胰岛素联合5-氟尿嘧啶对Eca-109细胞的化疗增效作用

Enhancement of chemotherapeutic response of insulin and 5-Fluorouracil on Eca-109 cell

目的:观察胰岛素对化疗药物5-氟尿嘧啶(5-Fu)的最佳增效作用时间和浓度,并探讨其发挥化疗增效作用的相应受体。方法:采用MTT方法检测胰岛素、5-Fu和胰岛素受体(IR)抗体、胰岛素样生长因子Ⅰ受体(IGF-ⅠR)抗体对食管癌Eca-109细胞的生长抑制作用。胰岛素作用时间分别设5-Fu作用前0、4、8、12、24h和作用后4、8h,胰岛素作用浓度分别设0.08、0.80、1.60、8.00、16.00、80.00、160.00、200.00U/L,IR及IGF-ⅠR抗体单独作用时分别设1∶50、1∶100、1∶200、1∶400、1∶800、1∶1600稀释度,2种抗体联合作用时分别设1∶50、1∶100、1∶200稀释度。结果:胰岛素于5-Fu作用前0～8h加入可发挥化疗增效作用(P均<0.01),其中以提前4h加入增效作用最佳(P<0.05)。胰岛素的含量在0.80～8.00U/L时增效作用随着胰岛素剂量的增加而增加(P<0.05),但当胰岛素剂量在16.00～200.00U/L之间时,细胞抑制率并未随着胰岛素剂量的增加而增加,而是稳定在一定的水平。除1∶800和1∶1600稀释度外,抗体阻断IR后,胰岛素联合化疗组与单一化疗组间细胞生长抑制率差异无统计学意义(P均>0.05)。而采用抗体阻断IGFⅠ-R后,胰岛素联合化疗组细胞生长抑制率高于单用化疗组(P均<0.05)。结论:胰岛素以8U/L于5-Fu作用前4h加入化疗增效作用最佳,胰岛素主要通过IR发挥增效作用。

Aim： To observe the best enhancement time and concentration of insulin on chemotherapeutic response of 5-Fluorouraeil （5-Fu） and explore the corresponding receptor by which insulin exerts the enhancement. Methods： MTT method was used to test the growth inhibitory rate of esophageal cancer cell Eea-109 acted by insulin and 5-Fu and different antibody. The time point of insulin was 0,4,8,12,24 h befere 5-Fu, and 4,8 h after 5-Fu. The concentration of insulin was 0.08,0.80,1.60,8.00,16.00,80.00,160.00,200.00 U/L. The concentration of IR and IGF- I R antibody was 1 ： 50, 1 ： 100,1 ： 200,1 ： 400,1 ： 800,1 ： 1 600. when IR and IGF-[ R antibody added together, their concentration was 1 ： 50,1 ： 100,1 ： 200. Results： Insulin could exert the enhancement on chemotherapeutic response of 5-Fu when insulin was applied 0 - 8 h before 5-Fu（ P 〈0.01 ）. The enhancement was the best when insulin was added 4 h before 5-Fu（ P 〈 0.05）. The enhancement increased with insulin dosage augment when the concentration of insulin varied between 0.80 ~ 8.00 U/L, which was the best with 8 U/L（ P 〈 0.05 ）. There was no significance on the inhibitory rate of cell growth be- tween 5-Fu alone group and IR antibody combination groups except for the dilution concentration 1 ： 800 and 1 ： 1 600 （ P 〉 0.05 ）. The inhibitory rate of cell growth in IGF- I R antiboby combination groups was significantly higher than that in 5-Fu alone group（P 〈0.05）. Gonclusion： Insulin could exert the best enhancement with 8 U/L when added 4 h before 5- Fu. Insulin receptor plays an important role in the enhancement of insulin on chemotherapeutic response of 5-Fu.