RNF19A 通过调控 ILK 的泛素化并抑制 AKT/mTOR 信号通路来抑制膀胱癌的进展。

RNF19A inhibits bladder cancer progression by regulating ILK ubiquitination and inactivating the AKT/mTOR signalling pathway.

机构信息

Department of Urology, The First Affiliated Hospital of Yangtze University, The First people's Hospital of Jingzhou, Jingzhou, 434000, China.

Department of Urology, Renmin Hospital of Wuhan University, Wuhan, 430060, China.

出版信息

Biol Direct. 2024 Nov 6;19(1):102. doi: 10.1186/s13062-024-00562-2.

DOI:10.1186/s13062-024-00562-2

PMID:39508245

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11539788/

Abstract

BACKGROUND

The role of the RING finger protein superfamily in carcinogenesis has been widely studied, but one member of this family, RNF19A, has not yet been thoroughly explored in bladder cancer (BCa).

METHODS

The expression levels of RNF19A in BCa samples and cell lines were analysed through data mining of public resources and further experiments. BCa cells in which RNF19A was stably overexpressed or knocked down were generated through lentivirus infection. The effects of RNF19A on cell proliferation, migration, and invasion were explored by performing a series of in vitro experiments, including CCK-8, colony formation, wound healing, and Transwell invasion assays. Using bioinformatics methods and multiple experiments, including western blot, qRT‒PCR, immunoprecipitation, cycloheximide, ubiquitination, and rescue assays, the mechanism underlying the effect of RNF19A on the progression of BCa was investigated.

RESULTS

Here, we found that RNF19A expression was reduced in BCa samples and cell lines and that lower RNF19A expression predicted shorter overall survival of BCa patients. Functionally, forced expression of RNF19A suppressed BCa cell proliferation, migration, and invasion by inactivating the AKT/mTOR signalling pathway, whereas silencing RNF19A had the opposite effects. Mechanistically, RNF19A could directly interact with ILK and promote its ubiquitination and degradation. Rescue experiments revealed that forced ILK expression partially rescued the decreased phosphorylation of AKT, mTOR, and S6K1 caused by RNF19A overexpression and that the increased levels of the p-AKT, p-mTOR, and p-S6K1 proteins induced by RNF19A knockdown were eliminated after silencing ILK. Similarly, the effects of RNF19A overexpression or knockdown on the phenotypes of cell proliferation, migration, and invasion could also be restored by forced or decreased ILK expression.

CONCLUSIONS

RNF19A suppressed the proliferation, migration, and invasion abilities of BCa cells by regulating ILK ubiquitination and inactivating the AKT/mTOR signalling pathway. RNF19A might be a potential prognostic biomarker and promising therapeutic target for BCa.

摘要

背景

RING 指蛋白家族在致癌作用中的作用已得到广泛研究，但该家族的一个成员 RNF19A 在膀胱癌（BCa）中的研究尚未深入。

方法

通过公共资源的数据挖掘和进一步实验分析 RNF19A 在 BCa 样本和细胞系中的表达水平。通过慢病毒感染生成 RNF19A 稳定过表达或敲低的 BCa 细胞。通过一系列体外实验，包括 CCK-8、集落形成、划痕愈合和 Transwell 侵袭实验，研究 RNF19A 对细胞增殖、迁移和侵袭的影响。使用生物信息学方法和包括 Western blot、qRT-PCR、免疫沉淀、环己酰亚胺、泛素化和挽救实验在内的多个实验，研究了 RNF19A 对 BCa 进展影响的机制。

结果

在这里，我们发现 RNF19A 的表达在 BCa 样本和细胞系中降低，并且较低的 RNF19A 表达预示着 BCa 患者的总生存期较短。功能上，通过使 AKT/mTOR 信号通路失活，强制表达 RNF19A 抑制了 BCa 细胞的增殖、迁移和侵袭，而沉默 RNF19A 则产生相反的效果。从机制上讲，RNF19A 可以直接与 ILK 相互作用，并促进其泛素化和降解。挽救实验表明，强制表达 ILK 部分挽救了 RNF19A 过表达导致的 AKT、mTOR 和 S6K1 磷酸化水平降低，而 RNF19A 敲低引起的 p-AKT、p-mTOR 和 p-S6K1 蛋白水平升高在沉默 ILK 后消除。同样，RNF19A 过表达或敲低对细胞增殖、迁移和侵袭表型的影响也可以通过强制或降低 ILK 表达来恢复。