Department of Hepatobiliary Surgery, The Affiliated Hospital of Medical School, Ningbo University, Ningbo, Zhejiang 315020, P.R. China.
Int J Mol Med. 2020 Aug;46(2):729-739. doi: 10.3892/ijmm.2020.4632. Epub 2020 Jun 4.
Long non‑coding RNA (lncRNA) DGCR5 has been identified as a tumor suppressor in several types of cancer. However, its biological functions in pancreatic cancer (PaCa) have not yet been fully elucidated. The present study was designed to investigate the role of lncRNA DGCR5 in the regulation of PaCa cell apoptosis. For this purpose, lncRNA DGCR5, miR‑27a‑3p and Bcl‑2/adenovirus E1B‑19kDa‑interacting protein 3 (BNIP3) expression levels were examined by reverse transcription‑quantitative (RT‑qPCR) and western blot analysis, respectively. RNA pull‑down assay was used to verify DGCR5 as a target of miR‑27a‑3p and dual luciferase reporter assay was used to clarify whether miR‑27a‑3p targets the BNIP3 3' UTR. In addition, PaCa cell apoptosis was assessed by flow cytometry. Recombinant plasmids and cell transfection were performed to modulate the endogenous expression of related genes. Thereafter, the role of DGCR5 in PaCa was analyzed using a nude mouse model of PaCa. lncRNA DGCR5 was found to be downregulated in PaCa tissues and cells. DGCR5 functioned as a decoy of miR‑27a‑3p, and BNIP3 was negatively regulated by miR‑27a‑3p. Following the transfection of DGCR5 plasmid into PaCa cells, the expression of miR‑27a‑3p was downregulated, and this downregulation was reversed following transfection with miR‑27a‑3p mimic. In addition, DGCR5 regulated the BNIP3 and p38 MAPK pathways via miR‑27a‑3p and promoted PaCa cell apoptosis via the miR‑27a‑3p/BNIP3 pathway. The results of in vivo experiments also indicated the positive effects of DGCR5 on a nude mouse model of PaCa. On the whole, the findings of the present study indicate that lncRNA DGCR5 upregulates the BNIP3 and p38 MAPK pathways via miR‑27a‑3p to promote PaCa cell apoptosis, thereby attenuating PaCa development.
长链非编码 RNA (lncRNA) DGCR5 已被鉴定为多种类型癌症中的肿瘤抑制因子。然而，其在胰腺癌 (PaCa) 中的生物学功能尚未完全阐明。本研究旨在探讨 lncRNA DGCR5 在调节 PaCa 细胞凋亡中的作用。为此，通过逆转录-定量 (RT-qPCR) 和 Western blot 分析分别检测 lncRNA DGCR5、miR-27a-3p 和 Bcl-2/腺病毒 E1B-19kDa 相互作用蛋白 3 (BNIP3) 的表达水平。使用 RNA 下拉实验验证 DGCR5 是 miR-27a-3p 的靶标，使用双荧光素酶报告实验阐明 miR-27a-3p 是否靶向 BNIP3 3'UTR。此外，通过流式细胞术评估 PaCa 细胞凋亡。通过重组质粒和细胞转染来调节相关基因的内源性表达。之后，使用 PaCa 裸鼠模型分析 DGCR5 在 PaCa 中的作用。研究发现，lncRNA DGCR5 在 PaCa 组织和细胞中下调。DGCR5 作为 miR-27a-3p 的诱饵发挥作用，而 BNIP3 受 miR-27a-3p 的负调控。将 DGCR5 质粒转染入 PaCa 细胞后，miR-27a-3p 的表达下调，而转染 miR-27a-3p 模拟物后则逆转了这一下调。此外，DGCR5 通过 miR-27a-3p 调节 BNIP3 和 p38 MAPK 通路，通过 miR-27a-3p/BNIP3 通路促进 PaCa 细胞凋亡。体内实验的结果也表明 DGCR5 对 PaCa 裸鼠模型具有积极作用。总的来说，本研究的结果表明，lncRNA DGCR5 通过 miR-27a-3p 上调 BNIP3 和 p38 MAPK 通路，促进 PaCa 细胞凋亡，从而抑制 PaCa 的发展。
