The Effect Of Different Activated Macrophages On The Growth And Invasion Of Hepatoma Carcinoma Cell

Objective To explore the phenotypic characteristics of differentactivated macrophages in vitro culture and their effect on the growthand invasion of Hep G2liver cancer cells and provide theory evidencefor the macrophage cells in the liver cancer immunotherapy.Methods Mononuclear cells were isolated from the peripheralblood of healthy adults and differentiate into macrophages byGM-CSF in vitro culture. Classic activation of macrophage (M1orTAM1) was induced by LPS and IFN-γ; Alternative activation ofmacrophage (M2or TAM2) was induced by IL-4. The CD68andCD206expression in three different activated macrophage weredetected By flow cytometry; IL-10and IL-12secretion levels weredetected by ELISA; Relative expression of mRNAs of TNF-alpha,CCL3, iNOS, Arg-1and AMAC-1were detected by RT-PCR; Thegrowth inhibition rate and invasion effect of different activatedmacrophages on Hep G2liver cancer cells was detected by MTTmethod and Transwell invasion assay respectively.Results1. The phenotypic characteristics of different activatedmacrophages4) UaM, TAM1and TAM2have a high expression of CD68and the expression rate was42.52±4.52%,61.53±5.29%and66.28±6.87%respectively; The CD206expression rates were3.61±0.84%,0.86±0.46%and32.16±3.61%respectively, TAM2CD206expression rate was significantly higher than UaM and TAM1(P<0.05).5) There was no statistically significant difference between UaMand TAM1IL-10secretion (P>0.05), and both of them IL-10secretionwere lower than TAM2(P <0.05); IL-12secretion in UaM, TAM1andTAM2did not exist statistical difference (P>0.05). TAM1had thegreatest secretion of IL-12and UaM had the fewest secretion of IL-12.6) The expression level of TNF-α, CCL3and iNOS mRNA inTAM1was higher than in TAM2(P <0.05). But TAM1had a lowerexpression level of AMAC-1and Arg-1mRNA than TAM2(P <0.05).2. The effect of different activated macrophages on the growthand invasion of hepatoma carcinoma cell3) TAM1culture supernatant had a significantly higher growthinhibition rate on Hep G2cells than UaM and TAM2(P <0.05). Theincresing of growth inhibition rate on Hep G2cells was associatedhigher concentration of TAM1culture supernatant. However, theincresing of growth inhibition rate on Hep G2cells was associatedlower concentration of TAM2culture supernatant.4) The number of cells passing through basement membrane inthree groups had statistically significant difference. TAM1+Hep G2had the greatest number of cells passing through basement membraneand TAM2+Hep G2had the leatest number of cells passing throughbasement membrane. Conclusion：1. TM1and TAM2, induced in vitro, are two types ofmacrophage subsets with different phenotypes.2. In vitro, TAM1can effectively inhibit the proliferation andinvasion of Hep G2hepatoma cells and play roles in antitumor effectand TAM2promote the proliferation and invasion of Hep G2hepatoma cells and play roles in promoting tumor progression.