潘生丁抑制平衡型核苷转运蛋白对5氟尿嘧啶在胰腺癌细胞内浓聚的影响  

Effect of Inhibiting Equilibrative Nucleoside Transport Proteins with Dipyridamole on 5-FU Intracellular Accumulation in Pancreatic Cancer Cell

作　　者：刘胜利[1] 王晶敏[1] 鞠煌先[2] 朱广华[3] 杜丹[2] 朱春富[1] 徐皓[1] 黄海[1] 

机构地区：[1]东南大学医学院附属中大医院,南京210009 [2]南京大学化学化工学院,南京210009 [3]东南大学公共卫生学院,南京210009

出　　处：《中国药科大学学报》2003年第6期544-548,共5页Journal of China Pharmaceutical University

基　　金：铁道部科技基金资助项目 (No J2 0 0 0Z0 94 )

摘　　要：目的 :研究阻断平衡型核苷转运蛋白 (ENTs)后对胰腺癌细胞膜转运 5氟尿嘧啶 (5 FU)能力的影响。方法 :培养的PANC 1细胞分为ENTs阻断组和ENTs非阻断组 ,阻断组用潘生丁将ENTs阻断 ,非阻断组则未用潘生丁。两组培养基中均加入 5 FU 10 0 μg/ml。温育 15,3 0min ,1,2 ,4h后细胞用橡皮刮取收集。取等量细胞刮取液 2份各 10 0 μl ,一份用毛细管区带电泳测定细胞内 5 FU总量、另一份用作平行样本测定总蛋白含量。根据平行样本细胞总蛋白来确定细胞数 ,再依细胞体积计算单细胞内药物浓度。结果 :ENTs非阻断组细胞内 5 FU浓度在 15min内上升较快 ,达 66 99± 10 16pg/nl,为培养基中 5 FU浓度的 67% ,3 0min为 83 75± 5 3 3 3pg/nl,1h达高峰 (92 86± 1 60 4pg/nl) ,为培养基中 5 FU浓度的 93 % ,以后处于平台。ENTs阻断组细胞内 5 FU浓度上升较慢 ,15min仅为ENTs非阻断组浓度的 68 65% ;3 0min为ENTs非阻断组浓度的 70 66% ;1h达ENTs非阻断组的最高浓度 ;但此后ENTs阻断组细胞内 5 FU浓度仍继续升高 ,2h达 13 8 3± 9 769pg/nl,为ENTs非阻断组的1 64倍 (P <0 0 1)。结果提示 ,完全阻断ENTs后早期细胞内 5 FU浓度上升较慢 ,但随着细胞在 5 FU内暴露时间延长 ,5 FU在细胞内浓聚作用明显。结论 :阻断胰腺?AIM:To investigate the effect of block i ng ENTs on the membrane of pancreatic cancer cell on 5-FU intracellular accumul aion METHOD: Cultured PANC-1 cells were divided into two groups,the ENTs inhibited group and the ENTs non-inhibited group I n ENTs inhibited group the ENTs on cells were inhibited by dipyridamole,while in ENTs non-inhibited group they were not inhibited by any drug Cells in t he two groups were cultured in the medium with 5-Fu (100 μg/ml) Cells were harvested w ith rubber policeman after 15,30 min,1,2,4 h incubation,respectively Afte rwards,two 100 μl cell suspensions were spliced, one was for 5-FU detection with capil l ary zone electrophoresis and the another parallel one for protein assay Cell numb er in its parallel suspensions was defined according to the standard curve of to tal protein vs cell number Then content of 5-FU in a single cell was det ermined a nd intracellular 5-FU concentration in a single cell was finally counted RESULT:In ENTs non-inhibited group,5-FU concentration in cells was i ncr eased quickly to 66 99±10 16 μg/ml within the first 15 min and 83 75±5 3 3 μ g/ml in 30 min The maximal intracellular 5-FU concentration was 92 86±1 60 4 μ g/ml at 60 min Afterwards,the concentration curve produced a platform In E NTs inhibited group,however,the intracellular 5-FU concentration was increased sl owly in early stage It was only 68 65% and 70 66% of that in ENTs non-inhabited group at 15 min and 30 min, respectively. The concention of 5-FU equals to the maximal concentration of that in ENTs non-inhabited group Afterwards, the intracellular 5-FU concentration was continously accumulated in cells with the maximal concentration of 138 3±9 769 μg/ml at 2 h,as 1 64 times as that in ENTs non-inhibited group (P< 0 01) These results implicated that after ENTs tot ally inhibited by dipyridamole the 5-FU intracellular concentration rises slowl y in early stage and furthermore significantly accumulates in cell as the time pro l onged CONCL

关 键 词：胰腺癌 核苷转运蛋白 5-氟尿嘧啶 潘生丁 毛细管区带电泳 

分 类 号：R735.9[医药卫生—肿瘤]