Department of Clinical Laboratory, Affiliated Neijiang Second People's Hospital of Southwest Medical University, Neijiang, P.R, China.
Department of Clinical Laboratory, The First Affiliated Hospital of Southwest Medical University, China Neijiang.
Bioengineered. 2022 Apr;13(4):10578-10593. doi: 10.1080/21655979.2022.2063562.
Recent studies have shown that circRNAs can act as oncogenic factors or tumor suppressors by sponging microRNAs (miRNAs). The upregulation of circ_0023984 was reported in esophageal squamous cell carcinoma (ESCC). However, its functional role in ESCC remain unclear. In the present study, circ_0023984 expression in ESCC cells and tissues were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB). Subcellular fraction experiment was performed to determine relative nuclear-cytoplasmic localization. The loss-of-function effects of circ_0023984 in ESCC cell lines were investigated by shRNA-mediated knockdown. Functional assays including cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EDU) incorporation, colony formation and Transwell migration assays were conducted to assess the malignant phenotype. The interaction between the two molecules was analyzed by RNA pull-down, luciferase reporter assay and RNA immunoprecipitation (RIP). The subcutaneous tumor model in nude mice was used to assess the role of circ-0023984 in tumorigenesis. We found that ESCC patients with high circ_0023984 expression was associated with a poor prognosis. The knockdown of circ_0023984 suppressed cell growth, invasion, and migration in ESCC cells. Circ_0023984 interacted with miR-134-5p and inhibited its activity, which promoted the expression of CST4 (Cystatin-S). Circ_0023984 also regulated tumorigenesis in a CST4-dependent manner. Together, our study indicates that the oncogenic role of Circ_0023984 is mediated by miR-134-5p/CST4 Axis in ESCC, which could serve as potential targets for future therapeutic strategies.
最近的研究表明，circRNAs 可以通过海绵吸附 microRNAs（miRNAs）来作为致癌因子或肿瘤抑制因子。circ_0023984 在食管鳞状细胞癌（ESCC）中被报道上调。然而，其在 ESCC 中的功能作用仍不清楚。在本研究中，通过实时定量聚合酶链反应（qRT-PCR）和 Western blot（WB）分析了 ESCC 细胞和组织中的 circ_0023984 表达。进行亚细胞分离实验以确定相对核-质定位。通过 shRNA 介导的敲低研究了 circ_0023984 在 ESCC 细胞系中的功能缺失效应。通过细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EDU）掺入、集落形成和 Transwell 迁移实验进行功能测定，以评估恶性表型。通过 RNA 下拉、荧光素酶报告基因测定和 RNA 免疫沉淀（RIP）分析两个分子之间的相互作用。使用裸鼠皮下肿瘤模型评估 circ-0023984 在肿瘤发生中的作用。我们发现，ESCC 患者中高表达 circ_0023984 与预后不良相关。circ_0023984 的敲低抑制了 ESCC 细胞的生长、侵袭和迁移。circ_0023984 与 miR-134-5p 相互作用并抑制其活性，从而促进 CST4（胱抑素-S）的表达。circ_0023984 还以 CST4 依赖的方式调节肿瘤发生。总之，我们的研究表明，Circ_0023984 的致癌作用是通过 miR-134-5p/CST4 轴在 ESCC 中介导的，这可能成为未来治疗策略的潜在靶点。
