顺铂及依托泊苷对神经母细胞瘤细胞中Trk基因家族表达的影响及临床意义

摘要

目的 神经母细胞瘤(NB)是儿童常见的实体瘤,而三种不同Trk基因的表达变化与NB的预后及转归关系密切;为研究如何优化化疗药物疗效以提高NB患儿的生存率,本文检测了顺铂(DDP)及依托泊苷(VP16)对人NB的SK-N-SH细胞系的杀伤作用及对细胞中Trk基因家族表达的影响.方法 通过CCK-8药物诱导细胞毒性实验检测不同浓度梯度的DDP及VP16对SK-N-SH细胞的杀伤作用以及半数杀伤浓度(IC50);用qRT-PCR方法检测DDP及VP16在不同浓度下分别作用24 h及48 h后,对SK-N-SH细胞中TrkA、TrkB和TrkC基因表达变化.结果 (1)DDP及VP16药物浓度的增加与其对SK-N-SH细胞的杀伤作用呈线性关系;且48 h比24 h杀伤作用更明显.(2)SK-N-SH细胞中TrkA及TrkC的表达均随着DDP浓度和作用时间的增加而上调,尤其在10 μg/mL浓度、24h最明显;而TrkB的表达在24 h时随着DDP浓度增加而上调,48 h时该促进作用不明显;(3)SK-N-SH细胞中TrkA、TrkB及TrkC的表达亦随着VP16浓度和作用时间的增加而上调.结论 DDP及VP16对NB细胞SK-N-SH有明显杀伤作用并对其Trk基因家族的表达有影响.%Objective Neuroblastoma (NB) is a common childhood tumor.The various expressions of the Trk family in different stages are clearly related with the prognosis and outcome of NB.To study how to optimize the chemotherapeutics curative effect improving survival rates,we have tested the drug toxicity and half maximal inhibitory concentration (IC50) for Cisplatin (DDP) and etoposide (VP16) and the influence on gene expression of TrkA,TrkB and TrkC in NB cell line SK-N-SH cell.Methods SK-N-SH cells were cultured to examine the drug toxicity and IC50 through CCK-8 druginduced cytotoxicity assays,then TrkA,TrkB and TrkC gene expression were detected by Real-time RT PCR,cells were cultured for 24 h and 48 h respectively.Results ① With the increase in concentration of DDP and VP16,the number of survival SK-N-SH cell is significantly reduced.There is linear relation between concentration and killing effect.The cells treated for 24h were more significantly reduced with treatment for 48h.② With increasing concentration and time of DDP,the expression of TrkA and TrkC were all significantly upregulated,especially in 10μg/mL at 24h were significantly higher than the other.With increasing concentration of DDP,the expression of TrkB was significantly upregulated at 24h,not at 48h.③ With increasing concentration and time of VP16,the expression of TrkA、TrkB and TrkC were all significantly upregulated.Conclusions DDP and VP16 have significant killing effect on SK-N-SH cells,with different impactonTrk family.