Liu Guanglan, Liu Xinli, Yin Junfeng, Zheng Haijian, Zhu Xinguo
Department of General Surgery, The First Affiliated Hospital of Soochow University, NO. 188 Shizi Street, Suzhou, 215006, Jiangsu, China.
Department of Digestive Oncology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, 44 Xiaoheyan Road, Shenyang, 110042, Liaoning, China.
Noncoding RNA Res. 2024 Mar 13;9(3):921-929. doi: 10.1016/j.ncrna.2024.03.007. eCollection 2024 Sep.
Colorectal cancer (CRC) incidence ranks third among malignant cancers with a high propensity for distant metastasis. Despite continuous efforts to improve treatment, the prognosis especially in patients with advanced distant metastasis is low. The mechanism of development and progression of CRC is not fully understood. Non-coding RNAs (ncRNAs) have emerged as essential regulators in cancer progression. Here, we aim to dissect the role of one critical ncRNA, circANXA4, in CRC progression. CircANXA4 expression was analyzed by the GEO database. Differentially expressed circRNAs were identified by the Limma package R software. Expression of circANXA4 and miR-1256 was detected by qRT-PCR. The regulation of circANXA4 on cell proliferation and progression was confirmed with the cell viability assay using cell counting kit-8 (CCK-8) and transwell migration assay. RNA pull-down assay, RNA immunoprecipitation (RIP), and western blot were used to determine the interaction between circANXA4, miR-1256, and protamine1 (PRM1). CircANXA4 was upregulated in both CRC tissues and cell lines. Knockdown of circANXA4 effectively reduced cell proliferation, progression, and migration. Additionally, silencing circANXA4 remarkably increased miR-1256 expression, while reducing PRM1 expression, thereby demonstrating that circANXA4 downregulates miR-1256 expression through a complementary binding site. Rescue experiments revealed the interactions between circANXA4, miR-1256, and PRM1. Pearson correlation analysis revealed that circANXA4 expression positively correlated with PRM1 expression and miR-1256 expression inversely correlated with PRM1 expression. In sum, we demonstrated that circANXA4 promotes cancer cell proliferation and progression by sponging miR-1256 and upregulating PRM1 in CRC.
结直肠癌（CRC）的发病率在恶性肿瘤中位居第三，具有较高的远处转移倾向。尽管在改善治疗方面不断努力，但预后尤其是晚期远处转移患者的预后仍然很差。CRC发生和发展的机制尚未完全阐明。非编码RNA（ncRNAs）已成为癌症进展中的重要调节因子。在这里，我们旨在剖析一种关键的ncRNA，即circANXA4在CRC进展中的作用。通过GEO数据库分析circANXA4的表达。使用Limma包R软件鉴定差异表达的circRNAs。通过qRT-PCR检测circANXA4和miR-1256的表达。使用细胞计数试剂盒-8（CCK-8）进行细胞活力测定和Transwell迁移测定，证实了circANXA4对细胞增殖和进展的调节作用。采用RNA下拉试验、RNA免疫沉淀（RIP）和蛋白质印迹法确定circANXA4、miR-1256和鱼精蛋白1（PRM1）之间的相互作用。circANXA4在CRC组织和细胞系中均上调。敲低circANXA4可有效降低细胞增殖、进展和迁移。此外，沉默circANXA4可显著增加miR-1256的表达，同时降低PRM1的表达，从而表明circANXA4通过互补结合位点下调miR-1256的表达。挽救实验揭示了circANXA4、miR-1256和PRM1之间的相互作用。Pearson相关性分析显示，circANXA4表达与PRM1表达呈正相关，而miR-1256表达与PRM1表达呈负相关。总之，我们证明了circANXA4通过在CRC中吸附miR-1256并上调PRM1来促进癌细胞增殖和进展。
