Cancer Chemoradiation Research Group, Department of Surgery, Trinity St. James's Cancer Institute, Trinity Translational Medicine Institute, St. James's Hospital, D08 W9RT 8 Dublin, Ireland.
Cells. 2022 Jul 25;11(15):2294. doi: 10.3390/cells11152294.
Radioresistance remains a significant challenge in treating pancreatic ductal adenocarcinoma (PDAC), contributing to the poor survival rates of this cancer. MicroRNAs (miRs) are small non-coding RNA molecules that may play an essential role in regulating radioresistance by altering the levels of oxidative stress. In this study, we investigated the role and potential mechanisms linking miR-31 to PDAC radioresistance. A pCMV-miR vector containing a miR-31 mimic was stably expressed into a miR-31-deficient PDAC cell line, BxPC-3. Additionally, a pmiRZip lentivector suppressing miR-31 was stably expressed in a miR-31 abundant PDAC cell line, Panc-1. Clonogenic assays were conducted to explore the role of miR-31 manipulation on radiosensitivity. Fluorometric ROS assays were performed to quantify ROS levels. The expression of potential miR-31 targets was measured by Western blot analysis. It was found that the manipulation of miR-31 altered the radiosensitivity in PDAC cells by regulating oxidative stress. Using online bioinformatics tools, we identified the 3'UTR of as a predicted target of miR-31. Our study demonstrates, for the first time, that manipulating miR-31 alters GPx8 expression, regulating ROS detoxification and promoting either a radioresistant or radiosensitive phenotype. MiR-31 may represent a promising therapeutic target for altering radiosensitivity in PDAC cells.
放射抵抗仍然是治疗胰腺导管腺癌 (PDAC) 的重大挑战，导致这种癌症的生存率低。微小 RNA (miRs) 是小的非编码 RNA 分子，通过改变氧化应激水平，可能在调节放射抵抗方面发挥重要作用。在这项研究中，我们研究了 miR-31 与 PDAC 放射抵抗之间的关系及其潜在机制。一个包含 miR-31 模拟物的 pCMV-miR 载体被稳定地表达到 miR-31 缺陷的 PDAC 细胞系 BxPC-3 中。此外，一个抑制 miR-31 的 pmiRZip 慢病毒载体被稳定地表达在 miR-31 丰富的 PDAC 细胞系 Panc-1 中。克隆形成实验被用来探讨 miR-31 操作对放射敏感性的作用。荧光 ROS 分析被用来定量 ROS 水平。潜在的 miR-31 靶标的表达通过 Western blot 分析来测量。结果发现，miR-31 的操作通过调节氧化应激改变了 PDAC 细胞的放射敏感性。使用在线生物信息学工具，我们鉴定出作为 miR-31 预测靶点的 3'UTR。我们的研究首次表明，操纵 miR-31 改变了 GPx8 的表达，调节了 ROS 解毒，促进了放射抵抗或放射敏感表型。miR-31 可能代表改变 PDAC 细胞放射敏感性的有前途的治疗靶点。
