M1and M2Tumor-associated Macrophages Affect The Cell Growth, Apoptosis And VEGF-C Protein Expression Of Gallbladder Cancer

Objective: To investigate M1and M2tumor-associated macrophages affect thecell growth, apoptosis and VEGF-C protein expression of gallbladder cancer in vitro.Methods: Monocytic THP-1cells were induced into macrophases byphorbolmyristate acetate（PMA）, then differentiate into two subgroups: interferon-γ（IFN-γ）and Lipopolysaccharid（LPS）used to obtain M1macrophases, and interleukin-4（IL-4）used to obtain M2macrophases. Then their phenotypes were identified byCD80and CD163using analysis of flow cytometry. These two subgroups andmonocytic THP-1cells were cultured with gallbladder carcinoma cells by transwellsystem, divided into four groups: group M1, group M2, group monocytic and controlgroup. The number of living gallbladder carcinoma cells of these groups were countedand establishing the growth curve by CCK method. Used flow cytometry instrumentdetected the ability to induce apoptosis of the gallbladder carcinoma cells. Thechanges of vascular endothecial growth factorC（VEGF-C） of the gallbladdercarcinoma cells from different groups were examed by using Western blottinganalysis.Resμlts: Monocytic THP-1cells induced by IFN-γ/LPS, converting M1macrophages express positive rate of CD80is68.83±2.77%(p<0.01), and induced byIL-4converting M2macrophages express the positive rate of CD163is73.88±3.95%(p<0.01). The group M2gallbladder carcinoma cells grew faster than thegroup M1，group THP-1and control group, the differences were remarkabl（ep<0.01）.The group M1gallbladder carcinoma cells grew slower than the group M2，groupTHP-1and control group, the differences were remarkable （p<0.01）. The apoptosisrate of group M1gallbladder carcinoma cells were higher than control group, groupTHP-1and group M2（p<0.01）, while group M2compared with the control group,group THP-1showed no statistically significant （P>0.01）. The level of VEGF-Cprotein of the gallbladder carcinoma cells fromgroup M1markedly lower than control group, group THP-1（p<0.05）, with the increase of time elapsed and decreases. Andin this case, the group M2got the opposite result, up-regulatingthe expression of VEGF-C were higher than the control group and THP-1group atthe corresponding time（p<0.05）, increased ad the time went on.Conclusion:（1）M1macrophages could promote the GBC-NOZ cells apoptosis,inhibition their growth, down-regulation the expression of VEGF-C protein.（2）M2macrophages could promote the GBC-NOZ cells growth, up-regulation the expressionof VEGF-C protein, but no affect on the apoptosis of the GBC-NOZ cells.