Yi Mo, Lv Samei, He Yu, Wu Youwei, Zhang Jian, Chen Qian
Department of Gastroenterology, Shaanxi Provincial People's Hospital, Xi'an, China.
Environ Toxicol. 2023 Aug;38(8):2031-2040. doi: 10.1002/tox.23831. Epub 2023 May 25.
Circular RNA (circRNA) is considered an important molecular marker for the early diagnosis of tumors and a potential therapeutic target. Herein, we investigated the role and regulatory mechanism of circKDM1B in hepatocellular carcinoma (HCC).
The expression of circKDM1B, miR-1322 and Protein regulator of cytokinesis 1 (PRC1) mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell counting kit-8 (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) staining assays were performed to assess cell proliferation activity. Cell migration and invasion were detected by wound-healing scratch and transwell assay. Flow cytometry was used to analyze cell apoptosis. The protein levels of PCNA, MMP9, C-caspase3 and PRC1 were examined using western blot. The binding of circKDM1B and miR-1322 was verified by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) and RNA pull down assay.
CircKDM1B was overexpressed in HCC tissues and cells, and its overexpression was related to tumor stage and poor prognosis of HCC patients. Functionally, knockdown of circKDM1B suppressed proliferation, migration, invasion and promoted apoptosis of HCC cells. Mechanistically, circKDM1B functioned as ceRNA of miR-1322 to upregulate PRC1 in HCC cells. Overexpression of miR-1322 inhibited proliferation, migration, invasion and facilitated apoptosis of HCC cells, which was partly reversed by PRC1 overexpression. CircKDM1B knockdown impeded HCC tumor growth in vivo.
CircKDM1B played a critical role in HCC progression by regulating cell proliferation, migration, invasion and apoptosis. CircKDM1B/miR-1322/PRC1 axis might be a novel therapeutic target of HCC patients.
环状RNA（circRNA）被认为是肿瘤早期诊断的重要分子标志物和潜在的治疗靶点。在此，我们研究了circKDM1B在肝细胞癌（HCC）中的作用及调控机制。
采用定量实时聚合酶链反应（qRT-PCR）检测circKDM1B、miR-1322和胞质分裂蛋白调节剂1（PRC1）mRNA的表达。进行细胞计数试剂盒-8（CCK8）和5-乙炔基-2'-脱氧尿苷（EdU）染色试验以评估细胞增殖活性。通过伤口愈合划痕试验和Transwell试验检测细胞迁移和侵袭。采用流式细胞术分析细胞凋亡。使用蛋白质免疫印迹法检测增殖细胞核抗原（PCNA）、基质金属蛋白酶9（MMP9）、半胱天冬酶3（C-caspase3）和PRC1的蛋白水平。通过双荧光素酶报告基因试验、RNA免疫沉淀（RIP）和RNA下拉试验验证circKDM1B与miR-1322的结合。
CircKDM1B在HCC组织和细胞中高表达，其高表达与HCC患者的肿瘤分期和不良预后相关。在功能上，敲低circKDM1B可抑制HCC细胞的增殖、迁移、侵袭并促进其凋亡。机制上，circKDM1B作为miR-1322的竞争性内源RNA（ceRNA）在HCC细胞中上调PRC1。miR-1322的过表达抑制HCC细胞的增殖、迁移、侵袭并促进其凋亡，而PRC1的过表达可部分逆转这一作用。敲低circKDM1B可抑制体内HCC肿瘤生长。
CircKDM1B通过调节细胞增殖、迁移、侵袭和凋亡在HCC进展中起关键作用。CircKDM1B/miR-1322/PRC1轴可能是HCC患者的新型治疗靶点。
