Physical Examination Center, The First Affiliated Hospital of Jinzhou Medical University, No.2, Section 5, Renmin Street, Guta District, Jinzhou City, 121000, Liaoning Province, China.
Department of General Surgery, The First Affiliated Hospital of Jinzhou Medical University, No.2, Section 5, Renmin Street, Guta District, Jinzhou City, 121000, Liaoning Province, China.
J Bioenerg Biomembr. 2021 Oct;53(5):609-620. doi: 10.1007/s10863-021-09907-2. Epub 2021 Jun 23.
Thyroid carcinoma (TC) seriously threatens the health and safety of patients, and the treatment target of it still is poor. RT-qPCR and Western blot were carried out to detect the expression of genes and proteins, respectively. Cell proliferation was confirmed using colony formation assay. Transwell assay were performed to measure the cell migration and invasion. Besides, luciferase reporter assay was accomplished to ensure the target relationship between miR-942-5p and TWIST1 mRNA as well as hsa_circ_0001681. Here, we proved that hsa_circ_0001681 was increased in TC, and located majorly in the cytoplasm of TC cells. However, miR-942-5p was decreased in TC, and was negatively correlated with hsa_circ_0001681 expression. Knockdown of hsa_circ_0001681 significantly repressed the proliferation, migration, invasion and EMT of TC cells. We also found that the process of hsa_circ_0001681 silencing limited EMT, which was obstructed by TWIST1 increasing. Moreover, hsa_circ_0001681 acted as a miRNA sponge and completed with TWIST1 mRNA for binding to miR-942-5p, thus downregulation of hsa_circ_0001681 repressed EMT and subsequent malignant phenotype of TC cells through targeting miR-942-5p/TWIST1 signaling pathway. Finally, the studies in vivo showed that decreasing of hsa_circ_0001681 effectively inhibited the growth of tumor via repressing EMT by regulating miR-942-5p/TWIST1 signaling pathway. Overall, silencing of hsa_circ_0001681 significantly suppressed TC progression through inhibiting EMT via acting as a miR-942-5p sponge to facilitate the expression of TWIST1. Our data provided a reliable evidence for hsa_circ_0001681 is a potential treatment target in TC.
甲状腺癌(TC)严重威胁着患者的健康和安全,其治疗靶点仍不理想。分别采用 RT-qPCR 和 Western blot 检测基因和蛋白的表达。通过集落形成实验证实细胞增殖。通过 Transwell 实验测量细胞迁移和侵袭。此外,通过荧光素酶报告实验证实了 miR-942-5p 与 TWIST1 mRNA 以及 hsa_circ_0001681 的靶关系。在这里,我们证明了 hsa_circ_0001681 在 TC 中增加,并且主要位于 TC 细胞的细胞质中。然而,miR-942-5p 在 TC 中减少,并且与 hsa_circ_0001681 表达呈负相关。hsa_circ_0001681 的敲低显着抑制 TC 细胞的增殖、迁移、侵袭和 EMT。我们还发现,hsa_circ_0001681 沉默的过程限制了 EMT,这被 TWIST1 的增加所阻碍。此外,hsa_circ_0001681 作为 miRNA 海绵与 TWIST1 mRNA 结合完成与 miR-942-5p 的结合,从而通过靶向 miR-942-5p/TWIST1 信号通路下调 hsa_circ_0001681 抑制 EMT 和随后的 TC 细胞恶性表型。最后,体内研究表明,通过调节 miR-942-5p/TWIST1 信号通路抑制 EMT,降低 hsa_circ_0001681 的表达可有效抑制肿瘤的生长。总的来说,沉默 hsa_circ_0001681 通过作为 miR-942-5p 海绵来抑制 EMT,从而显著抑制 TC 进展,从而促进 TWIST1 的表达。我们的数据为 hsa_circ_0001681 作为 TC 潜在治疗靶点提供了可靠的证据。