Jin Fei, Long Kaibing, Zhang Jian
Department of Orthopedics, Hunan Aerospace Hospital, No. 189, Fenglin Third Road, Yuelu District, Changsha, 410205, Hunan, China.
Department of Orthopedics, The First Affiliated Hospital of University of South China, Hengyang, Hunan, China.
Mol Biol Rep. 2025 Jun 28;52(1):647. doi: 10.1007/s11033-025-10741-7.
Osteosarcoma is a malignant tumor in bone and soft tissue tumors, characterized by high invasiveness and low survival rate. Here, we studied the effect of cAMP-responsive element binding protein 1 (CREB1) on osteosarcoma cell proliferation and invasion.
mRNA levels of CREB1, miRNA-183-5p and prospero homeobox 1 (PROX1) were detected by quantitative real-time polymerase chain reaction (RT-qPCR), while CREB1 and PROX1 protein levels were evaluated utilizing Western blot in osteosarcoma cell lines and tissues. Immunohistochemistry investigated CREB1 expression in osteosarcoma tissues. Colony formation, transwell and flow cytometry experiments detected cell proliferation, invasion and apoptosis, respectively. Dual luciferase reporter assay, chromatin immunoprecipitation (ChIP) assay and RNA immunoprecipitation (RIP) assay verified target relationship between CREB1, miRNA-183-5p and PROX1.
CREB1 expression was increased in osteosarcoma tissues and cells. Reduced CREB1 inhibited osteosarcoma cell proliferation and invasion, and promoted apoptosis. Moreover, CREB1 targets miRNA-183-5p and transcriptionally inhibited miRNA-183-5p expression in osteosarcoma cells. Additionally, PROX1 was a target gene of miRNA-183-5p, where expression was inhibited by miRNA-183-5p. PROX1 upregulation reversed inhibitory effect of miRNA-183-5p overexpression on osteosarcoma cell malignant phenotype.
Downregulation of CREB1 inhibited osteosarcoma cell proliferation and invasion but induced apoptosis through miRNA-183-5p/PROX1 axis. CREB1 may become a new target for diagnosis and treatment of osteosarcoma.
骨肉瘤是一种发生于骨和软组织的恶性肿瘤，具有高侵袭性和低生存率的特点。在此，我们研究了环磷腺苷反应元件结合蛋白1（CREB1）对骨肉瘤细胞增殖和侵袭的影响。
采用定量实时聚合酶链反应（RT-qPCR）检测骨肉瘤细胞系和组织中CREB1、miRNA-183-5p和prospero同源盒蛋白1（PROX1）的mRNA水平，同时利用蛋白质免疫印迹法评估CREB1和PROX1蛋白水平。免疫组织化学检测骨肉瘤组织中CREB1的表达。集落形成实验、Transwell实验和流式细胞术实验分别检测细胞增殖、侵袭和凋亡情况。双荧光素酶报告基因检测、染色质免疫沉淀（ChIP）实验和RNA免疫沉淀（RIP）实验验证CREB1、miRNA-183-5p和PROX1之间的靶向关系。
CREB1在骨肉瘤组织和细胞中表达升高。CREB1表达降低抑制了骨肉瘤细胞的增殖和侵袭，并促进了细胞凋亡。此外，CREB1靶向miRNA-183-5p，并在转录水平上抑制骨肉瘤细胞中miRNA-183-5p的表达。另外，PROX1是miRNA-183-5p的靶基因，其表达受到miRNA-183-�p的抑制。PROX1上调可逆转miRNA-183-5p过表达对骨肉瘤细胞恶性表型的抑制作用。
CREB1下调通过miRNA-183-5p/PROX1轴抑制骨肉瘤细胞增殖和侵袭，但诱导细胞凋亡。CREB1可能成为骨肉瘤诊断和治疗的新靶点。
