Department of General Surgery, Wujin Hospital of Traditional Chinese Medicine, Changzhou, China.
Department of Hepatobiliary Surgery, Xinghua People's Hospital Affiliated Yangzhou University, Xinghua, China.
Sci Rep. 2024 Jul 23;14(1):16913. doi: 10.1038/s41598-024-67819-8.
Although circular RNAs (circRNA) have been demonstrated to modulate tumor initiation and progression, their roles in the proliferation of hepatocellular carcinoma (HCC) are still poorly understood. Based on the analysis of GEO data (GSE12174), hsa-circRNA-0015004 (circ-0015004) was screened and validated in 80 sets of HCC specimens. Subcellular fractionation analysis was designed to determine the cellular location of circ-0015004. Colony formation and cell counting kit-8 were performed to investigate the role of circ-0015004 in HCC. Dual-luciferase reporter gene assays, RNA immunoprecipitation and chromatin immunoprecipitation were employed to verify the interaction among circ-0015004, miR-330-3p and regulator of chromatin condensation 2 (RCC2). The expression level of circ-0015004 was significantly upregulated in HCC cell lines and HCC tissues. HCC patients with higher circ-0015004 levels displayed shorter overall survival, and higher tumor size and TNM stage. Moreover, knockdown of circ-0015004 significantly reduced HCC cell proliferation in vitro and inhibited the growth of HCC in nude mice. Mechanistic studies revealed that circ-0015004 could upregulate the expression of RCC2 by sponging miR-330-3p, thereby promoting HCC cell proliferation. Furthermore, we identified that Ying Yang 1 (YY1) could function as an important regulator of circ-0015004 transcription. This study systematically demonstrated the novel regulatory signaling of circ-0015004/miR-330-3p/RCC2 axis in promoting HCC progression, providing insight into HCC diagnosis and treatment from bench to clinic.
虽然环状 RNA(circRNA)已被证明可调节肿瘤的发生和发展,但它们在肝细胞癌(HCC)增殖中的作用仍知之甚少。基于 GEO 数据(GSE12174)的分析,筛选并验证了 80 组 HCC 标本中的 hsa-circRNA-0015004(circ-0015004)。设计亚细胞分馏分析以确定 circ-0015004 的细胞位置。进行集落形成和细胞计数试剂盒-8 实验以研究 circ-0015004 在 HCC 中的作用。双荧光素酶报告基因检测、RNA 免疫沉淀和染色质免疫沉淀实验用于验证 circ-0015004、miR-330-3p 和染色质凝聚调节因子 2(RCC2)之间的相互作用。circ-0015004 在 HCC 细胞系和 HCC 组织中的表达水平显著上调。circ-0015004 水平较高的 HCC 患者总生存期较短,肿瘤体积和 TNM 分期较高。此外,circ-0015004 的敲低显著降低了 HCC 细胞的体外增殖,并抑制了裸鼠中 HCC 的生长。机制研究表明,circ-0015004 可以通过海绵吸附 miR-330-3p 而上调 RCC2 的表达,从而促进 HCC 细胞增殖。此外,我们发现 Ying Yang 1(YY1)可以作为 circ-0015004 转录的重要调节因子。这项研究系统地证明了 circ-0015004/miR-330-3p/RCC2 轴在促进 HCC 进展中的新的调控信号,为从实验室到临床的 HCC 诊断和治疗提供了新的思路。