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目的探讨小剂量维甲酸诱导对神经母细胞瘤(NB)化疗耐药的影响。方法选择人NB细胞株SH SY5Y,用RT PCR方法检测小剂量全反式维甲酸(ATRA)作用下TrkB mRNA水平;通过四甲基偶氮蓝比色(MTT)法及流式细胞仪(FCM)检测ATRA诱导前后顺铂对人NB细胞株SH SY5Y的生长及凋亡的影响。结果SH SY5Y中未检测到TrkB mRNA表达,用1、10、100nM/LATRA诱导5d后可检测到TrkB mRNA表达,且随ATRA浓度增加TrkB mRNA水平逐渐升高。100nM/LATRA诱导7d,TrkB mRNA的相对比值与诱导5d比差异无显著性意义(P>0.05);MTT分析显示:BDNF+顺铂组(无TrkB表达)、ATRA+顺铂组(无BDNF刺激)细胞存活率同单用顺铂组比较无显著性差异(P>0.05);10nM/LATRA+BDNF+顺铂组NB细胞存活率明显高于单用顺铂组(P<0.01);FCM分析显示:ATRA+BDNF+顺铂组NB细胞凋亡率明显低于单用顺铂组(P<0.01)。结论存在BDNF的条件下,小剂量ATRA能阻断顺铂对SH SY5Y的细胞毒性作用,从而使SY5Y细胞对化疗耐药。
Objective To investigate the effect of low-dose retinoid acid on chemoresistance of neuroblastoma (NB). Methods SH SY5Y human NB cell line was selected and TrkB mRNA level was detected by RT-PCR. The expression of TrkB mRNA was detected by MTT assay and flow cytometry (FCM) ) To detect the effect of cisplatin on the growth and apoptosis of human NB cell line SH SY5Y before and after ATRA induction. Results No TrkB mRNA expression was detected in SH SY5Y cells. The expression of TrkB mRNA was detected after treated with 1, 10, 100 nM / L ATA for 5 days, and the level of TrkB mRNA increased gradually with the increase of ATRA concentration. There was no significant difference in the relative ratio of TrkB mRNA to the 5d after induced by 100nM / LATRA (P> 0.05). MTT assay showed that the expression of TrkB in BDNF + cisplatin group (without TrkB expression), ATRA + cisplatin group The survival rate of NB cells in 10nM / LATRA + BDNF + cisplatin group was significantly higher than that in cisplatin alone group (P <0.01). FCM analysis showed that ATRA The apoptosis rate of NB cells in + BDNF + cisplatin group was significantly lower than that in cisplatin alone group (P <0.01). Conclusion Under the condition of BDNF, the low dose of ATRA can block the cytotoxic effect of cisplatin on SH SY5Y, so that SY5Y cells are resistant to chemotherapy.