长链非编码RNA AGAP2-AS1与IGF2BP2相互作用，通过调节LRG1 mRNA稳定性促进膀胱癌进展。

LncRNA AGAP2-AS1 interacts with IGF2BP2 to promote bladder cancer progression via regulating LRG1 mRNA stability.

作者信息

Zhao Xu, Chen Jinbo, Zhang Chunyu, Xie Guoou, Othmane Belaydi, Kuang Xiaogen, Liu Bolong

机构信息

Department of Urology, Xiangya Hospital, Central South University, Changsha 410008, Hunan, PR China; Department of Urology, Guizhou Provincial People's Hospital, Guiyang 550001, Guizhou, PR China.

Department of Urology, Xiangya Hospital, Central South University, Changsha 410008, Hunan, PR China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, PR China.

出版信息

Cell Signal. 2023 Nov;111:110839. doi: 10.1016/j.cellsig.2023.110839. Epub 2023 Aug 2.

DOI:10.1016/j.cellsig.2023.110839

PMID:37541640

Abstract

BACKGROUND

The long non-coding RNA (lncRNA) AGAP2-AS1 was implicated in tumorigenesis, yet with unclear mechanism in the development of Bladder Cancer (BCa).

METHODS

We collected the clinicopathological features and tissue samples of 45 patients with BCa in Xiangya Hospital. Expressions of AGAP2-AS1 and LRG1 were detected by RT-qPCR in BCa tissues and normal tissues as well as in BCa cells. The roles of AGAP2-AS1 and LRG1 were investigated by CCK-8, colony formation assay, transwell assays and tube formation assay. The subcellular localization of AGAP2-AS1 was detected by Fluorescence in situ hybridization. Bioinformatics method, RNA immunoprecipitation, RNA pull-down assay and Actinomycin D test were used to predict and identify the relationships between AGAP2-AS1, LRG1 and IGF2BP2. Xenografted tumors were produced to explore the function of AGAP2-AS1 in BCa in vivo.

RESULTS

AGAP2-AS1 and LRG1 were highly upregulated in BCa. AGAP2-AS1 positively correlated with T stage, grade and vascular invasion, but negatively correlated with the survival of patients. Overexpressions of AGAP2-AS1 promoted proliferation, migration, invasion, tumor angiogenesis in vitro and tumor growth, metastasis in vivo, knockdown of AGAP2-AS1 exhibited the opposite effects. AGAP2-AS1 localized mainly in the cytoplasm. AGAP2-AS1 directly bound to IGF2BP2 protein to enhance LRG1 mRNA stability. Inhibition of BCa progression by AGAP2-AS1 knockdown may be reversed by LRG1 overexpression.

CONCLUSION

AGAP2-AS1 can promote BCa progression and metastasis by recruiting IGF2BP2 to stabilize LRG1.

摘要

背景

长链非编码RNA（lncRNA）AGAP2-AS1与肿瘤发生有关，但其在膀胱癌（BCa）发生发展中的机制尚不清楚。

方法

我们收集了湘雅医院45例BCa患者的临床病理特征和组织样本。采用RT-qPCR检测BCa组织、正常组织及BCa细胞中AGAP2-AS1和LRG1的表达。通过CCK-8、集落形成实验、Transwell实验和管腔形成实验研究AGAP2-AS1和LRG1的作用。采用荧光原位杂交检测AGAP2-AS1的亚细胞定位。运用生物信息学方法、RNA免疫沉淀、RNA下拉实验和放线菌素D实验预测并鉴定AGAP2-AS1、LRG1和IGF2BP2之间的关系。构建异种移植瘤以探讨AGAP2-AS1在体内BCa中的功能。

结果

AGAP2-AS1和LRG1在BCa中高度上调。AGAP2-AS1与T分期、分级和血管侵犯呈正相关，但与患者生存率呈负相关。AGAP2-AS1的过表达促进体外增殖、迁移、侵袭、肿瘤血管生成以及体内肿瘤生长和转移，敲低AGAP2-AS1则表现出相反的作用。AGAP2-AS1主要定位于细胞质。AGAP2-AS1直接与IGF2BP2蛋白结合以增强LRG1 mRNA的稳定性。LRG1的过表达可能会逆转敲低AGAP2-AS1对BCa进展的抑制作用。