UCA1通过调控miR-582-5p/BRCC3在胰腺癌中发挥致癌作用。

UCA1 executes an oncogenic role in pancreatic cancer by regulating miR-582-5p/BRCC3.

作者信息

Hu Xiaole, Wu Jiahao, Xu Jianwei

机构信息

Department of First Operating Room, Qilu Hospital of Shandong University, Jinan, China.

Department of Pancreatic Surgery, General Surgery, Qilu Hospital, Shandong University, Jinan, China.

出版信息

Front Oncol. 2023 Jul 25;13:1133200. doi: 10.3389/fonc.2023.1133200. eCollection 2023.

DOI:10.3389/fonc.2023.1133200

PMID:37564930

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10411552/

Abstract

BACKGROUND

As a fatal disease, the mechanism of pancreatic cancer is unclear. Urothelial carcinoma antigen 1(UCA1), a long noncoding RNA (lncRNA) that was first reported in bladder cancer, acts as an oncogene. However, the regulatory role and mechanism of UCA1 in pancreatic cancer remain unknown. This study aims to investigate the expression level and prognostic value of UCA1 in pancreatic cancer tissues, the effects and mechanism of UCA1 in regulating cell proliferation, apoptosis and metastasis.

METHODS

UCA1 expression levels in tissues were detected by in situ hybridization (ISH) and the prognostic value was evaluated by univariate and multivariate survival analysis. For experiments, proliferation was evaluated by a cell count kit assay, Edu experiments, and a clone formation assay. Apoptosis was evaluated by fluorescence-activated cell sorting flow-cytometry. Cell migration and invasion capacities were detected by wound healing and transwell assays. Western blots were performed to detect apoptotic associated molecules and epithelial-mesenchymal transition (EMT) markers. For the experiment, subcutaneous transplantation models of pancreatic cancer in nude mice were established to observe the tumor growth. The regulatory mechanism of UCA1 was explored by proteomics, bioinformatic analysis, luciferase reporter assays, and rescue experiments.

RESULTS

ISH staining revealed that UCA1 levels between cancer tissues (n=94) and tumor-adjacent tissues (n=73) did not show significant differences. Survival analysis indicated that high expression of UCA1 was an unfavorable prognosis factor for pancreatic cancer. Downregulation of UCA1 by siRNA significantly inhibited cell proliferation, decreased the capacities of cell migration and invasion, induced cell apoptosis, and inhibited EMT. Furthermore, we demonstrated that UCA1 positively regulated the expression of BRCC3 by inhibiting miR-582-5p. Rescue experiments indicated that either inhibiting the expression of miR-582-5p or enhancing expression of BRCC3 could partly attenuate the antitumor effects of downregulation of UCA1.

CONCLUSION

UCA1 acted as an oncogene in pancreatic cancer by partly regulating miR-582-5p/BRCC3, which could be a new therapeutic target for pancreatic cancer.

摘要

背景

胰腺癌作为一种致命疾病，其发病机制尚不清楚。尿路上皮癌抗原1（UCA1）是一种长链非编码RNA（lncRNA），最初在膀胱癌中被报道，具有癌基因作用。然而，UCA1在胰腺癌中的调控作用和机制仍不清楚。本研究旨在探讨UCA1在胰腺癌组织中的表达水平及预后价值，以及UCA1在调节细胞增殖、凋亡和转移中的作用及机制。

方法

采用原位杂交（ISH）检测组织中UCA1表达水平，并通过单因素和多因素生存分析评估其预后价值。体外实验中，通过细胞计数试剂盒检测、Edu实验和克隆形成实验评估细胞增殖。通过荧光激活细胞分选流式细胞术评估细胞凋亡。通过伤口愈合实验和Transwell实验检测细胞迁移和侵袭能力。采用蛋白质印迹法检测凋亡相关分子和上皮-间质转化（EMT）标志物。体内实验中，建立裸鼠胰腺癌皮下移植模型观察肿瘤生长。通过蛋白质组学、生物信息学分析、荧光素酶报告基因实验和挽救实验探索UCA1的调控机制。

结果

ISH染色显示，癌组织（n = 94）和癌旁组织（n = 73）中UCA1水平无显著差异。生存分析表明，UCA1高表达是胰腺癌的不良预后因素。siRNA下调UCA1可显著抑制细胞增殖，降低细胞迁移和侵袭能力，诱导细胞凋亡，并抑制EMT。此外，我们证明UCA1通过抑制miR-582-5p正向调节BRCC3的表达。挽救实验表明，抑制miR-582-5p表达或增强BRCC3表达均可部分减弱下调UCA1的抗肿瘤作用。