The Effect Of Human Embryonic Stem Cell On HepG2 And A549 Tumor Cells

Objective h ESCs-HepG2 and hESCs-A549 non-contact co-culture system were established to explore influences of human embryonic stem cells(hESCs)on invasion,migration,hproliferation and apoptosis of HepG2 and A549 cells to provide basis theories for stem cell therapy for cancer.Methods HepG2,A549 were repectively co-cultured with h ESCs to establish h ESCs-HepG2 and hESCs-A549 non-contact co-culture systems by Transwell chamber to provide micro-environment of hESCs for tumor cells.HepG2 and A549 cells which co-cultured with hESCs act as Co-HepG2 experimental group and Co-A549 experimental group,while which respectiely cultured in Transwell act as HepG2 control group and A549 control group.After co-cultured with hESCs,the invasion and migration abilities of HepG2 and A549 cells were detected by Transwell chamber method;the hproliferation ability were measured by MTT method;the effect of hESCs on cell cycle was detected by Flow Cytometry;r nucleus variation and cell apoptosis of HepG2 and A549 were detected by Hoechst33258 chromosome and Flow Cytometry.Finally,qRT-PCR method and Western Blot method were used to detected the expression ofCyclind1,CDK4,P21,Bax and Bcl2 in HepG2 and A549.Results In invasion experiment,the number of cells crossing the Transwell chamber in Co-HepG2 group were less than that in HepG2 control group;transmembrane cells in Co-A549 were less than that in A549 group obviously(P<0.05).In migration experiment,cells crossing the Transwell in Co-HepG2 group were decreased significantly control group;cells in Co-A549 group witch crossing Transwell were decreased compaired with A549 control group(P<0.05).Cell hproliferation experiment shows that the hproliferation ability of cells in Co-HepG2,Co-A549 were both inhibited than their control group obviously(P<0.05),and this inhibitory effect gets increased with time going by;cells in G1 phase of Co-HepG2 were significantly increased compaired with control group(P< 0.05);cells in co-A549 group which were stoped in G1 phase increased significantly than in A549 control group(P<0.05).After co-cultured whith hESCs,the relative expression of CDK4 and CyclinD1 mRNA were less in co-HepG2 group significantly compared with HepG2 group,while P21 were more in co-HepG2 group(P<0.05);Expression of CDK4,CyclinD1 mRNA in co-A549 were decreased and P21 mRNA increased significantly detected by qRT-PCR experiment which were reproved by Western Blot.Hoechst33258 dyeing experiments found that cells whose nuclei got Contraction,deformated and concentrated in co-HepG2,co-A549 group were much more than that in control groups.Cell apoptosis rate inco-HepG2 group increased significantly compaired with HepG2 control group(P<0.05);qRT-PCR and Western Blot experiments show that the expression of Bax increased and the expression of Bcl2 decreased obviously.Conclusion Human embryonic stem cells(h ESC)can inhibit invasion,migration,proliferation and promote apoptosis of HepG2,A549 cells.The experimental results show that hESCs may has inhibitory effect on a wide variety of tumor cells in vitro.