目的:我们使用蛋白质组测序和实验验证来鉴定成釉细胞瘤中潜在的铁凋亡相关蛋白。
方法:收集成釉细胞瘤(n=14)和正常牙龈组织(n=5)的样本进行蛋白质组测序,以鉴定成釉细胞瘤中的差异表达蛋白(DEP)。从FerrDbV2下载与铁凋亡相关的基因,然后将其与DEP进行比较以获得与铁凋亡相关的DEP(FR-DEP)。进行了功能富集分析,建立了蛋白质-蛋白质相互作用网络。使用Cytoscape软件筛选了hub蛋白,并从DrugBank数据库中检索到针对它们的潜在药物。选择了hub蛋白进行免疫组织化学验证,并在成釉细胞瘤中评估其表达,牙源性角化囊肿,牙质囊肿,和正常的牙龈组织.培养原代成釉细胞瘤细胞以探索蛋白质对肿瘤细胞迁移特性的影响。
结果:共筛选了58个FR-DEP,并鉴定了六种hub蛋白:mTOR,NFE2L2,PRKCA,STAT3,EGFR,和CDH1。免疫组织化学分析显示mTOR在成釉细胞瘤中的表达与在牙源性角化囊肿中的表达相比上调。牙质囊肿,和正常的牙龈组织.p-mTOR在成釉细胞瘤中高表达,阳性率为83.3%。此外,雷帕霉素,mTOR的抑制剂,可以抑制原代培养成釉细胞瘤细胞的迁移能力。
结论:我们的结果揭示了成釉细胞瘤中的铁凋亡相关蛋白及其潜在的生物学过程。此外,mTOR过表达,被发现与成釉细胞瘤的侵袭性有关,这可能是未来治疗的潜在目标。
OBJECTIVE: We used proteomic sequencing and experimental verification to identify the potential ferroptosis-related proteins in ameloblastoma.
METHODS: Samples of ameloblastoma (n = 14) and normal gingival tissues (n = 5) were collected for proteomic sequencing to identify differentially expressed proteins (DEPs) in ameloblastoma. Ferroptosis-related genes were downloaded from FerrDb V2, which were then compared with DEPs to obtain ferroptosis-related DEPs (FR-DEPs). A functional enrichment analysis was performed, and a protein-protein interaction network was built. The hub proteins were screened using the Cytoscape software, and potential drugs targeting them were retrieved from the DrugBank database. A hub protein was selected for immunohistochemical validation, and its expression was assessed in ameloblastomas, odontogenic keratocysts, dentigerous cysts, and normal gingival tissues. The primary ameloblastoma cells were cultured to explore the effect of the protein on the migratory properties of the tumour cells.
RESULTS: A total of 58 FR-DEPs were screened, and six hub proteins were identified: mTOR, NFE2L2, PRKCA, STAT3, EGFR, and CDH1. Immunohistochemical analysis showed that mTOR expression was upregulated in ameloblastomas compared with that in odontogenic keratocysts, dentigerous cysts, and normal gingival tissues. p-mTOR was highly expressed in ameloblastomas, with a positivity rate of 83.3%. In addition, rapamycin, an inhibitor of mTOR, can inhibit the migratory capacity of primary cultured ameloblastoma cells.
CONCLUSIONS: Our results revealed the ferroptosis-related proteins in ameloblastomas and their underlying biological processes. Additionally, mTOR was overexpressed and was found to be associated with the aggressiveness of ameloblastomas, which may be a potential target for future treatments.