Construction of lentiviral vector pLeno-DCE-PDGF-B and its effect on invasion ability of SGC7 9 0 1 gastric carcinoma cells / 重庆医学

ABSTRACT

Objective To construct a lentiviral vector that stably express platelet-derived growth factor B (PDGF-B)and to in-vestigate effect of PDGF-B overexpression on invasion of SGC7901 gastric carcinoma cell.Methods The lentiviral vector pLeno-DCE-PDGF-B was constructed and transfected into 293T cells.The supernatant containing the lentivirus particles was harvested to determine the virus titer and high titer lentivirus particles was gathered.Then PDGF-B lentiviral vector was transfected into SGC7901 cells for construction of stable PDGF-B overexpression SGC7901 cells.Western-blot and immunofluorescence were used for evaluation of the construction of stable PDGF-B overexpression SGC7901 cells.Transwell test was used for detection the inva-sion of SGC7901 cells.Results The lentiviral vector was correctly constructed and verified by sequencing.High titer PDGF-B lenti-viral particles were acquired successfully.After transfection,SGC7901 cells were green color by detection of fluorescence micro-scope;and the expression of PDGF-B protein in transfected SGC7901 cells were much higher than that in normal SGC7901 cells.Al-so,PDGF-B overexpression increased the invasion ability of SGC7901 cells.Conclusion The PDGF-B lentiviral particles are ob-tained after successful construction and package.The stable PDGF-B overexpression SGC7901 gastric carcinoma cell line is estab-lished,which proves that the PDGF-B overexpression might increase the invasion ability of gastric carcinoma cells.