Xiao Weisheng, Li Peiyuan
Department of Gastroenterology, The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang 421001, China.
Department of Gastroenterology, The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang 421001, China.
Pathol Res Pract. 2023 Aug;248:154695. doi: 10.1016/j.prp.2023.154695. Epub 2023 Jul 17.
Circular RNAs (circRNAs) exert crucial roles in tumor progression of multiple cancers, including colorectal cancer (CRC). However, the functions of most circRNAs are not been fully elucidated. In this study, the role and mechanism of circ_0087862 in CRC were investigated.
The expression of circ_0087862, microRNA-296-3p (miR-296-3p) and phosphoglycerate kinase 1 (PGK1) was detected by quantitative real-time PCR (qRT-PCR). Cell Counting Kit-8 (CCK-8) assay and 5-ethynyl-2'-deoxyuridine (EdU) assay were used to assess cell proliferation. Flow cytometry was employed to analyze cell apoptosis. Transwell assay was employed to evaluate cell invasion. Western blot assay was employed to detect the level of related protein markers and PGK1. The glucose consumption, lactate production were tested by corresponding kits. The relationship between miR-296-3p and circ_0087862 or PGK1 was verified by dual-luciferase reporter assay or RNA immunoprecipitation (RIP) assay. The in vivo function of circ_0087862 was examined by xenograft mice model.
The expression levels of circ_0087862 and PGK1 were up-regulated in CRC tissues and cells, while miR-296-3p was down-regulated. Circ_0087862 silencing suppressed cell proliferation, invasion and glycolysis and promoted cell apoptosis in CRC cells. Circ_0087862 targeted miR-296-3p in CRC cells. MiR-296-3p inhibition reversed circ_0087862 silencing-mediated inhibition effect on cell proliferation, invasion and glycolysis, as well as the promotion effect on cell apoptosis. PGK1 was a target of miR-296-3p, and the overexpression of PGK1 attenuated miR-296-3p-mediated tumor suppression effect on CRC progression. Moreover, knockdown of circ_0087862 inhibited tumorigenesis in vivo.
Circ_0087862 promoted CRC progression via miR-296-3p/PGK1 axis and might act as a potential target for CRC therapy.
环状RNA（circRNAs）在包括结直肠癌（CRC）在内的多种癌症的肿瘤进展中发挥关键作用。然而，大多数circRNAs的功能尚未完全阐明。在本研究中，我们探究了circ_0087862在结直肠癌中的作用及机制。
采用定量实时荧光定量PCR（qRT-PCR）检测circ_0087862、微小RNA-296-3p（miR-296-3p）和磷酸甘油酸激酶1（PGK1）的表达。使用细胞计数试剂盒-8（CCK-8）法和5-乙炔基-2'-脱氧尿苷（EdU）法评估细胞增殖。采用流式细胞术分析细胞凋亡。采用Transwell法评估细胞侵袭。采用蛋白质免疫印迹法检测相关蛋白标志物和PGK1的水平。通过相应试剂盒检测葡萄糖消耗和乳酸生成。通过双荧光素酶报告基因检测或RNA免疫沉淀（RIP）检测验证miR-296-3p与circ_0087862或PGK1之间的关系。通过异种移植小鼠模型检测circ_0087862的体内功能。
circ_0087862和PGK1在结直肠癌组织和细胞中的表达水平上调，而miR-296-3p下调。circ_0087862沉默抑制了结直肠癌细胞的增殖、侵袭和糖酵解，并促进细胞凋亡。circ_0087862在结直肠癌细胞中靶向miR-296-3p。miR-296-3p抑制逆转了circ_0087862沉默介导的对细胞增殖、侵袭和糖酵解的抑制作用，以及对细胞凋亡的促进作用。PGK1是miR-296-3p的靶点，PGK1的过表达减弱了miR-296-3p介导的对结直肠癌进展的肿瘤抑制作用。此外，circ_0087862的敲低抑制了体内肿瘤发生。
circ_0087862通过miR-296-3p/PGK1轴促进结直肠癌进展，可能作为结直肠癌治疗的潜在靶点。
