ABSTRACT

CircRNA LNPEP has been shown to promote the development of hepatocellular carcinoma, but its function in ovarian cancer (OC) remains unclear. The Kaplan–Meier method was used to analyze the clinical significance of circLNPEP expression in OC patients. The stability of circLNPEP was detected by actinomycin D and RNase R treatment. The correlations between miR-876-3p and two genes (circLNPEP and WNT5A) were predicted by bioinformatics analysis and confirmed by dual-luciferase reporter assay. Expressions of circLNPEP, miR-876-3p, and WNT5A were determined by qRT-PCR and western blot. The effect of circLNPEP/miR-876-3p/WNT5A axis on viability, proliferation, migration, and invasion, and angiogenesis of cells was determined by cell function experiment and rescue experiment. Xenograft tumor mice were constructed for in vivo verification. Expressions of apoptosis, epithelial mesenchymal transition (EMT)-related genes, and CD34 were determined by qRT-PCR, western blot and immunohistochemistry. High level of circLNPEP was related to poor prognosis in OC. CircLNPEP was highly expressed in OC tissues and cell lines, mainly distributed in the cytoplasm, while miR-876-3p was the opposite. MiR-876-3p targeted and negatively correlated with circLNPEP and WNT5A. Sh-circLNPEP repressed cell viability, proliferation, migration, invasion, angiogenesis, and EMT but promoted apoptosis, which were related to its regulation of expression of EMT- and apoptosis-related genes, WNT5A, and CD34. The regulatory effect of sh-circLNPEP can be reversed by miR-876-3p inhibitor, and that of miR-876-3p inhibitor can be reversed by sh-WNT5A. CircLNPEP promoted cancer cell proliferation, EMT and angiogenesis, and inhibited apoptosis by regulating miR-876-3p/WNT5A axis.

摘要

CircRNA LNPEP 已被证明可促进肝细胞癌的发展，但其在卵巢癌 (OC) 中的作用仍不清楚。Kaplan-Meier 法用于分析 oc 患者中 circLNPEP 表达的临床意义。通过放线菌素D和RNase R处理检测circLNPEP的稳定性。通过生物信息学分析预测 miR-876-3p 与两个基因（circLNPEP 和 WNT5A）之间的相关性，并通过双荧光素酶报告基因分析证实。通过 qRT-PCR 和蛋白质印迹测定 circLNPEP、miR-876-3p 和 WNT5A 的表达。通过细胞功能实验和拯救实验确定circLNPEP/miR-876-3p/WNT5A轴对细胞活力、增殖、迁移、侵袭和血管生成的影响。为体内构建异种移植肿瘤小鼠确认。通过 qRT-PCR、蛋白质印迹和免疫组织化学测定细胞凋亡、上皮间质转化 (EMT) 相关基因和 CD34 的表达。高水平的 circLNPEP 与 OC 的不良预后有关。CircLNPEP在OC组织和细胞系中高表达，主要分布在细胞质中，而miR-876-3p则相反。MiR-876-3p 靶向并与 circLNPEP 和 WNT5A 呈负相关。Sh-circLNPEP 抑制细胞活力、增殖、迁移、侵袭、血管生成和 EMT，但促进细胞凋亡，这与其调节 EMT 和凋亡相关基因 WNT5A 和 CD34 的表达有关。sh-circLNPEP的调节作用可以被miR-876-3p inhibitor逆转，miR-876-3p inhibitor的调节作用可以被sh-WNT5A逆转。CircLNPEP 促进癌细胞增殖，