Foxp3在神经母细胞瘤LA-N-6细胞的表达及化疗药物对其影响

Neuroblastoma LA-N-6 cells express Foxp3 which can be suppressed by chemotherapeutic agents

目的研究神经母细胞瘤细胞是否表达免疫逃逸相关分子Foxp3及其对化疗药物的敏感性。方法体外培养神经母细胞瘤细胞株LA-N-6;MTT试验明确化疗药物环磷酰胺(CTX)、盐酸吡柔比星(THP)对LA-N-6的效应剂量,流式细胞术(FACS)及实时定量PCR(RT-PCR)测定Foxp3在LA-N-6中的表达和CTX、THP对LA-N-6中Foxp3表达的影响。结果 FACS结果显示LA-N-6细胞高表达Foxp3,加入CTX、THP后显著降低了LA-N-6上Foxp3的表达(P<0.05);RT-PCR显示CTX显著降低了LA-N-6细胞中Foxp3的表达(P<0.05),THP加药组与对照组比较差异无统计学意义。结论肿瘤逃逸相关转录因子Foxp3高表达于神经母细胞瘤细胞LA-N-6;CTX、THP能分别在蛋白或基因水平降低Foxp3分子在LA-N-6的表达,提示CTX、THP可能通过抑制Foxp3分子的表达而抗肿瘤的作用。

Objective To investigate whether neuroblastoma cells LA-N-6 express Foxp3 and whether the expression of Foxp3 is sensitive to chemotherapy by cyclophosvnamide（CTX）and pirarubicin（THP）.Methods Expression of Foxp3 on LA-N-6 cells was examined by flow cytometry analysis.The dose-effects of chemotherapy drugs including CTX and THP on LA-N-6 cells were investigated by MTT assay.The effects of CTX and THP on Foxp3 expression were examined by flow cytometry and real-time PCR assays.Results Flow cytometry analysis showed that LA-N-6 cells expressed Foxp3 at a high level.At sub-optimal concentration,chemotherapy drugs CTX and THP significantly down-regulated expression of Foxp3 on LA-N-6 cells at protein level（P0.05）.CTX also decreased the expression of Foxp3 at mRNA level（P0.05）.Conclusions Neuroblastoma cells LA-N-6 express Foxp3 at a high level,which can be suppressed by chemotherapy drugs CTX and THP.These data suggest that chemotherapy might suppress the growth and metastasis of tumor cells partially through inhibiting the expression of Foxp3.