CircRAB3B suppresses proliferation, motility, cell cycle progression and promotes the apoptosis of IL-22-induced keratinocytes depending on the regulation of miR-1228-3p/PTEN axis in psoriasis,Autoimmunity

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CircRAB3B suppresses proliferation, motility, cell cycle progression and promotes the apoptosis of IL-22-induced keratinocytes depending on the regulation of miR-1228-3p/PTEN axis in psoriasis
Autoimmunity ( IF 3.5 ) Pub Date : 2021-06-07 , DOI: 10.1080/08916934.2021.1934825
Jiajing Lu _{1,

2} , Xin Xu _{1,

2} , Ying Li _{1,

2} , Ning Yu _{1,

2} , Yangfeng Ding _{1,

2} , Yuling Shi _{1,

2}

Affiliation

Abstract

Introduction

Psoriasis is an immune-related chronic skin disease, and interleukin-22 (IL-22) is involved in psoriasis pathogenesis through promoting proliferation and migration abilities of keratinocytes. Here, we analysed the role of circular RNA (circRNA) RAB3B, member RAS oncogene family (circRAB3B) in regulating the phenotypes of IL-22-induced HaCaT cells.

Methods

RT-qPCR was implemented to assess RNA abundance. Western blot assay was adopted to assess protein abundance. Cell proliferation capacity was examined by cell counting kit-8 (CCK8) assay and 5-ethynyl-2′-deoxyuridine (Edu) assay. Cell motility was assessed by transwell assays and wound healing assay. Flow cytometric analysis was utilized to evaluate cell cycle progression and apoptosis. The intermolecular binding relations were tested via dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. CircRAB3B expression was reduced in psoriatic cutaneous specimens and IL-22-treated HaCaT cells.

Results

CircRAB3B overexpression hampered the proliferation, motility, and cell cycle progression and elevated the apoptotic rate of IL-22-treated HaCaT cells, and circRAB3B silencing exhibited opposite effects in IL-22-induced HaCaT cells. CircRAB3B acted as microRNA-1228-3p (miR-1228-3p) sponge in HaCaT cells, and miR-1228-3p overexpression largely overturned circRAB3B overexpression-induced effects in HaCaT cells. MiR-1228-3p interacted with phosphatase and tensin homolog (PTEN), and circRAB3B sponged miR-1228-3p to induce PTEN level. MiR-1228-3p accumulation-mediated effects were partly alleviated by PTEN overexpression in HaCaT cells upon IL-22 treatment.

Conclusions

CircRAB3B suppressed psoriasis progression partly through down-regulating miR-1228-3p and up-regulating PTEN.

中文翻译：

CircRAB3B 通过调节银屑病中 miR-1228-3p/PTEN 轴抑制增殖、运动、细胞周期进程并促进 IL-22 诱导的角质形成细胞凋亡

摘要

介绍

银屑病是一种免疫相关的慢性皮肤病，白细胞介素22（IL-22）通过促进角质形成细胞的增殖和迁移能力参与银屑病的发病机制。在这里，我们分析了环状 RNA (circRNA) RAB3B，RAS 癌基因家族 (circRAB3B) 成员在调节 IL-22 诱导的 HaCaT 细胞表型中的作用。

方法

实施 RT-qPCR 以评估 RNA 丰度。采用蛋白质印迹法评估蛋白质丰度。通过细胞计数试剂盒-8 (CCK8) 测定和 5-乙炔基-2'-脱氧尿苷 (Edu) 测定检查细胞增殖能力。通过transwell测定和伤口愈合测定评估细胞运动性。流式细胞术分析用于评估细胞周期进程和细胞凋亡。通过双荧光素酶报告基因测定和 RNA 免疫沉淀 (RIP) 测定测试分子间结合关系。银屑病皮肤标本和经 IL-22 处理的 HaCaT 细胞中 CircRAB3B 表达降低。

结果

CircRAB3B 过表达阻碍了 IL-22 处理的 HaCaT 细胞的增殖、运动和细胞周期进程，并提高了凋亡率，而 circRAB3B 沉默在 IL-22 诱导的 HaCaT 细胞中表现出相反的作用。CircRAB3B 在 HaCaT 细胞中充当 microRNA-1228-3p (miR-1228-3p) 海绵，miR-1228-3p 过表达在很大程度上推翻了 circRAB3B 过表达诱导的 HaCaT 细胞效应。MiR-1228-3p 与磷酸酶和张力蛋白同源物 (PTEN) 相互作用，circRAB3B 海绵化 miR-1228-3p 以诱导 PTEN 水平。在 IL-22 处理后，HaCaT 细胞中的 PTEN 过表达部分减轻了 MiR-1228-3p 积累介导的作用。