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环状 RNA 结合蛋白 LINPEP 通过调控 miR-876-3p/WNT5A 轴促进卵巢癌细胞的进展。

CircLNPEP promotes the progression of ovarian cancer through regulating miR-876-3p/ WNT5A axis.

机构信息

Department of Gynaecology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou City, China.

出版信息

Cell Cycle. 2021 Oct;20(19):2021-2039. doi: 10.1080/15384101.2021.1965723. Epub 2021 Aug 31.

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 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

环形 RNA LNPEP 已被证明可促进肝细胞癌的发展,但它在卵巢癌 (OC) 中的功能尚不清楚。Kaplan-Meier 法分析 OC 患者中 circLNPEP 表达的临床意义。用放线菌素 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) 的相关性,并通过双荧光素酶报告基因实验证实。通过 qRT-PCR 和 Western blot 测定 circLNPEP、miR-876-3p 和 WNT5A 的表达。通过细胞功能实验和挽救实验确定 circLNPEP/miR-876-3p/WNT5A 轴对细胞活力、增殖、迁移和侵袭以及血管生成的影响。构建异种移植肿瘤小鼠进行验证。通过 qRT-PCR、Western blot 和免疫组化测定凋亡、上皮间质转化 (EMT) 相关基因和 CD34 的表达。OC 中高水平的 circLNPEP 与预后不良相关。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 靶向并负相关与 circLNPEP 和 WNT5A。Sh-circLNPEP 抑制细胞活力、增殖、迁移、侵袭、血管生成和 EMT,但促进凋亡,这与其对 EMT 和凋亡相关基因、WNT5A 和 CD34 的表达调节有关。sh-circLNPEP 的调节作用可以被 miR-876-3p 抑制剂逆转,而 miR-876-3p 抑制剂的调节作用可以被 sh-WNT5A 逆转。circLNPEP 通过调节 miR-876-3p/WNT5A 轴促进癌细胞增殖、EMT 和血管生成,抑制凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbcc/8565828/0c9ee4cfdb70/KCCY_A_1965723_F0001_OC.jpg

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