In the present study, Bio-CS significantly promoted the transfection of GM-CSF, IL-21 and Rae-1 in tumor cells, whereas the mRNA and protein expression levels of GM-CSF, IL-21 and Rae-1 in the spleen tissue decreased compared with those noted in the liver cancer tissues that included Bio-CS/plasmid DNA and plasmid DNA. The gene discussed is CSF2; the disease is liver cancer.