Chen Wenbin, Cen Shengren, Zhou Xumin, Yang Taowei, Wu Kaihui, Zou Libin, Luo Junqi, Li Chuanyin, Lv Daojun, Mao Xiangming
Department of Urology, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
Guangdong Key Laboratory of Urology, Department of Urology, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Front Cell Dev Biol. 2021 Jan 8;8:624764. doi: 10.3389/fcell.2020.624764. eCollection 2020.
CircRNAs recently have shown critical roles in tumor biology. However, their roles in prostate cancer (PCa) remains largely unclear.
CircRNA microarrays were performed in immortal prostate cell line RWPE1 and PCa cell lines as DU145, PC3, LNCaP, C4-2, and 22RV1. Combined with upregulated circRNAs in PCa tissues, circNOLC1 expression was validated in PCa cells and tissues qRT-PCR and FISH. Sanger sequencing, actinomycin D, gDNA, and cDNA, RNase R assays were used to assess the circular characteristics of circNOLC1. CCK-8, colony formation, transwell migration assays, and mice xenograft models were conducted to evaluate the functions of PCa cells after circNOLC1 knockdown and overexpression. RNA pulldown, luciferase reporter assay, FISH (fluorescence hybridization), and CHIP were utilized to illustrate the further mechanisms of circNOLC1.
Our research indicated that circNOLC1 was overexpressed in PCa cells and tissues, and circNOLC1 was more stable than linear NOLC1 mRNA. CircNOLC1 promoted PCa cells proliferation and migration and . Additionally, we found that circNOLC1 could upregulate PAQR4 expression by sponging miR-647, leading to the activation of PI3K/Akt pathway. Moreover, NF-kappaB was identified to bind to the NOLC1 promoter sites and upregulated both NOLC1 and circNOLC1 expression.
CircNOLC1, elevated by transcription factor NF-kappaB, promotes PCa progression a miR-647/PAQR4 axis, and circNOLC1 is a potential biomarker and target for PCa treatment.
环状RNA（circRNAs）最近在肿瘤生物学中显示出关键作用。然而，它们在前列腺癌（PCa）中的作用仍 largely不清楚。
在永生化前列腺细胞系RWPE1和PCa细胞系如DU145、PC3、LNCaP、C4 - 2和22RV1中进行circRNA微阵列分析。结合PCa组织中上调的circRNAs，通过qRT - PCR和FISH在PCa细胞和组织中验证circNOLC1的表达。使用桑格测序、放线菌素D、基因组DNA（gDNA）和互补DNA（cDNA）、RNase R分析来评估circNOLC1的环状特征。进行CCK - 8、集落形成、Transwell迁移分析和小鼠异种移植模型，以评估circNOLC1敲低和过表达后PCa细胞的功能。利用RNA下拉、荧光素酶报告基因分析、FISH（荧光杂交）和染色质免疫沉淀（CHIP）来说明circNOLC1的进一步机制。
我们的研究表明circNOLC1在PCa细胞和组织中过表达，并且circNOLC1比线性NOLC1 mRNA更稳定。circNOLC1促进PCa细胞增殖和迁移。此外，我们发现circNOLC1可以通过海绵吸附miR - 647上调PAQR4表达，导致PI3K/Akt途径激活。此外，核因子κB（NF - κB）被鉴定为与NOLC1启动子位点结合并上调NOLC1和circNOLC1的表达。
由转录因子NF - κB上调的circNOLC1通过miR - 647/PAQR4轴促进PCa进展，并且circNOLC1是PCa治疗的潜在生物标志物和靶点。
