长链非编码 RNA ARAP1-AS1 通过调控 miR-149-3p 和 POU2F2 促进宫颈癌的进展。

Long Non-Coding RNA ARAP1-AS1 Facilitates the Progression of Cervical Cancer by Regulating miR-149-3p and POU2F2.

机构信息

Department of Obstetrics and Gynecology, Liyang People's Hospital, Liyang, China.

Department of Obstetrics and Gynecology, The First People's Hospital of Changzhou (The Third Affiliated Hospital of Suzhou University), Changzhou, China.

出版信息

Pathobiology. 2021;88(4):301-312. doi: 10.1159/000507830. Epub 2021 May 7.

DOI:10.1159/000507830

PMID:33965958

Abstract

BACKGROUND

Emerging research has demonstrated that long non-coding RNAs (lncRNAs) attach great importance to the progression of cervical cancer (CC). LncRNA ARAP1-AS1 was involved in the development of several cancers; however, its role in CC is far from being elucidated.

METHODS

Real-time PCR (RT-PCR) was employed to detect ARAP1-AS1 and miR-149-3p expression in CC samples. CC cell lines (HeLa and C33A cells) were regarded as the cell models. The biological effect of ARAP1-AS1 on cancer cells was measured using CCK-8 assay, colony formation assay, flow cytometry, Transwell assay and wound healing assay in vitro, and subcutaneous xenotransplanted tumor model and tail vein injection model in vivo. Furthermore, interactions between ARAP1-AS1 and miR-149-3p, miR-149-3p and POU class 2 homeobox 2 (POU2F2) were determined by bioinformatics analysis, qRT-PCR, Western blot, luciferase reporter and RNA immunoprecipitation assay, respectively.

RESULTS

The expression of ARAP1-AS1 was enhanced in CC samples, while miR-149-3p was markedly suppressed. Additionally, ARAP1-AS1 overexpression enhanced the viability, migration, and invasion of CC cells. ARAP1-AS1 downregulated miR-149-3p via sponging it. ARAP1-AS1 and miR-149-3p exhibited a negative correlation in CC samples. On the other hand, ARAP1-AS1 enhanced the expression of POU2F2, which was validated as a target gene of miR-149-3p.

CONCLUSION

ARAP1-AS1 was abnormally upregulated in CC tissues and indirectly modulated the POU2F2 expression via reducing miR-149-3p expression. Our study identified a novel axis, ARAP1-AS1/miR-149-3p/POU2F2, in CC tumorigenesis.

摘要

背景

新兴研究表明，长链非编码 RNA（lncRNA）对宫颈癌（CC）的进展具有重要意义。LncRNA ARAP1-AS1 参与了几种癌症的发展；然而，其在 CC 中的作用仍有待阐明。

方法

采用实时 PCR（RT-PCR）检测 CC 样本中 ARAP1-AS1 和 miR-149-3p 的表达。HeLa 和 C33A 细胞系被视为细胞模型。体外采用 CCK-8 测定法、集落形成测定法、流式细胞术、Transwell 测定法和划痕愈合测定法，以及体内皮下异种移植肿瘤模型和尾静脉注射模型，检测 ARAP1-AS1 对癌细胞的生物学效应。此外，通过生物信息学分析、qRT-PCR、Western blot、荧光素酶报告基因和 RNA 免疫沉淀测定法分别确定 ARAP1-AS1 和 miR-149-3p、miR-149-3p 和 POU 类 2 同源盒 2（POU2F2）之间的相互作用。

结果

CC 样本中 ARAP1-AS1 的表达增强，而 miR-149-3p 的表达明显受到抑制。此外，ARAP1-AS1 过表达增强了 CC 细胞的活力、迁移和侵袭。ARAP1-AS1 通过海绵作用下调 miR-149-3p。ARAP1-AS1 和 miR-149-3p 在 CC 样本中呈负相关。另一方面，ARAP1-AS1 增强了 POU2F2 的表达，这被验证为 miR-149-3p 的靶基因。