Yuan Bo, Xu Yimin, Zheng Shaoqin
Department of Neurosurgery, The First Affiliated Hospital, Shantou University Medical College, No. 57, Changping Road, Shantou, 515041, Guangdong, People's Republic of China.
Mol Cell Biochem. 2022 Feb;477(2):549-557. doi: 10.1007/s11010-021-04289-w. Epub 2021 Nov 29.
Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 (PLOD1) is a collagen-related lysyl hydroxylase and its prognostic value in glioma patients was verified. However, its biological function in glioma has yet to be fully investigated. The PLOD1 mRNA status and clinical significance in gliomas were assessed via the GEPIA database. Overexpression or targeted depletion of PLOD1 was carried out in the human glioma cell line U87 and verified by western blotting. CCK8 and colony formation assays were implemented to examine the impact of PLOD1 on the proliferative and colony-forming phenotypes of U87 cells. Luciferase reporter assays and HSF1-specific pharmacologic inhibitors (KRIBB11) were employed to determine the regulatory relationship between PLOD1 and heat shock factor 1 (HSF1). High expression of PLOD1 was observed in tissue samples of glioblastoma multiforme (GBM) and brain lower-grade glioma (LGG). GEPIA overall survival further demonstrated that both GBM and LGG patients with high PLOD1 displayed worse clinical outcomes compared with those with low PLOD1. Overexpression and targeted depletion of PLOD1 enhanced and suppressed U87 cell proliferation and colony formation, respectively. Luciferase reporter assays showed that PLOD1 significantly enhanced the transcriptional activity of HSF1 in HEK293T cells. PLOD1 deficiency in U87 cells inhibited HSF1-induced survivin accumulation, whereas KRIBB11 also blocked the PLOD1-overexpressing induced survivin expression. An inhibitor of HSF1 signaling events abolished the increased clonogenic potential caused by PLOD1 overexpression in U87 cells. High expression of PLOD1 can increase the proliferation and colony formation of U87 cells by activating the HSF1 signaling pathway. This study suggested PLOD1/HSF1 as an effective therapeutic target for gliomas.
原胶原赖氨酸2-氧代戊二酸5-双加氧酶1（PLOD1）是一种与胶原相关的赖氨酰羟化酶，其在胶质瘤患者中的预后价值已得到验证。然而，其在胶质瘤中的生物学功能尚未得到充分研究。通过GEPIA数据库评估了PLOD1在胶质瘤中的mRNA状态及临床意义。在人胶质瘤细胞系U87中进行PLOD1的过表达或靶向敲减，并通过蛋白质免疫印迹法进行验证。采用CCK8和集落形成实验检测PLOD1对U87细胞增殖和集落形成表型的影响。利用荧光素酶报告基因实验和HSF1特异性药理抑制剂（KRIBB11）确定PLOD1与热休克因子1（HSF1）之间的调控关系。在多形性胶质母细胞瘤（GBM）和低级别脑胶质瘤（LGG）的组织样本中观察到PLOD1的高表达。GEPIA总生存分析进一步表明，与PLOD1低表达的患者相比，PLOD1高表达的GBM和LGG患者的临床结局更差。PLOD1的过表达和靶向敲减分别增强和抑制了U87细胞的增殖和集落形成。荧光素酶报告基因实验表明，PLOD1显著增强了HEK293T细胞中HSF1的转录活性。U87细胞中PLOD1的缺失抑制了HSF1诱导的生存素积累，而KRIBB11也阻断了PLOD1过表达诱导的生存素表达。HSF1信号事件抑制剂消除了U87细胞中PLOD1过表达导致的克隆形成潜力增加。PLOD1的高表达可通过激活HSF1信号通路增加U87细胞的增殖和集落形成。本研究提示PLOD1/HSF1是胶质瘤的有效治疗靶点。
