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目的观察环磷酰胺对儿童纵隔神经母细胞瘤LAN-5和SK-N-SH细胞CXC趋化因子受体4(CXCR4)和叉头蛋白3(Foxp3)表达的影响。方法采用实时定量聚合酶链反应(Real-time PCR)法检测CXCR4和Foxp3基因mRNA的相对表达,MTT法检测细胞增殖,流式细胞仪分析CXCR4和Foxp3基因的表达。结果CXCR4和Foxp3在LAN-5和SK-N-SH细胞高表达;环磷酰胺对LAN-5和SK-N-SH细胞的IC50分别为6.5μmol/L和3.9μmol/L;环磷酰胺显著降低了LAN-5细胞表面CXCR4基因蛋白的表达(P<0.01),也显著降低了SK-N-SH细胞CXCR4 mRNA的表达(P<0.05),同时环磷酰胺显著下调Foxp3基因蛋白(P<0.05)和Foxp3 mRNA(P<0.01)在LAN-5细胞的表达。结论 CXCR4和Foxp3可能为神经母细胞瘤化疗的潜在靶点。
Objective To observe the effect of cyclophosphamide on the expression of CXCR4 and Foxp3 in the mediastinal neuroblastoma LAN-5 and SK-N-SH in children. Methods Real-time PCR was used to detect the mRNA expression of CXCR4 and Foxp3, the proliferation of cells was detected by MTT assay and the expression of CXCR4 and Foxp3 was analyzed by flow cytometry. Results CXCR4 and Foxp3 were highly expressed in LAN-5 and SK-N-SH cells. The IC50 of cyclophosphamide in LAN-5 and SK-N-SH cells were 6.5μmol / L and 3.9μmol / L, (P <0.01), but also decreased the expression of CXCR4 mRNA in SK-N-SH cells (P <0.05), while cyclophosphamide significantly down-regulated Foxp3 protein (P < 0.05) and Foxp3 mRNA (P <0.01) in LAN-5 cells. Conclusion CXCR4 and Foxp3 may be potential targets for the chemotherapy of neuroblastoma.