论文部分内容阅读
Objective: The aim of the study was to construct the eukaryotic expression vector of human angiopoietin-like protein 4(ANGPTL4)and observe the effect of ANGPTL4 overexpression on the growth of esophageal carcinoma EC9706cells.Methods: Total RNA was extracted from normal hepatic tissue,and ANGPTL4 cDNA was amplified by RT-PCR.The PCR product was doubly digested by Xbal and Sall,and then recombined into eukaryotic expression vector.Then,pIRES-GFP-ANGPTL4 was obtained by G418 selection,then pIRES-GFP-ANGPTL4 and pIRES-GFP were transfected into EC9706 cells with lipidosome-packaged method.Meanwhile,the transfected cells were selected by G418,and then stable transfected cell lines were obtained.ANGPTL4 mRNA levels,the cell cycles and growth curves of EC9706 cells in experiment group(transfected with pIRES-GFP-ANGPTL4),empty vector group(transfected with pIRES-GFP)and blank control group(EC9706 cells without transfection)were detected with RT-PCR,flow cytometry and MTT methods,respectively.Results:Eukaryotic ANGPTL4 expression vector pIRES-GFP-ANGPTL4 was successfully constructed.The ANGPTL4 mRNA level(0.21 ± 0.03)in experiment group was significantly higher than that of the empty vector group(0.04 ± 0.008)and the blank control group(0.05 ± 0.007),with significant differences(P < 0.01).The proportion of cells in S phase in experiment group was significantly different with those of the other two groups(P < 0.05).The cell growth of EC9706 cells in experiment group was slower than those of the other two groups.From the third day,the differences began to be significant.Conclusion:ANGPTL4overexpression in esophageal carcinoma EC9706 cells could inhibit the growth of EC9706 cells.