To investigate the effect of serum containing Zhuang medicine Fuzheng Prescription on proliferation,migration,invasion,and other biological behaviors of lung cancer A549 cells in the co-culture system composed of tumor-associated macrophages(TAMs) and lung cancer A549 cells and its possible mechanism.Methods:Human acute monocytic leukemic cell line(THP-1) was induced by phorbol-12-myristate-13-acetate(PMA) combined with interleukin(IL)-4 and IL-13,to establish the in vitro model of TAMs.Real-time quantitative polymerase chain reaction(RT-PCR) and enzyme-linked immunosorbent assay(ELISA) were used to analyze the phenotypic regulation of TAMs by medicine-containing serum(low,medium,and high-dose groups) and 24,48,and 72 h after administration.By using the supernatant co-culture system and taking lung cancer A549 cells as the research object,methyl thiazolyl tetrazolium(MTT) assay,real-time cell analysis(RTCA) technology,and Transwell assay were used to determine the proliferation,migration,and invasion of A549 cells under different conditions.Results:The serum containing Zhuang medicine Fuzheng Prescription significantly up-regulated the expression of M1-type markers IL-12 and inducible nitric oxide synthase(iNOS),and down-regulated the expression of M2-type markers IL-10 and CD206.As compared with the blank group,the proliferation of A549 cells in the co-culture group was increased,and the migration and invasion of A549 cells were improved(P<0.05).As compared with the co-culture group,the proliferation,migration,and invasion of A549 cells in the low,medium,and high-dose medicine-containing serum groups were inhibited in a dose-dependent manner(P<0.05).Conclusion:The serum containing Zhuang medicine Fuzheng Prescription can inhibit the proliferation,migration,and invasion of lung cancer A549 cells in the co-culture system,and the mechanism may be related to the regulation of the phenotype of TAMs in the tumor microenvironment.