叶酸受体介导的阿糖胞苷抑制髓母细胞瘤增殖并促其凋亡的体外实验研究  

Folate receptor-targeted cytarabine for inhibiting the proliferation of medulloblastoma and promoting its apoptosis： an in vitro experimental study

作　　者：刘海龙[1] 曲彦明[1] 黄铂渊 宁威海 李翰林[1] 韩松[2] 张宏伟[1] 李俊发[2] 于春江[1] 

机构地区：[1]首都医科大学三博脑科医院神经外科,北京100093 [2]首都医科大学神经生物学系,北京脑重大疾病研究院

出　　处：《中华神经外科杂志》2015年第5期514-518,共5页Chinese Journal of Neurosurgery

基　　金：国家临床重点专科建设资助（SG-2011-03-1-1）

摘　　要：目的 探讨叶酸受体介导的阿糖胞苷（folate receptor targeted cytarabine,FR-Ara-C）对髓母细胞瘤Daoy细胞株的增殖抑制及凋亡促进作用.方法 利用细胞免疫荧光技术及激光共聚焦扫描技术检测叶酸受体α（folate receptors-α,FRs-α）在Daoy细胞株的表达;利用噻唑蓝（MTT）比色法检测不同浓度（0.5 ～ 100.0）＆#215;10-3 mmol/L FR-Ara-C及普通阿糖胞苷（cytarabine,Ara-C）作用于Daoy细胞不同时间（2 ～24 h）后的增殖抑制率并比较其差异;挑选差异最为明显的组别行AnnexinV-FITC/PI细胞凋亡检测.结果 FRs-α在Daoy细胞膜及细胞核内均有表达;相同浓度的FR-Ara-C及Ara-C作用Daoy细胞不同时间时,FR-Ara-C较Ara-C增殖抑制率更高,差异具有统计学意义（P＜0.05）;不同浓度的FR-Ara-C及Ara-C作用Daoy细胞相同时间时,FR-Ara-C较Ara-C增殖抑制率更高,差异具有统计学意义（P＜0.05）;FR-Ara-C作用8h时产生的增殖抑制率较Ara-C效应增高最为明显,此时间点半数抑制浓度（50％ concentration of inhibition,IC50）为最佳作用浓度;流式细胞学分析浓度为50.0＆#215;10-3 mmol/L的FR-Ara-C及Ara-C作用8h时细胞凋亡率为（92.8±;2.3）％,（62.3±;1.5）％;10.0＆#215;10-3 mmol/L的FR-Ara-C及Ara-C作用8h时细胞凋亡率为（87.1±;2.4）％,（44.7±;1.7）％;相同浓度时FR-Ara-C作用后凋亡率较Ara-C明显较高,差异具有统计学意义（P＜0.05）.结论 FR-Ara-C对髓母细胞瘤在体外具有更加明显的增殖抑制及凋亡促进作用.Objective To investigate the folate receptor-targeted cytarabine （FR-Ara-C） for the proliferation inhibition and apoptosis promotion effect of the Daoy medulloblastoma cell lines.Methods The immunofluorescence and confocal laser scanning techniques were used to detect the expression of folate receptors-α （FRs-ot） in Daoy cell lines.Methyl thiazolyl tetrazolium （MTT） colorimetric method was used to detect the proliferation inhibition rate after different concentrations （0.5-100.0 ＆#215; 10-3 mmol/L FR-Ara-C and ordinary cytarabine [Ara-C]） acting on Daoy cells at different times （2-24 h） and their differences were compared.The group with the most obvious difference was selected for conducting AnnexinV-FITC/PI apoptosis detection.Results FRs-α expressed in both Daoy cell membrane and nucleus.When the same concentration of FR-Ara-C and Ara-C acted on Daoy cells at different times,the proliferation inhibition rate of FR-Ara-C was higher than that of Ara-C.There was significant difference （P 〈 0.05）;when the different concentrations of FR-Ara-C and Ara-C acted on Daoy cells at the same time,the proliferation inhibition rate of FR-Ara-C was higher than that of Ara-C.There was significant difference （P 〈 0.05）;when FR-Ara-C acted on for 8 h,the increased inhibition rate was most obvious compared with the Ara-C effect,50% concentration of inhibition （ICS0） at this time point was the optimal action concentration.Flow cytometry analysis showed that when 50 ＆#215; 10-3 mmol/L concentration of FR-Ara-C and Ara-C acted on for 8 h,the apoptosis rates were 92.8 ±; 2.3% and 62.3 ±; 1.5%;when 10.0 ＆#215; 10-3 mmol/L concentration of FR-Ara-C andAra-C acted on for 8 h,the apoptosis rates were 87.1 ±;2.4% and 44.7 ±; 1.7%;when the concentration was the same,the apoptosis rate was significantly higher than that of Ara-C after the action of FR-Ara-C.There was significant difference （P 〈 0.05）.Condution FR-Ara-C has more significant proliferation inhibition and apoptosis promotion

关 键 词：髓母细胞瘤 叶酸盐受体1 阿糖胞苷 半数抑制浓度 

分 类 号：R739.4[医药卫生—肿瘤]