RNA Transfection With H2AX And STAT1 Gene In ECA109 Cell Line For Regulation The Cell Cycle And Irradiated Sensitivity

jObjective: To explore the effect on irradiated sensitivity due to expression of protein changed by RNA transfection H2AX and STAT1 gene in esophageal squamous cell carcinoma ECA109 cell lines.Methods:â‘ To observed the effect on proliferation ability of ECA109 cell line with knockdown of H2AX and STAT1 gene by MTT assay.The irradiated sensitivity of ECA109 was checked by clone conformal assay.â‘¡Redistribution of cell cycle and apoptosis index were measured with flow cytometry at 1h,2h,12h,24h and 48h after irradiated by 4Gy.â‘¢The nuclear foci formation of H2AX and STAT1 protein were detected with Zeiss fluorescence microscope after 30 minutes irradiated with RNA transfected cell line.Result:â‘ The proliferation ability of ECA109 cell line was not significantly influenced after RNA transfected of H2AX and STAT1 gene,and p>0.05.â‘¡The cloning formation results of ECA109 showed that the D0 value were 2.98Gy,3.02 Gy,1.96 Gy,2.03Gy and SF2 were 0.88,0.88,0.59,0.83 in ECA109,ECA109-N,ECA109-H,ECA109-S cells respectively.â‘¢The distribution of cell cycle in G₀/G₁,G₂/M and S stage did not change at 12h for none irradiated in ECA109,ECA109-N,ECA109-H and ECA109-S. But the stage of G₀/G₁ were significantly decreased after irradiated by 4Gy. Percentage of G₀/G₁ in ECA109-H,ECA109-S cells line was lower than those in ECA109-H,ECA109-S with no irradiated groups, but it was higher than those in ECA109,ECA109-N irradiated by 4Gy.While retardance of G₂/M were observed at 12h after irradiated by 4Gy.The degree of retardance of G₂/M was more serious in ECA109-H and ECA109-S cells than in ECA109 and ECA109-N cells. Percentage of S stage were significantly decreased at 12h after irradiated by 4Gy. Apoptosis were not influenced at 12h after irradiated by 4Gy. We have obtained similar results at 24h,48h after irradiated by 4Gy with the transfection H2AX and STAT1 gene in esophageal spuamous cell.â‘£After knocking down expression of H2AX and STAT1 gene, the number of H2AX protein nuclear foci was significant decreased and the number of STAT1 nuclear foci was not seen change in ECA109-H cells.Conclusion: H2AX and STAT1 shRNA transfection do not influence proliferation ability of ECA109 cell. Radiosensitivity may be influenced by modulating cell cycle after irradiation.