CircSCNN1A 通过 miR-590-5p 降低 CLDN8 的表达，从而抑制肾细胞癌细胞的增殖、迁移和侵袭。

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-05-19 DOI:10.1002/dvg.23599

Tingting Guo, Wanjuan Xiong, Chong Liu, Li Zhu, Ling Xie

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引用次数: 0

摘要

背景：越来越多的证据表明，环状 RNA（circRNA）在肾细胞癌（RCC）的进展过程中发挥着调控作用。然而，circSCNN1A在RCC进展中的确切功能和内在机制仍不清楚：方法：采用实时定量聚合酶链反应和 Western 印迹分析法检测 circSCNN1A、microRNA-590-5p（miR-590-5p）、claudin 8（CLDN8）、细胞周期蛋白 D1、基质金属蛋白 2（MMP2）、MMP9、E-钙粘连蛋白、N-钙粘连蛋白和波形蛋白的表达水平。免疫组化法分析了 CLDN8 的阳性表达率。通过细胞集落形成、5-乙炔基-2'-脱氧尿苷和 DNA 含量定量检测来研究细胞增殖。细胞迁移和侵袭通过伤口愈合和跨孔侵袭试验进行评估。通过双荧光素酶报告实验、RNA 免疫沉淀实验和 RNA 牵引实验确定了 circSCNN1A、miR-590-5p 和 CLDN8 之间的相互作用。通过异种移植小鼠模型试验验证了 circSCNN1A 对体内肿瘤形成的影响：结果：在 RCC 组织和细胞中，CircSCNN1A 和 CLDN8 的表达均显著下调，而 miR-590-5p 则上调。CircSCNN1A的过表达抑制了RCC细胞的增殖、迁移和侵袭，同时降低了细胞周期蛋白D1、MMP2、MMP9、N-粘连蛋白和波形蛋白的表达，增加了E-粘连蛋白的表达。CircSCNN1A作为miR-590-5p海绵，通过与miR-590-5p结合来调控RCC细胞的进程。miR-590-5p的靶基因CLDN8参与了miR-590-5p对RCC细胞生物学行为的调控。此外，circSCNN1A通过与miR-590-5p相互作用诱导CLDN8的产生。此外，circSCNN1A还抑制了体内肿瘤的形成：结论：circSCNN1A通过调节miR-590-5p/CLDN8通路抑制了RCC细胞的增殖、迁移和侵袭。

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CircSCNN1A inhibits the proliferation, migration and invasion of renal cell carcinoma cells by decreasing CLDN8 expression through miR-590-5p

Background

Increasing evidence suggests that circular RNA (circRNA) plays a regulatory role in the progression of renal cell carcinoma (RCC). However, the precise function and underlying mechanism of circSCNN1A in RCC progression still remain unclear.

Methods

The expression levels of circSCNN1A, microRNA-590-5p (miR-590-5p), claudin 8 (CLDN8), cyclin D1, matrix metalloprotein 2 (MMP2), MMP9, E-cadherin, N-cadherin and vimentin were detected by a quantitative real-time polymerase chain reaction and Western blotting analysis. Immunohistochemistry assay was performed to analyze the positive expression rate of CLDN8. Cell proliferation was investigated by cell colony formation, 5-Ethynyl-2′-deoxyuridine and DNA content quantitation assays. Cell migration and invasion were assessed by wound-healing and transwell invasion assays. Interactions among circSCNN1A, miR-590-5p and CLDN8 were identified by dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA pull-down assay. Xenograft mouse model assay was conducted to verify the effect of circSCNN1A on tumor formation in vivo.

Results

CircSCNN1A and CLDN8 expression were significantly downregulated, while miR-590-5p was upregulated in both RCC tissues and cells. CircSCNN1A overexpression inhibited RCC cell proliferation, migration and invasion, accompanied by decreases of cyclin D1, MMP2, MMP9, N-cadherin and vimentin expression and an increase of E-cadherin expression. CircSCNN1A acted as a miR-590-5p sponge and regulated RCC cell processes by binding to miR-590-5p. CLDN8, a target gene of miR-590-5p, was involved in the regulation of the biological behaviors of RCC cells by miR-590-5p. In addition, circSCNN1A induced CLDN8 production by interacting with miR-590-5p. Further, circSCNN1A suppressed tumor formation in vivo.

Conclusion

CircSCNN1A inhibited RCC cell proliferation, migration and invasion by regulating the miR-590-5p/CLDN8 pathway.

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