Zhang Bo, Cao Wenbin, Liu Yang, Zhao Yongkui, Liu Chunhui, Sun Bingfu
Department of General Surgery, North China University of Science and Technology Affiliated Hospital, No.73 South Jianshe Road, Tangshan, 063000, Hebei, China.
Mol Cell Biochem. 2023 Mar;478(3):503-516. doi: 10.1007/s11010-022-04525-x. Epub 2022 Aug 2.
The purpose of this paper was to explore the role of circ_0056618 and associated mechanisms in colorectal cancer (CRC). The expression of circ_0056618, proline rich and Gla domain 4 (PRRG4) mRNA and miR-411-5p was measured by quantitative real-time PCR (qPCR).The protein levels of PRRG4 and epithelial-mesenchymal transition (EMT)-related markers were detected by western blot. Cell proliferation was assessed by cell counting kit-8, EdU, and colony formation assays. Cell migration and invasion were assessed by transwell assay. Cell apoptosis was detected by flow cytometry assay. The putative relationship between miR-411-5p and circ_0056618 or PRRG4 was verified by dual-luciferase reporter assay. The effects of circ_0056618 on tumor growth in vivo were determined by animal study. Circ_0056618 and PRRG4 was upregulated, while miR-411-5p was downregulated in CRC tumor tissues and cells. Circ_0056618 knockdown or PRRG4 knockdown inhibited CRC cell proliferation, migration/invasion, EMT, and survival. Circ_0056618 positively modulated PRRG4 expression by targeting miR-411-5p. MiR-411-5p absence or PRRG4 overexpression could rescue circ_0056618 knockdown-induced inhibition on proliferation, migration/invasion, and EMT in CRC cells. Animal assay showed circ_0056618 knockdown impeded tumor growth in vivo. Circ_0056618 promoted CRC growth and development by upregulating PRRG4 expression via competitively targeting miR-411-5p.
本文旨在探讨circ_0056618在结直肠癌（CRC）中的作用及相关机制。采用定量实时PCR（qPCR）检测circ_0056618、富含脯氨酸和Gla结构域4（PRRG4）mRNA及miR-411-5p的表达。通过蛋白质印迹法检测PRRG4蛋白水平及上皮-间质转化（EMT）相关标志物。采用细胞计数试剂盒-8、EdU和集落形成试验评估细胞增殖。通过Transwell试验评估细胞迁移和侵袭。采用流式细胞术检测细胞凋亡。通过双荧光素酶报告试验验证miR-411-5p与circ_0056618或PRRG4之间的假定关系。通过动物实验确定circ_0056618对体内肿瘤生长的影响。在CRC肿瘤组织和细胞中，circ_0056618和PRRG4上调，而miR-411-5p下调。敲低circ_0056618或PRRG4可抑制CRC细胞增殖、迁移/侵袭、EMT及存活。circ_0056618通过靶向miR-411-5p正向调节PRRG4表达。miR-411-5p缺失或PRRG4过表达可挽救circ_0056618敲低诱导的CRC细胞增殖、迁移/侵袭及EMT抑制。动物实验表明，敲低circ_0056618可抑制体内肿瘤生长。circ_0056618通过竞争性靶向miR-411-5p上调PRRG4表达，从而促进CRC的生长和发展。
