College of Life Sciences, Sichuan University, Chengdu 610065, China.
College of Life Sciences, Sichuan University, Chengdu 610065, China.
Biochim Biophys Acta Mol Basis Dis. 2023 Dec;1869(8):166821. doi: 10.1016/j.bbadis.2023.166821. Epub 2023 Jul 27.
Development of colorectal cancer (CRC) accompanied with genomic instability. Genomic instability was promoted by microRNAs (miRNAs) inhibiting key genes in DNA damage repair and spindle assembly processes. Whether miR-653-3p affects genomic instability is unknown. The aim of this study is to explore the effect of miR-653-3p on genomic instability in CRC cells. Based on RT-qPCR analysis, miR-653-3p was highly expressed in CRC cells. Through single-cell electrophoresis assay and chromosome karyotype analysis, we determined ectopic expression of miR-653-3p induced increased DNA damage but inhibited apoptosis by promoting chromosomal instability. Mechanistically, luciferase assay identified the direct interaction of miR-653-3p with the 3' UTR of SIRT1, and western blot analysis indicated miR-653-3p inhibited SIRT1 and then promoted STAT3 phosphorylation and TWIST1 expression. The results of karyotype analysis showed that the upregulation of SIRT1 and the downregulation of TWIST1 caused by the downregulation of miR-653-3p suppressed chromosomal instability. Additionally, our evidence showed that miR-653-3p promoted CRC cell proliferation, migration, and 5-FU resistance, and miR-653-3p induced the development of CRC in the xenograft mice model. Altogether, our evidence suggests that miR-653-3p regulates SIRT1/TWIST1 signaling pathway and plays an important role in promoting genomic instability, proliferation, migration, and chemoresistance of CRC cells, which may serve as a promising therapeutic target for CRC therapy.
结直肠癌（CRC）的发生伴随着基因组不稳定。基因组不稳定性是由 microRNAs（miRNAs）抑制 DNA 损伤修复和纺锤体组装过程中的关键基因所促进的。miR-653-3p 是否影响基因组不稳定性尚不清楚。本研究旨在探讨 miR-653-3p 对 CRC 细胞基因组不稳定性的影响。基于 RT-qPCR 分析，miR-653-3p 在 CRC 细胞中高表达。通过单细胞电泳实验和染色体核型分析，我们确定 miR-653-3p 的异位表达诱导了 DNA 损伤的增加，但通过促进染色体不稳定抑制了细胞凋亡。机制上，荧光素酶报告基因实验鉴定了 miR-653-3p 与 SIRT1 的 3'UTR 的直接相互作用，Western blot 分析表明 miR-653-3p 抑制了 SIRT1，从而促进了 STAT3 磷酸化和 TWIST1 表达。核型分析的结果表明，miR-653-3p 下调引起的 SIRT1 上调和 TWIST1 下调抑制了染色体不稳定。此外，我们的证据表明，miR-653-3p 促进了 CRC 细胞的增殖、迁移和 5-FU 耐药性，并且 miR-653-3p 诱导了异种移植小鼠模型中 CRC 的发展。总之，我们的证据表明，miR-653-3p 通过调节 SIRT1/TWIST1 信号通路在促进 CRC 细胞的基因组不稳定、增殖、迁移和化疗耐药性方面发挥着重要作用，这可能成为 CRC 治疗的有前途的治疗靶点。
