Abstract:Objective To study the influence of human endostatin 30 peptide modified RGD on the cell adhesion and invasion of HepG2 cells and the mechanism. Methods The nucleotide sequence encoding amino acids 1-30 of endostatin (peptide 30, with amino acids 25-31 mutated from RGIRGAD to RGDRGD) was artificially synthesized and cloned into the plasmid pTYB2 and expressed in E. coli (DE3), Similarly, peptide 27, corresponding to amino acids 1-27 of endostatin, was produced as a control. The cell adhesion assay and transwell was used to inspect the influence of cell adhesion and invasion activity of HepG2 caused by 30 peptide and 27 peptide, respectively. Meanwhile, integrin associate with invasion was screened out; The aggregation of integrin αvβ3 was detected using by immunofluorescence The expression of integrin on the HepG2 cell surface was measured by flow cytometry in order to test the effect of 30 peptide and 27 peptide. The expression of mRNA and protein was tested by RT-PCR and Western blot, including TIMP1 and TIMP2. Results The cell adhesion and invasion of HepG2 was inhibited obviously by 30 peptide, and it's influence of invasive effect on HepG2 associate with integrin αvβ3. At the same time, the mRNA and protein expression of TIMP1 and TIMP2 were down-regulated by 30 peptide. Above function enhanced obviously after αvβ3 antibody joined to 30 peptide. Conclusions 30 peptide can give full play to its anti-transferance for tumor through integrin αvβ3, 30 peptide may help chemotherapeutics to play their function of anti-hepatoma in the clinical treatment.