Department of Brest, Affiliated Hospital of Hebei Engineering University, Handan, Hebei, China.
Department of Hepatobiliary Surgery, Handan Central Hospital, Handan, Hebei, China.
Bioengineered. 2022 Feb;13(2):2611-2622. doi: 10.1080/21655979.2021.2019877.
This work is designed to probe the functions and mechanisms of circ_0000518 in breast cancer (BC). qRT-PCR was performed to evaluate the circ_0000518, miR-1225-3p and Sry‑Related HMG box 4 (SOX4) mRNA expression in BC tissues and cells. After circ_0000518 was overexpressed in MDA-MB-468 cells, and circ_0000518 was knocked down in BT549 cells, CCK-8 test, and EdU assay were performed to measure the viability and growth of MDA-MB-468 and BT549 cells. Wound healing experiment was executed to determine the migration of BC cells. The invasion of cells was studied by the Transwell assay. Bioinformatics analysis, dual-luciferase reporter gene assay, qRT-PCR and Western blot were applied to predict and verify the binding sites between circ_0000518 and miR-1225-3p, miR-1225-3p and SOX4 mRNA. Pearson's correlation analysis was utilized to evaluate the correlations among circ_0000518 expression, miR-1225-3p expression, and SOX4 mRNA expression in BC specimens. It was revealed that, circ_0000518 and SOX4 mRNA expression levels were up-modulated in BC tissues, while miR-1225-3p expression was down-modulated in BC tissues than that in adjacent tissues. Circ_0000518 overexpression or inhibition of miR-1225-3p remarkably enhanced the growth, migration as well as invasion of BC cells , whereas circ_0000518 knockdown or miR-1225-3p overexpression worked oppositely. Circ_0000518 was identified as a molecular sponge of miR-1225-3p, and it can up-regulate SOX4 mRNA expression via repressing miR-1225-3p. In conclusion, circ_0000518 is oncogenic in BC and functions through miR-1225-3p/SOX4 axis.
这项工作旨在探究 circ_0000518 在乳腺癌（BC）中的功能和机制。通过 qRT-PCR 评估了 BC 组织和细胞中的 circ_0000518、miR-1225-3p 和 Sry-related HMG box 4（SOX4）mRNA 的表达。在 MDA-MB-468 细胞中转染 circ_0000518 过表达，BT549 细胞中转染 circ_0000518 敲低后，通过 CCK-8 试验和 EdU 测定法来测量 MDA-MB-468 和 BT549 细胞的活力和生长情况。通过划痕愈合实验来确定 BC 细胞的迁移情况。通过 Transwell 测定法来研究细胞的侵袭。通过生物信息学分析、双荧光素酶报告基因检测、qRT-PCR 和 Western blot 来预测和验证 circ_0000518 与 miR-1225-3p、miR-1225-3p 与 SOX4 mRNA 之间的结合位点。Pearson 相关分析用于评估 BC 标本中 circ_0000518 表达、miR-1225-3p 表达和 SOX4 mRNA 表达之间的相关性。结果表明，circ_0000518 和 SOX4 mRNA 的表达水平在 BC 组织中上调，而 miR-1225-3p 的表达水平在 BC 组织中下调。circ_0000518 过表达或抑制 miR-1225-3p 显著增强了 BC 细胞的生长、迁移和侵袭，而 circ_0000518 敲低或 miR-1225-3p 过表达则相反。circ_0000518 被鉴定为 miR-1225-3p 的分子海绵，通过抑制 miR-1225-3p 可上调 SOX4 mRNA 的表达。总之，circ_0000518 在 BC 中具有致癌作用，通过 miR-1225-3p/SOX4 轴发挥作用。
