Department of Urology, The Affiliated Hospital of Guizhou Medical University, Guiyang, China,
Department of Urology, The Affiliated Hospital of Guizhou Medical University, Guiyang, China.
Pathobiology. 2020;87(6):345-355. doi: 10.1159/000509957. Epub 2020 Nov 25.
Long noncoding RNAs (lncRNAs) are potential biomarkers that are very important for the development of cancer. Studies show that lncRNAs are significantly correlated with the carcinogenesis and progression of bladder cancer (BLCA). In this research, we aimed at probing into the role of lncRNA MAFG-AS1 in the tumorigenesis of BLCA.
RT-qPCR was employed to detect MAFG-AS1 expression in BLCA tissues and cells. MAFG-AS1 siRNA and overexpression plasmid were transfected into 5637 and T24 BLCA cell lines to inhibit or upregulate MAFG-AS1 expression, respectively, and then the regulatory functions of MAFG-AS1 on BLCA cell proliferation, migration, and invasion were assessed using cell counting kit-8 (CCK-8) assay, EdU method, and Transwell experiments, respectively. Dual-luciferase reporter assay and RNA immunoprecipitation were conducted to validate the targeting relationships between MAFG-AS1 and miR-143-3p, and miR-143-3p and COX-2. In addition, miR-143-3p was repressed in MAFG-AS1-silenced 5637 and T24 cell lines, and the function of MAFG-AS1/miR-143-3p axis in BLCA cells was further evaluated. The regulatory effects of MAFG-AS1 and miR-143-3p on the expression of COX-2 protein were detected by Western blot.
MAFG-AS1 was remarkably upregulated in BLCA patient tissues and cell lines, and its high expression was closely related to histological grade, tumor size, and lymph node metastasis. Silencing of MAFG-AS1 inhibited BLCA cell proliferation, metastasis, and invasion, while overexpression of MAFG-AS1 in BLCA cells had opposite biological effects. MAFG-AS1 was proved to target miR-143-3p to repress its expression. Moreover, it was confirmed that MAFG-AS1 and miR-143-3p could modulate COX-2 expression.
The MAFG-AS1/miR-143-3p/COX-2 axis contributes to BLCA progression.
长链非编码 RNA（lncRNA）是癌症发生发展的重要潜在标志物。研究表明，lncRNA 与膀胱癌（BLCA）的发生和发展显著相关。在这项研究中，我们旨在探讨 lncRNA MAFG-AS1 在 BLCA 肿瘤发生中的作用。
采用 RT-qPCR 检测 BLCA 组织和细胞中 MAFG-AS1 的表达。用 MAFG-AS1 siRNA 和过表达质粒分别转染 5637 和 T24 BLCA 细胞系，抑制或上调 MAFG-AS1 表达，然后分别采用细胞计数试剂盒（CCK-8）法、EdU 法和 Transwell 实验评估 MAFG-AS1 对 BLCA 细胞增殖、迁移和侵袭的调控作用。采用双荧光素酶报告基因实验和 RNA 免疫沉淀实验验证 MAFG-AS1 与 miR-143-3p 及 miR-143-3p 与 COX-2 的靶向关系。此外，在沉默 MAFG-AS1 的 5637 和 T24 细胞系中转录抑制 miR-143-3p，并进一步评估 MAFG-AS1/miR-143-3p 轴在 BLCA 细胞中的作用。采用 Western blot 检测 MAFG-AS1 和 miR-143-3p 对 COX-2 蛋白表达的调控作用。
MAFG-AS1 在 BLCA 患者组织和细胞系中显著上调，其高表达与组织学分级、肿瘤大小和淋巴结转移密切相关。沉默 MAFG-AS1 抑制 BLCA 细胞增殖、转移和侵袭，而在 BLCA 细胞中过表达 MAFG-AS1 则具有相反的生物学效应。MAFG-AS1 被证实可靶向 miR-143-3p 抑制其表达。此外，证实 MAFG-AS1 和 miR-143-3p 可以调节 COX-2 表达。
MAFG-AS1/miR-143-3p/COX-2 轴参与 BLCA 进展。
