Objective:To construct recombinant glucose regulated protein(GRP78)-expressed adenovirus

to establish a cell model by infecting HepG2 cells

and to study the effect of recombinant GRP78 adenovirus on cell proliferation in the HepG2 cells. Methods:Plasmid pAd Track-TO4-GRP78 containing GRP78 gene was constructed by using adenovirus shuttle plasmid pAd Track-TO4

and the sequence was identified by restriction enzyme digestion method. After linearized by Enzyme digestion of PmeⅠ

it was homologous recombination with Ad Easy-BJ5183 cells. The recombinant adenovirus vector was linearized by PacⅠ enzyme and transfected into HEK293 cell. The packaging of recombinant adenovirus was judged by the expression of the enhanced green fluorescent protein.GRP78 recombinant adenovirus was extracted by repeated centrifugation and low temperature cracking. HepG2 cells were infected with recombinant adenovirus. The expression of GRP78 gene was detected by qRT-PCR. The expression of GRP78 protein was detected by Western blot. Then the GRP78 adenovirus infected HepG2 cells. The effect of recombinant GRP78 adenovirus on cell proliferation was detected by MTS colorimetric assay.Results:It was proved by gene sequencing that pAd Track-TO4-GRP78 was successfully constructed. And by enzyme digestion validation

the recombinant adenovirus vector pAd-GRP78 was successfully constructed. After pAd-GRP78 being transfected into HEK293 cells

the GPF fluorescence results indicated that the virus was successfully packaged. HepG2 cells were infected with recombinant virus. And the expression of GRP78 gene was confirmed by qRTPCR

and the expression of GRP78 protein was confirmed by Western blot. MTS showed that the recombinant expression of GRP78 adenovirus could promote the proliferation of HCC cells. Conclusion:GRP78 recombinant adenovirus vector was successfully con structed

and the recombinant adenovirus vector could infect HepG2 cells and express GRP78 protein. GRP78 adenovirus could promote the proliferation of HCC cells. This study lay a foundation for further research on the function of GRP78.