Activation Of Expression In Peripheral Blood Mononuclear Cells In Sonic Hedgehog Pathway

Objective :To Discuss the expression and impact of Sonic hedgehog(Shh) blocking antibody(Shh Ab) in peripheral blood mononuclear cells(PBMCs) MC of anti-gastric cancer cells.Methods :Recovery GES-1 cells,cell growth to be 70 to 80%for induction,DMEM 5ml and diluted in serum-free and join the anti-double for the 2×10-1mol/L of MNNG,dark cultured24 h,replacing the serum-containing and double antibody DMEM,7 d about not shedding a lot of adherent cells,the remaining adherent cells were cultured,after digestion and covered passages,this time the cells MC cells,Normal human PBMCs were centrifugated by Ficoll density gradient,(GES-1 cells were treated by nitrite amides)and gastric cancer cells were in co-culture with MC;To observe the expression and semi-quantitative analysis of Shh and Gli-1 by RT-PCR.Shh blocking antibodies were added in the co-culture system,The expression of CD3,CD5,CD69 was assayed by flow cytometry.Results :Significant changes in cell morphology,inverted microscope was polygonal sizes,multicore-shaped cells,cell body larger,tightly packed and lumps growth;RT-PCR:Gli-1m RNA in MC+Shh Ab cell group is 0.2845±0.0025,lower than the MC cell group 0.5167±0.0109(P<0.05),GES-1 gene expression in cells group negative;Flow cytometry:SGES-1 cells group CD3+small cells,MNNG induced group and MC cells group CD3+cells increased significantly,GES-1 cells group and MNNG inducedgroup showed no CD69+cells,MC group of cells CD69+cells was significantly increased,GES-1 a cell group,MNNG-induced cell group and the MC group CD5+cell counts are at a low level.Shh blocking antibody enhanced cell killing PBMCs for MC.Conclusion :Shh blocking antibody can promote activation of PBMCs,and enhanceits anti-carcinogenic effect of nitrite amides.