PAX8-AS1/microRNA-25–3p/LATS2 regulates malignant progression of ovarian cancer via Hippo signaling

IF 1.5 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis Pub Date : 2024-04-21 DOI:10.1016/j.mrfmmm.2024.111858

Gang Liu, Jing Tian

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引用次数: 0

Abstract

Background

Ovarian cancer (OC) is a frequent malignancy of the female reproductive system. Recently, the aberrant expression of numerous lncRNAs has been confirmed as a key factor for cancer development. The regulatory role of PAX8-AS1 in some cancers has been investigated, but its role in OC progression remains unclear. This study focuses on the role and molecular mechanism of PAX8-AS1 in the malignant progression of OC.

Methods

Bioinformatics means were adopted to analyze the expression of PAX8-AS1, microRNA-25–3p, and LATS2 in OC tissues and the binding sites between the three. qRT-PCR was employed to determine the expression of these genes in OC cells. CCK-8, colony formation, scratch healing, and Transwell assays were used to see cell viability, proliferation, migration, and invasion, respectively. Fluorescence in situ Hybridization was performed to probe the subcellular localization of PAX8-AS1. Western blot was applied to evaluate the expression and phosphorylation levels of YAP and TAZ, and an immunofluorescence assay was used to detect the translocation of them. Dual luciferase assay was applied to validate the binding relationship between PAX8-AS1 and microRNA-25–3p, as well as between microRNA-25–3p and LATS2.

Results

PAX8-AS1 and LATS2 were lowly expressed. MicroRNA-25–3p was highly expressed in OC. PAX8-AS1 was expressed in cytoplasm and regulated LATS2 expression by sponging microRNA-25–3p. Overexpressing PAX8-AS1 can suppress the malignant behaviors of OC cells, whereas treatment with microRNA-mimic can reverse these results. In addition, the phosphorylation levels of YAP and TAZ increased upon oe-LATS2 treatment, and oe-LATS2 could promote YAP and TAZ translocate from the nucleus to cytoplasm. Rescue experiments demonstrated that sh-PAX8-AS1 fostered malignant progression of OC, which was reversed by simultaneous oe-LATS2.

Conclusion

In summary, PAX8-AS1/microRNA-25–3p/LATS2 regulated the malignant progression of OC through Hippo signaling, which suggested that PAX8-AS1/microRNA-25–3p/LATS2 axis may be a novel target for OC treatment.

查看原文本刊更多论文

PAX8-AS1/microRNA-25-3p/LATS2通过Hippo信号调控卵巢癌的恶性进展。

背景：卵巢癌（OC）是女性生殖系统中一种常见的恶性肿瘤。最近，许多 lncRNAs 的异常表达已被证实是癌症发展的一个关键因素。PAX8-AS1在一些癌症中的调控作用已被研究，但其在OC进展中的作用仍不清楚。本研究主要探讨PAX8-AS1在OC恶性进展中的作用及分子机制：方法：采用生物信息学手段分析 PAX8-AS1、microRNA-25-3p 和 LATS2 在 OC 组织中的表达及三者之间的结合位点。CCK-8、集落形成、划痕愈合和 Transwell 试验分别用于检测细胞活力、增殖、迁移和侵袭。荧光原位杂交检测 PAX8-AS1 的亚细胞定位。用 Western 印迹法评估 YAP 和 TAZ 的表达和磷酸化水平，用免疫荧光法检测它们的转位。应用双荧光素酶试验验证了PAX8-AS1与microRNA-25-3p以及microRNA-25-3p与LATS2之间的结合关系：结果：PAX8-AS1和LATS2低表达。结果：PAX8-AS1 和 LATS2 的表达量较低，而 microRNA-25-3p 在 OC 中的表达量较高。PAX8-AS1 在细胞质中表达，并通过海绵状 microRNA-25-3p 调节 LATS2 的表达。过表达 PAX8-AS1 能抑制 OC 细胞的恶性行为，而用 microRNA 模拟物处理则能逆转这些结果。此外，oe-LATS2处理后，YAP和TAZ的磷酸化水平升高，oe-LATS2能促进YAP和TAZ从细胞核转移到细胞质。拯救实验表明，sh-PAX8-AS1可促进OC的恶性进展，而同时oe-LATS2可逆转恶性进展：综上所述，PAX8-AS1/microRNA-25-3p/LATS2通过Hippo信号调控OC的恶性进展，这表明PAX8-AS1/microRNA-25-3p/LATS2轴可能是治疗OC的新靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis 生物-毒理学

CiteScore

4.90

自引率

0.00%

发文量

审稿时长

51 days

期刊介绍： Mutation Research (MR) provides a platform for publishing all aspects of DNA mutations and epimutations, from basic evolutionary aspects to translational applications in genetic and epigenetic diagnostics and therapy. Mutations are defined as all possible alterations in DNA sequence and sequence organization, from point mutations to genome structural variation, chromosomal aberrations and aneuploidy. Epimutations are defined as alterations in the epigenome, i.e., changes in DNA methylation, histone modification and small regulatory RNAs. MR publishes articles in the following areas: Of special interest are basic mechanisms through which DNA damage and mutations impact development and differentiation, stem cell biology and cell fate in general, including various forms of cell death and cellular senescence. The study of genome instability in human molecular epidemiology and in relation to complex phenotypes, such as human disease, is considered a growing area of importance. Mechanisms of (epi)mutation induction, for example, during DNA repair, replication or recombination; novel methods of (epi)mutation detection, with a focus on ultra-high-throughput sequencing. Landscape of somatic mutations and epimutations in cancer and aging. Role of de novo mutations in human disease and aging; mutations in population genomics. Interactions between mutations and epimutations. The role of epimutations in chromatin structure and function. Mitochondrial DNA mutations and their consequences in terms of human disease and aging. Novel ways to generate mutations and epimutations in cell lines and animal models.