Green fluorescent protein reporter gene labeling humans prostate cancer PC-3 cells mediated by lentivirus

Objective To explore the effect of green fluorescence protein(GFP)labeling mediated by lentivirus on the biophysical properties of humans prostate cancer PC-3,and whether the GFP gene expression is permanent and stable,which can lay the foundation for further experiments.Methods Conventional tumor cell culture,passage.Multiplicity of infection(MOI) of different viruses in the cell state was the best implementation of GFP lentivirus infection of PC-3,and 7 days under a fluorescence microscope to observe GFP expression in PC-3.Selected the infection with GFP-positive expression rate of the highest hole to continue fostering the passage to the third generation,respectively with the microscope.Morphology and MTT measured growth curve,the scratch test to compare the two cell morphology,activity,growth rate and growth of state,to evaluate the GFP the stability of gene expression in PC-3 cells.Results Under the fluorescence microscope,72 hours later,green fluorescent was found and became strongest at 7 days.Among them with MOI=20 highest infection rate,the GFP positive expression rate was(92.3±1.2)%.GFP / PC-3 to sustain the culture in vitro,2,4,8 weeks of GFP-positive rate did not change significantly.Microscopically form view,MTT measured growth curve,scratches experimental results show that compared with before transfection,lentiviral infection after PC-3 cell viability,proliferation,apoptosis and cell cycle did not affect(P0.05).Conclusions GFP lentivirus can efficiently label PC-3 cells and has no significant effect on the biophysical properties of PC-3 cells.GFP gene expression in PC-3 cells is permanent and stable.GFP / PC-3 can be used for further cell tracing research.