Construction of prokaryotic expression vector with human esophageal cancer related gene 4 and the effect of recombinant protein on the cell cycle of EC-18 in esophageal carcinoma
10.3760/cma.j.issn.1006-9801.2018.11.001
- VernacularTitle:人食管癌相关基因4原核表达载体构建及其重组蛋白对食管癌EC-18细胞周期的影响
- Author:
Linwei LI
1
;
Wenyu WANG
;
Xiaoyan LI
;
Zuxuan SHI
;
Tianhui GAO
Author Information
1. 河南省人民医院 河南大学人民医院 郑州大学人民医院肿瘤科
- Keywords:
Esophageal cancer related gene 4;
Recombinant protein;
Cell cycle
- From:
Cancer Research and Clinic
2018;30(11):721-724
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the prokaryotic expression vector of human esophageal cancer related gene 4 (ECRG4), to purify the recombinant ECRG4 protein and to verify the biological function of the recombinant ECRG4 protein. Methods DNA recombination technology was utilized to construct the ECRG4 protein prokaryotic expression vector. The recombinant ECRG4 protein was purified with the transformation of escherichia coli. Then the purity of the recombinant ECRG4 protein was examined by using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Furthermore, esophageal cancer EC-18 cell line was treated by recombinant ECRG4 protein (10 μg/ml) for phosphate buffer (PBS) 48 h respectively, and the tumor cell cycle change was examined by using flow cytometry. Results SDS-PAGE analysis showed that the high purity of recombinant ECRG4 protein was obtained and the ECRG4 protein prokaryotic expression vector was successfully constructed. The cell proportion of G1 phase in PBS control group was lower than that in the recombinant ECRG4 protein group [(60.4 ±2.1) ％ vs. (71.6 ±1.8) ％; t= 25.695, P= 0.002]. The cell proportion of S phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(24.6±1.4) ％ vs. (16.5±1.0) ％; t= 36.905, P= 0.001]. The cell proportion of G2/M phase in PBS control group was higher than that in the recombinant ECRG4 protein group [(15.0 ±1.1) ％ vs. (11.9 ±0.8) ％; t=6.471, P=0.023], which indicated that the recombinant ECRG4 protein could induce the G1 phase arrest of EC-18 cells. Conclusion The ECRG4 protein prokaryotic expression vector is successfully constructed. And the recombinant ECRG4 protein has an active biological function in esophageal carcinoma.
