【摘要】 目的 探讨右美托咪定（Dex）对骨肉瘤（OS）细胞的影响及其作用机制。方法 通过MTT实验确定Dex、α2-肾上腺素受体（α2-AR）拮抗剂、PKA抑制剂或ERK1/2抑制剂的最佳暴露浓度；从GEPIA数据库提取与OS样本中ERK1/2表达呈正相关且可能介导其促瘤作用的基因，并采用qRT-PCR、Western blot验证其在OS细胞中的表达情况；利用siRNA沉默候选基因后，通过MTT、流式细胞术、Transwell及集落形成实验，评估ERK1/2通路基因在Dex影响OS细胞增殖、凋亡、侵袭与集落形成能力过程中的作用。结果 25 nM Dex是促进OS细胞活力的最佳浓度，ERK1/2抑制剂（Ravoxertinib）可显著拮抗低浓度Dex导致的OS细胞活力增强，而抑制α2-AR或PKA效果较弱。生信分析发现TRIO、PKN2、CKAP5和NEK4与OS中ERK1/2致癌作用高度相关，Western blot显示Ravoxertinib可阻断Dex引起的TRIO、PKN2、CKAP5和NEK4表达上调。沉默TRIO、PKN2或CKAP5能有效抑制Dex诱导的OS细胞活力增强、侵袭能力提升及集落形成增加，同时减弱Dex对细胞凋亡的抑制作用。结论 低浓度Dex通过ERK1/2依赖性方式上调TRIO、PKN2和CKAP5的表达，从而促进OS细胞的恶性生物学潜能。更多还原

【Abstract】 Objective To investigate the effect of Dexmedetomidine(Dex) on osteosarcoma(OS)cells and its underlying mechanism.Methods The optimal exposure concentrations of Dex,a2-adrenergic receptor(a2-AR) antagonist,PKA inhibitor,and ERK1/2 inhibitor were determined by MTT assay.Genes positively correlated with ERK1/2 expression in OS samples and potentially mediating its tumor-promoting effect were extracted from the GEPIA database,and their expression levels in OS cells were verified by qRTPCR and Western blot.After silencing candidate genes with siRNA,MTT assay,flow cytometry,Transwell assay,and colony formation assay were performed to evaluate the role of ERK1/2 pathway genes in the processes by which Dex affects OS cell proliferation,apoptosis,invasion,and colony formation ability.Results 25 nM Dex was the optimal concentration for promoting OS cell viability.The ERK1/2 inhibitor(Ravoxertinib) significantly antagonized the enhancement of OS cell viability induced by low-concentration Dex,whereas inhibition of a2-AR or PKA exerted weaker effects.Bioinformatics analysis identified that TRIO,PKN2,CKAP5,and NEK4 were highly associated with the oncogenic function of ERK1/2 in OS.Western blot demonstrated that Ravoxertinib blocked the upregulation of TRIO,PKN2,CKAP5,and NEK4 induced by Dex.Silencing TRIO,PKN2,or CKAP5effectively suppressed Dex-induced increases in OS cell viability,invasion capacity,and colony formation,while attenuating the inhibitory effect of Dex on cell apoptosis.Conclusion Low-concentration Dex upregulates the expression of TRIO,PKN2,and CKAP5 in an ERK1/2-dependent manner,thereby promoting the malignant biological potential of OS cells.更多还原