The Effect Of Apoptosis Migration And Cell Cycle Distribution By Changed Expression Of BMI1 And RNF2 Gene In ECA 109 Esophageal Carcinoma Cell Line

Objective:We investigated the time and dose-effect in protein expression of BMI1 and RNF2 gene by si RNA method after irradiated and explored the influence extend of BMI1 and RNF2 si RNA to the cell proliferation,apoptosis,migration and cell cycle of ECA109 cell of human esophageal carcinoma,in order to supply the foundation for sensitivity of radiotherapy.Methods:1 The level of mRNA and protein expression of BMI1 and RNF2 were determined using reverse transcriptase-polymerase chain reaction(RT-PCR) and flow cytometry(FCM),respectively.The cell proliferation ability was measured by MTT assay method.2 Small interfering RNA(si RNA) which target BMI1 and RNF2 were transferred into ECA109 cells.Distribution of cell cycle and cell apoptosis index were analyzed by flow cytometry,cell migration was detected using Transwell chamber model,and protein expression levels of BMI1 and RNF2 gene were determined using Western blotting analysis.Results:1 The proliferation and cell migration of ECA109 cell1.1 Checking results showed a certain amount m RNA and protein levels expression of BMI1 and RNF2 in ECA109 cells.It was found that the amount of BMI1 and RNF2 protein expression appearing in dose-effect,the greater the dose,the higher the amount of protein expression.1.2 There was a significantly decreased of cell proliferation rate after irradiated at different time comparing with those without irradiated cells.Contrasted with ECA109 and ECA109-N group,the cell proliferationrates of ECA109-B and ECA109-R group were dramatically decreased at 4-24 h by 6Gy irradiated given.1.3 The protein expression levels of BMI1 and RNF2 gene were significantly inhibited when adopting si RNA treated BMI1 and RNF2 gene in ECA109 cells.1.4 The results of cell invasion and cell migration assays indicated: ECA109-B and ECA109-R group cells moved from the upper chamber into the lower one were clearly less than that ECA109 and ECA109-N group cells. 2 The cell cycle distribution and apoptosis of ECA109 cell line2.1 The distribution of cell cycle in G0/G1,G2/M and S stage were not different with none irradiated in ECA109,ECA109-N,ECA109-B and ECA109-R.But retardance of G2/M were observed by irradiated 6 Gy,and stage G2/M were significantly increased after irradiated 6Gy.percentage of stage G2/M in ECA109-B and ECA109-R cells line with irradiated were higher than those ECA109-B and ECA109-R no receiving the irradiated groups,but they were lower than those in ECA109,ECA109-N irradiated 6 Gy.The degree of retartdance stage G2/M were more seriously in ECA109 and ECA109-N cells than that in ECA109-B and ECA109-R cells.There was not influenced of S stage cell in this study.2.2 Cell apoptosis rate significantly increased by irradiated than those in ECA109 with no irradiated.Contrasted with ECA109 and ECA109-N group,ECA109-B and ECA109-R group owned the higher cell apoptosis rates.Conclusion:1 protein expression of BMI1 and RNF2 genes exist in ECA109 esophageal carcinoma cell line.2 Cell proliferation activity and cell migration ability were inhibited,and removed the retardance of stage G2/M and increased the cell apoptosis when BMI1 and RNF2 gene expression were down regulation by si RNA method.