCK-8) assay was employed to detect the cell viability; the cell migration and invasion abilities were evaluated by the Transwell assay. Western blot and immunohistochemistry (IHC) were adopted to study CCNE1 protein expression; dual-luciferase reporter assay was performed to clarify the targeting relationships among LINC00473, miR-424-5p, and CCNE1.
LINC00473 expression was elevated in BC tissues and cell lines, which was associated with lymph node metastasis and higher clinical stage of the patients with BC. LINC00473 proved to be a molecular sponge for miR-424-5p; LINC00473 knockdown impeded the growth, migration, invasion, and epithelial-mesenchymal transition of BC cells, while these effects were abolished by miR-424-5p inhibitors; miR-424-5p targeted CCNE1 to restrain its expression. LINC00473 positively regulated CCNE1 expression, and CCNE1 restoration counteracted the effects induced by LINC00473 knockdown in BC cells.
LINC00473 facilitates the progression of BC through miR-424-5p/CCNE1 axis.
女性乳腺癌(BC)的发病率大幅上升。LINC00473是一种与癌症相关的长链非编码RNA(lncRNA),参与多种癌症的进展,但其在BC进展中的作用尚待进一步阐明。
采用定量实时聚合酶链反应(qRT-PCR)来定量BC组织和细胞中LINC00473、miR-424-5p和细胞周期蛋白E1(CCNE1)的mRNA表达水平。使用细胞计数试剂盒-8(CCK-8)检测细胞活力;通过Transwell实验评估细胞迁移和侵袭能力。采用蛋白质免疫印迹法(Western blot)和免疫组织化学(IHC)研究CCNE1蛋白表达;进行双荧光素酶报告基因实验以阐明LINC00473、miR-424-5p和CCNE1之间的靶向关系。
LINC00473在BC组织和细胞系中表达升高,这与BC患者的淋巴结转移及更高的临床分期相关。LINC00473被证明是miR-424-5p的分子海绵;敲低LINC00473可抑制BC细胞的生长、迁移、侵袭及上皮-间质转化,而miR-424-5p抑制剂可消除这些作用;miR-424-5p靶向CCNE1以抑制其表达。LINC00473正向调节CCNE1的表达,恢复CCNE1可抵消BC细胞中敲低LINC00473所诱导的作用。
LINC00473通过miR-424-5p/CCNE1轴促进BC的进展。