Construction and functional verification of AP-2 gene family eukaryotic expression vector

Abstract

Objective To construct the eukaryotic expression recombinant plasmids of activator protein 2(AP-2) family genes and to identify the effect of AP-2α, AP-2β and AP-2γ on proliferation and migration of HepG2 cell line. Methods Ap-2α,AP-2β and AP-2γ gene were amplified with human mammary cDNA library as template and ligated with pCDNA3.0-FLAG vector, respectively. The eukaryotic expression vector pCDNA3.0-FLAG-AP-2α/β/γ was constructed and transfected into HepG2 cells. CCK8 assay and clone formation assay were used to detect the effect of AP-2 family genes on HepG2 cell proliferation. Scratch assay was used to detect the effect of AP-2 family genes on HepG2 cell migration. Results The pCDNA3.0-FLAG-AP-2α/β/γ were successfully constructed and expressed in HepG2 cells.AP-2 family genes significantly inhibited the proliferation and migration of HepG2 cells(P＜0.05). Conclusions Ap-2α, AP-2β and AP-2γ may inhibit the proliferation and migration of HepG2 cells.

Keywords

• human ap-2 family genes|hepatocellular carcinoma|eukaryotic expression|cell line hepg2