探讨Circ_0000253在骨肉瘤中的诊断价值

The role of Circular RNA circ_0000253 in diagnosis and prognosis of osteosarcoma

目的探讨circ_0000253作为骨肉瘤(osteosarcoma,OS)诊断和预后生物标志物的价值。方法GEO数据库筛选OS中差异表达的circRNAs,预测和circRNA存在结合位点的微小RNA(MicroRNA,miRNA)进一步构建ceRNA网络,GO和KEGG分析筛选靶基因。qRT-PCR检测112例OS患者癌组织和癌旁组织、OS细胞及正常成骨细胞中circ_0000253、miR-578及TGFβ2的表达水平。分析circ_0000253对OS的诊断价值及其对预后的影响,探讨circ_0000253表达水平与患者临床病理参数的关系。进一步干预细胞中circ_0000253和miR-578的表达,流式细胞术检测细胞凋亡,MTT实验检测细胞增殖活力。结果OS组织中circ_0000253表达水平显著高于癌旁组织(t=11.17,P<0.001)。circ_0000253表达与Enneking分期和组织分级和远处转移显著相关(均P<0.05)。circ_0000253可作为OS的一个有效诊断指标(AUC=0.84,P<0.001)且高表达circ_0000253的患者预后较差。Circ_0000253可能通过miR-578/TGFβ2网络对OS进行调控。敲减circ_0000253能降低细胞增殖活力,诱导细胞凋亡,而该作用能被miR-578 inhibitor部分挽救(均P<0.05)。结论circ_0000253可作为OS的新的诊断和预后生物标志物,对OS的靶向治疗也有积极意义。

Objective To evaluate the value of circ_0000253 as a biomarker for(osteosarcoma,OS)diagnosis and prognosis.Methods GEO database was used to screen the differentially expressed circRNAs in OS,microRNAs(miRNAs)with binding sites of circRNA were predicted to further construct the ceRNA network,and GO and KEGG analysis were used to screen the target genes.qRT-PCR was used to detect the expression of circ_0000253,miR-578 and TGFβ2 in cancer and paracancerous tissue in OS patients,as well as OS cells and normal osteoblasts.The diagnostic value of circ_0000253 for OS and its impact on prognosis were analyzed,and the relationship between circ_0000253 expression level and clinicopathological parameters of patients was discussed.Further the expression of circ_0000253 and miR-578 in cells was intervened,apoptosis was detected by flow cytometry and cell proliferation was detected by MTT assay.Results The expression of circ_0000253 in OS tissue was higher than that in paracancerous tissue(t=11.17,P<0.001).The expression of circ_0000253 was significantly correlated with Enneking stage,histopathology grade and distant metastasis(all P<0.05).Circ_0000253 can be used as an effective diagnostic index of OS(AUC=0.84,P<0.001)and the prognosis of patients with high expression of circ_0000253 is poor.Circ_0000253 may regulate OS through miR-578/TGFβ2 network.Knockdown of circ_0000253 reduced cell proliferation and induced apoptosis,which could be partially saved by miR-578 inhibitor(all P<005).Conclusion circ_0000253 may serve as a new biomarker for OS diagnosis and prognosis,which has a positive meaning for targeted therapy of OS.