MSAA3蛋白功能片段及其修饰肽对HT29细胞MUC3表达的影响

Influence of MSAA3 protein fragment and modified peptide thereof on the MUC3 expression of HT29 cells

目的观察MSAA3蛋白功能片段TFLK合成肽(10-TFLK)及其MAP4修饰肽对高分化状态HT29细胞表达黏蛋白3(MUC3)的影响。方法将10-TFLK和10-TFLK修饰肽以不同浓度(0、50、100、200、400、800μg/ml)和时间(0.5、1、3h)分别刺激促分化后的HT29细胞,采用实时荧光定量PCR检测各组细胞MUC3mRNA的表达变化,并用Western blotting分析不同肽刺激后MUC3蛋白的表达量。结果 HT29细胞MUC3mRNA表达量随处理浓度和时间的不同而不同,以200μg/ml刺激1h后的表达量为最高。10-TFLK刺激后,MUC3mRNA表达量是对照组的3.22±0.24倍,修饰肽10-TFLK-MAP刺激后,MUC3mRNA表达量是对照组的7.37±0.81倍(P<0.01)。10-TFLK和修饰肽10-TFLK-MAP处理后MUC3蛋白表达量分别是对照组的1.52±0.22和4.72±0.51倍(P<0.01)。结论 200μg/ml修饰肽10-TFLK-MAP作用1h对HT29细胞MUC3表达的刺激效果显著强于10-TFLK。MAP4修饰肽可优先应用于动物模型试验中。

Objective To investigate the effect of MSAA3 protein fragment（10-TFLK） and its MAP4 modified peptide（10-TFLK-MAP） on the expression of MUC3 in enterocyte-like HT29 cells.Methods The enterocyte-like HT29 cells were treated with 10-TFLK and 10-TFLK-MAP at the doses of 0,50,100,200,400 and 800 μg/ml for 0.5,1 and 3 hours.The expressions of MUC3 mRNA and protein were detected by real time PCR and Western blotting.Results HT29 cells went to the highest expression of MUC3 mRNA when treated with 10-TFLK at the dose of 200μg/ml for one hour.When stimulated by 10-TFLK or 10-TFLK-MAP,the expression of MUC3 mRNA increased by 3.22±0.24 and 7.37±0.81 folds respectively,and of MUC3 protein increased by 1.52±0.22 and 4.72±0.51 folds respectively,compared with the control group（P0.01）.Conclusions The modified peptide 10-TFLK-MAP can up-regulate MUC3 expression of HT29 cells,the effect of which is more increased than by 10-TFLK at the dose of 200μg/ml for one hour.MAP4 modified peptide should be used in animal model trial as a first choice.