Department of Colorectal Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, No. 44 Xiaoheyan Road, Dadong District, Shenyang city, Liaoning Province, China.
Biochem Genet. 2023 Jun;61(3):861-878. doi: 10.1007/s10528-022-10287-4. Epub 2022 Oct 8.
A large number of studies have shown that circular RNAs (circRNAs) are of great significance in the occurrence and development of colorectal cancer (CRC). The purpose of this study was to explore the mechanism of circ_0001535 in CRC. The expressions of circ_0001535, miR-433-3p and recombination signal-binding protein Jκ (RBPJ) mRNA and protein in CRC tissues and cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot. The effect of circ_0001535 on cell proliferation was detected using the Cell Counting Kit-8 (CCK-8) assay, colony formation assay and 5-ethynyl-2'-deoxyuridine (EdU) assay. The effects of circ_0001535 on migration, invasion, angiogenesis and apoptosis were investigated by wound healing assay, transwell assay, tube formation assay and flow cytometry, respectively. The interactions between miR-433-3p and circ_0001535 or RBPJ were studied using dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Xenograft tumor assay was performed to verify the role of circ_0001535 in tumor growth in vivo. The results showed that circ_0001535 and RBPJ mRNA expression levels were up-regulated and miR-433-3p was down-regulated in CRC tissues and cells. Circ_0001535 knockdown inhibited cell proliferation, migration, invasion, angiogenesis as well as promoted apoptosis in CRC cells. After analysis, it was found that circ_0001535 acted as a competing endogenous RNA (ceRNA) to inhibit miR-433-3p and then up-regulate RBPJ in CRC cells. In addition, in vivo experiment had shown that circ_0001535 knockdown inhibited tumor growth by up-regulating miR-433-3p and inhibiting RBPJ expression. The circ_0001535/miR-433-3p/ RBPJ axis plays a catalytic role in the progression of CRC, which may provide new insights into the molecular mechanism of CRC.
大量研究表明,环状 RNA(circRNA)在结直肠癌(CRC)的发生和发展中具有重要意义。本研究旨在探讨 circ_0001535 在 CRC 中的作用机制。采用实时定量聚合酶链反应(qRT-PCR)或 Western blot 检测 CRC 组织和细胞中 circ_0001535、miR-433-3p 和重组信号结合蛋白 Jκ(RBPJ)mRNA 和蛋白的表达。采用细胞计数试剂盒-8(CCK-8)检测 circ_0001535 对细胞增殖的影响,采用集落形成实验和 5-乙炔基-2'-脱氧尿苷(EdU)实验检测细胞增殖的影响。通过划痕愈合实验、Transwell 实验、管形成实验和流式细胞术分别研究 circ_0001535 对迁移、侵袭、血管生成和凋亡的影响。采用双荧光素酶报告基因实验和 RNA 免疫沉淀(RIP)实验研究 miR-433-3p 与 circ_0001535 或 RBPJ 的相互作用。通过异种移植肿瘤实验验证 circ_0001535 在体内肿瘤生长中的作用。结果表明,CRC 组织和细胞中 circ_0001535mRNA 表达上调,miR-433-3p 表达下调,RBPJmRNA 表达下调。circ_0001535 敲低抑制 CRC 细胞的增殖、迁移、侵袭和血管生成,并促进细胞凋亡。分析发现,circ_0001535 在 CRC 细胞中作为竞争性内源性 RNA(ceRNA)抑制 miR-433-3p,然后上调 RBPJ。此外,体内实验表明,circ_0001535 敲低通过上调 miR-433-3p 和抑制 RBPJ 表达抑制肿瘤生长。circ_0001535/miR-433-3p/RBPJ 轴在 CRC 进展中起催化作用,可能为 CRC 的分子机制提供新的见解。