Effect of target down regulation of epithelial cell adhesion molecule on the function and drug sensitivity of colorectal cancer stem cells
10.13699/j.cnki.1001-6821.2018.01.009
- VernacularTitle:靶向下调上皮细胞黏附分子对结直肠癌干细胞功能及药物敏感性的影响
- Author:
Xue-Song TU
1
;
Li-Xia HU
;
Guang-Qiao QU
;
Hui-Ting XU
Author Information
1. 孝感市中心医院药学部
- Keywords:
colorectal cancer stem cell;
epithelial cell adhesion molecule;
cluster of differentiation 44;
proliferation;
invasion;
drug sensitivity
- From:
The Chinese Journal of Clinical Pharmacology
2018;34(1):34-37
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of epithelial cell adhesion molecule(EPCM) expression downregulation on the proliferation,invasion and drug sensitivity of ECAM+/cluster of differentiation 44 (CD44) + colorectal cancer stem cells.Methods Colorectal cancer stem cells (ECAM) were obtained from human colorectal cancer cell line LoVo by the method of tumor microspheres.The specific markers of colorectal cancer stem cells were identified and studied.Lipofectamine 2000 liposome was used to complete the transfection,which was divided into three groups:blank group (ECAMhjgh/CD44 + LoVo),control group (transfection of common inhibitors,lipofectamine 2000-inhibitors) and experimental group (ECAM) according to different treatment methods.They were cultured in the same way,the three group of cells in logarithmic phase were treated with irinotecan (drug concentrations were 1,5,10,15,20,25,30,35,40,45,50 mg · L-1) and capecitabine (drug concentrations were 50,100,150,200,250,300,350,400,450,500,550,600mg· L-1).Then,they continued to be cultured.The expression levels of ECAM mRNA in three groups of ceils were detected by real-time fluorescence quantitative detection (RT-PCR).The thiazolyl blue tetrazolium bromide (MTI)assay was used to detect the proliferation and drug sensitivity of the three groups before and after treatment with irinotecan and capecitabine.Transwell cells were used to detect the invasion ability of three groups of cells.Results The percentage of ECAMhjgh/CD44 + colorectal cancer stem cell in LoVo cell line was 0.95％,85.78％ before and after enrichment,and the difference was significant (P ＜ 0.05).The expression of ECAM mRNA in blank group,control group and experimental group were 8.17 ±0.64,7.94 ±0.83,2.16 ±0.12.Compared with blank group,the difference had significantly in two groups (P ＜ 0.05,P ＜ 0.01).Compared with control group,the difference in experimental group had significantly in two groups (P ＜ 0.05).It indicated that model was succeed prepared.Invasive cell in the three groups were 79.22 ± 5.25,80.12 ± 4.89,31.23 ± 2.36.Compared with blank group,the difference had significantly in two groups (P ＜0.05,P ＜0.01).Compared with control group,the difference in experimental group had significantly(P ＜0.05).The IC50of irinotecan on colorectal cancer stem cells in the three groups were (20.25 ±4.35),(19.22 ±3.99),(10.24 ± 2.04) mg · L-1.The IC50of capecitabine on colorectal cancer stem cells in the three groups were (320.13 ± 23.65),(315.79 ± 21.03),(250.22 ± 15.45) mg · L-1.Compared with control group,the difference in experimental group had significantly(P ＜0.05).Conclusion Targeted down-regulation of ECAM can effectively inhibit the proliferation and invasion of colon cancer stem cells,and enhance their sensitivity to drugs at the same time.
