Department of Genernal Surgery, The Affiliated Wuxi NO.2 People's Hospital of Nanjing Medical University, Wuxi, 214002, China.
Department of Orthopaedics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China.
Biomed Pharmacother. 2020 May;125:109876. doi: 10.1016/j.biopha.2020.109876. Epub 2020 Feb 7.
There is increasing evidence of the vital role played by circular RNAs (circRNAs) in the progression of gastric cancer (GC). A circRNA, hsa_circ_0001772, was generated from the RBM33 gene and named circRBM33. The aim of this study was to investigate the role of circRBM33 in GC. Quantitative real-time PCR (qRT-PCR) was used to quantify the expression of circRBM33 in 79 pairs of GC tissues and paracancerous tissues and 4 GC cell lines (MGC-803, BGC-823, SGC-7901, and AGS). Bioinformatics databases were used to predict downstream targets of circRNA and micro RNA (miRNA). Dual luciferase reporter assay was used to verify whether miR-149 was a direct binding target for circRBM33. Cell Counting Kit-8 (CCK-8) assay, 5-Ethynyl-2´-deoxyuridine (EDU) assay, transwell assay, and flow-cytometric analyses were performed to determine the role of circRBM33 in the biological functioning of GC cells. Western blot technique was used to quantify the levels of interleukin-6 (IL-6). CircRBM33 was distinctly upregulated in GC specimens and cell lines and a close correlation between circRBM33 expression and clinical characteristics of GC was observed. After silencing circRBM33, the apoptosis of GC cells increased, while proliferation, migration, and invasion decreased. Rescue experiments indicated that circRBM33 manipulates biological function in GC cells through the circRBM33/miR-149/IL-6 axis. CircRBM33 can be used as a tumor biomarker and a possible therapeutic target in the future.
越来越多的证据表明,环状 RNA(circRNAs)在胃癌(GC)的进展中起着至关重要的作用。环状 RNA hsa_circ_0001772 来源于 RBM33 基因,并被命名为 circRBM33。本研究旨在探讨 circRBM33 在 GC 中的作用。采用定量实时 PCR(qRT-PCR)检测 79 对 GC 组织和癌旁组织及 4 种 GC 细胞系(MGC-803、BGC-823、SGC-7901 和 AGS)中 circRBM33 的表达。生物信息学数据库用于预测环状 RNA 和 micro RNA(miRNA)的下游靶标。双荧光素酶报告基因实验用于验证 miR-149 是否为 circRBM33 的直接结合靶标。细胞计数试剂盒-8(CCK-8)检测、5-乙炔基-2'-脱氧尿苷(EDU)检测、Transwell 检测和流式细胞术分析用于确定 circRBM33 在 GC 细胞生物学功能中的作用。Western blot 技术用于定量白细胞介素 6(IL-6)的水平。circRBM33 在 GC 标本和细胞系中明显上调,circRBM33 的表达与 GC 的临床特征密切相关。沉默 circRBM33 后,GC 细胞的凋亡增加,而增殖、迁移和侵袭减少。挽救实验表明,circRBM33 通过 circRBM33/miR-149/IL-6 轴调控 GC 细胞的生物学功能。circRBM33 可作为未来肿瘤的生物标志物和潜在的治疗靶点。