Department of oncology, Peking Universtity Shenzhen Hospital, 1120 Lianhua Road, Futian District, Shenzhen, Guangdong Province, 518000, China.
Biochem Genet. 2024 Jun;62(3):1795-1810. doi: 10.1007/s10528-023-10522-6. Epub 2023 Sep 20.
Circular RNAs (circRNAs) closely related to the progression of colorectal cancer (CRC). Nevertheless, the study of circ_0005785 in CRC has not been reported. In this test, we aimed to investigate the mechanisms of circ_0005785 in CRC development. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were employed to reveal the expression of genes and proteins. Cell Counting Kit-8 (CCK-8) assay, 5-ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry analysis, transwell assay and tube formation experiment were implemented to examine cell growth, apoptosis, invasion and angiogenesis. The relationships among circ_0005785, miR-7-5p and DNA methyltransferase 3 A (DNMT3A) were verified by dual-luciferase reporter assay. Xenograft mouse model was built to evaluate the impacts of circ_0005785 deficiency on CRC growth in vivo. We found that circ_0005785 was increased in CRC patients and cell lines. Circ_0005785 downregulation retarded cell proliferation, invasion, angiogenesis whereas expedited apoptosis in CRC cells. Mechanistically, circ_0005785 could sponge miR-7-5p and the suppressive treads of circ_0005785 in CRC development was attenuated by miR-7-5p down-regulation. DNMT3A was targeted by miR-7-5p and miR-7-5p overexpression constrained cell malignant behaviors, but the addition of DNMT3A counteracted the effects. Additionally, circ_0005785 inhibition hindered the tumor growth in vivo. In conclusion, circ_0005785 aggravated the CRC progression by increasing the level of DNMT3A via adsorbing miR-7-5p.
环状 RNA(circRNAs)与结直肠癌(CRC)的进展密切相关。然而,circ_0005785 在 CRC 中的研究尚未报道。在本研究中,我们旨在探讨 circ_0005785 在 CRC 发生发展中的机制。采用实时定量聚合酶链反应(qRT-PCR)和 Western blot 检测基因和蛋白的表达。细胞计数试剂盒-8(CCK-8)检测、5-乙炔基-2'-脱氧尿苷(EdU)检测、流式细胞术分析、Transwell 检测和管形成实验用于检测细胞生长、凋亡、侵袭和血管生成。通过双荧光素酶报告基因实验验证 circ_0005785、miR-7-5p 和 DNA 甲基转移酶 3A(DNMT3A)之间的关系。建立异种移植小鼠模型评估 circ_0005785 缺失对 CRC 体内生长的影响。结果发现,CRC 患者和细胞系中 circ_0005785 表达增加。circ_0005785 下调可抑制 CRC 细胞的增殖、侵袭和血管生成,促进其凋亡。机制上,circ_0005785 可以吸附 miR-7-5p,circ_0005785 在 CRC 中的抑制作用可被 miR-7-5p 下调减弱。DNMT3A 是 miR-7-5p 的靶基因,miR-7-5p 过表达抑制细胞恶性行为,但加入 DNMT3A 可拮抗这种作用。此外,circ_0005785 抑制可抑制体内肿瘤生长。总之,circ_0005785 通过吸附 miR-7-5p 增加 DNMT3A 的水平,加重了 CRC 的进展。