Poorly Differentiated And Undifferentiated Gastric Cancer Cell-derived Microparticles Can Influnce High-differentiated Gastric Cancer Cell's Biological Characteristics

Objective The aim of this study is to investigate the potential influences about aggressive phenotype(mesenchymal properties?migration ability?invision ability and proliferation ability)of poorly-differentiated and undifferentiated gastric cancer cell-derived microparticles on the high-differentiated gastric cancer cells.Methods1.The expression and location of E-cadherin and vimentin in three gastric cancer cell lines(NCI-N87 cell,AGS cell,HGC-27 cell)were examined by immuno-histochemical staining.2.Microparticles were obtained by hypothermal differential centrifugation.The size and its distribution of microparticles was detected by NTA.FCM was used to detect the size and identify the immuno-phenotype of the microparticles.3.The expression of E-cadherin and vimentin in three gastric cancer cell lines(NCI-N87 cell,AGS cell,HGC-27 cell)and NCI-N87 cells treated by AMPs and HMPs were detected by Western-Blotting analysis and q RT-PCR analysis.4.Transwell analysis was used to detect the migration ability and invision ability of three gastric cancer cell lines(NCI-N87 cell,AGS cell,HGC-27 cell)and NCI-N87 cells treated by AMPs and HMPs.5.Imunno-inflourescence technique was used to evaluate the expression of f-actin protein of NCI-N87 cells dealed with AMPs and HMPs.Western Blotting assay was used to detect the expression level of MMP-9 protein of NCI-N87 cells treated by AMPs and HMPs.6.MTS method was used to detect the proliferation ability of AMPs and HMPs treated NCI-N87 cells and three gastric cancer cell lines(NCI-N87 cell,AGS cell,HGC-27 cell).Results1.Poorly-differentiated AGS cells and undifferentiated HGC-27 cells comparision to high-diffentiated NCI-N87 cells,the exprssion of E-cadherin was obviously reduced,the exprssion level of vimentin was signifigantly elevated(P<0.05).2..Poorly-differentiated AGS cells and undifferentiated HGC-27 cells comparision to high-diffentiated show more migraton ability?invision capicity ?proliferation ability(P<0.05).3.NTA ? FCM were used to detect the microvesicles whose diameter is 0.1-1?M.Annexin?antibody and CD326 antibody were used to marked the microparticles,the double positive percentage of AMPs ? HMPs were respectively 94.2% ? 93.5%,which implies that the microparticles we abtained are derived from AGS cells and HGC-27 cells.4.HMPs can induce high-differentiated NCI-N87 cells to express less E-cadherin and more vimentin in both protein level and m RNA level,to show more migration ability and prolifereation capicity(P<0.05).However,invision ability of NCI-N87 cells has not significantly been enhanced after treated by HMPs(P>0.05).AMPs has no significant influnce on the expression of E-cadherin and vimentin,migration ability,invision ability and prolifereation capicity of NCI-N87 cells(P>0.05).5.The f-actin skeleton of NCI-N87 cells treated by HMPs increased(P<0.05).The expression level of MMP-9 protein of NCI-N87 cells treated by HMPs has no significant changes compared with NCI-N87 cells treated by AMPs(P>0.05).The f-actin skeleton and MMP-9 protein of NCI-N87 cells treated by AMPs has no significant changes(P>0.05).Conclusion1.Poorly-differentiated and undifferentiated gastric cancer cell show more mesenchymal properties ? migration ability invision ability and profliferation ability in comparison with high-differentiated gastric cancer cell.2.Undifferentiated gastric cancer cell-derived microparticles(HMPs)can induce high-differentiated gastric cancer cells(NCI-N87 cell)to show more mesenchymal properties ?migration ability and profliferation ability.However,poorly differentiated gastric cancer cell-derived microparticles(AMPs)can not induce NCI-N87 cell to show more significant aggressive phenotype.