基础医学与临床2013，Vol.33Issue(9)：1199-1204,6.
生长因子受体结合蛋白-2抑制肠癌细胞HT29增殖
Growth factor receptor bound protein-2 inhibits colon cancer HT29 cell proliferation
陈丽妹 1叶韵斌 2刘枋 3陈慧菁 1李洁羽 2苏光建 4林巧燕1
作者信息
- 1. 福建医科大学研究生教育学院,福建福州350014
- 2. 福建医科大学教学医院福建省肿瘤医院肿瘤免疫学研究室,福建福州350014
- 3. 福建医科大学检验科,福建福州350014
- 4. 福建医科大学福省肿瘤转化医学重点实验室,福建福州350014
- 折叠
摘要
Abstract
Objective To analyze the effect of Grb2 inhibition by shRNA transfection on the proliferation of colon cancer cells HT29,and to estimate the feasibility of Grb2 as a target for the treatment of colorectal cancer.Methods The Grb2 shRNA was transfected to 293T cells by shRNA lentiviral vector system,5 strains of different sequence Grb2 shRNA lentiviral particles was obtained and infected colon cancer HT29 cells respectively.5 colon cancer HT29 cell lines infected with the Grb2 shRNA lentiviral particles for the experimental group eGFP shRNA slow virus infection colon cancer HT29 cell for the negative control group.MTF was used to determine the cell proliferation situation,the expression of Grb2 mRNA was detected by RT-PCR and Grb2 、P42/44 ERK、phosphorylation P42/44 ERK、serine/threonine protein (Akt)、phosphorylated Akt (P-Akt)、STAT5 and other molecules signaling pathway detected by Western blot.Results Seventy-two hours after infection,HT29/shGrb2-69,HT29/shGrb2-73 cell were suppression in the mRNA and protein levels.After 24h infection,HT29/shGrb2-69 、HT29/shGrb2-73 cell A values were 0.176 ±0.045、0.186 ±0.013,and 48 h for0.347 ±0.048,0.382 ±0.041 Significantly lower than that of the blank control and negative control(P ＜ 0.001),and 72 hours after infection were 0.934 ±0.038、0.983 ±0.205 (compare with the blank control,negative control,P ＜0.01) ; 72 h after infection,phospho-P42/44 ERK,Akt,phospho-Akt were decreased,while the ERK,STAT5 expression were not affected.Conclusions The application of a lentiviral vector system transfected of Grb2 shRNA to HT29 cells inhibites the level of Grb2 mRNA and protein significantly,induces HT29 cell proliferation and down-regulates signal transduction molecules expression.关键词
结直肠癌/Grb2/RNA干扰/慢病毒/信号通路Key words
colorectal cancer/ Grb2 / RNAi / lentivirus / signaling pathway分类
医药卫生引用本文复制引用
陈丽妹,叶韵斌,刘枋,陈慧菁,李洁羽,苏光建,林巧燕..生长因子受体结合蛋白-2抑制肠癌细胞HT29增殖[J].基础医学与临床,2013,33(9):1199-1204,6.
