CAJ | 학술논문

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)×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50.0×10-3 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)％;10.0×10-3 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.
목적 탐토협산수체개도적아당포감(folate receptor targeted cytarabine,FR-Ara-C)대수모세포류Daoy세포주적증식억제급조망촉진작용.방법 이용세포면역형광기술급격광공취초소묘기술검측협산수체α(folate receptors-α,FRs-α)재Daoy세포주적표체;이용새서람(MTT)비색법검측불동농도(0.5 ～ 100.0)×10-3 mmol/L FR-Ara-C급보통아당포감(cytarabine,Ara-C)작용우Daoy세포불동시간(2 ～24 h)후적증식억제솔병비교기차이;도선차이최위명현적조별행AnnexinV-FITC/PI세포조망검측.결과 FRs-α재Daoy세포막급세포핵내균유표체;상동농도적FR-Ara-C급Ara-C작용Daoy세포불동시간시,FR-Ara-C교Ara-C증식억제솔경고,차이구유통계학의의(P＜0.05);불동농도적FR-Ara-C급Ara-C작용Daoy세포상동시간시,FR-Ara-C교Ara-C증식억제솔경고,차이구유통계학의의(P＜0.05);FR-Ara-C작용8h시산생적증식억제솔교Ara-C효응증고최위명현,차시간점반수억제농도(50％ concentration of inhibition,IC50)위최가작용농도;류식세포학분석농도위50.0×10-3 mmol/L적FR-Ara-C급Ara-C작용8h시세포조망솔위(92.8±2.3)％,(62.3±1.5)％;10.0×10-3 mmol/L적FR-Ara-C급Ara-C작용8h시세포조망솔위(87.1±2.4)％,(44.7±1.7)％;상동농도시FR-Ara-C작용후조망솔교Ara-C명현교고,차이구유통계학의의(P＜0.05).결론 FR-Ara-C대수모세포류재체외구유경가명현적증식억제급조망촉진작용.
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 × 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 × 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 × 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 effect on medulloblastoma in vitro.