Experiment Research Of The Proteomic Fingerprint Characteristic Of Esophageal Squamous Carcinoma Cells(ECa109)-Radioresistance

Objective:Study the proteomic fingerprint characteristic of ECa109-IR which support foundation for clinic reaserch. Methods:One.Confirm the sublethal ionizing radiation dose of ECa109. Two. To establish a radioresistant subclone cell line-ECa109-IR. ECa109-IR derived from EC cell line ECa109 by treating the cells with five rounds of sublethal ionizing radiation. Three. Detecting the radiosensitivity of ECa109-IR: using the clonal survival test, cell proliferation assay and flow cytometry to compare the radiosensitivity of ECa109-IR and ECa109. Four.Differences in protein spectrum-- to detecte the protein spectrum of radioresistance ECa109-IR and radiosensitivity ECa109 using protein fingerprint technique.Results: First. Established a radioresistant subclone cell line-ECa109-IR, which derived from EC cell line ECa109 by treating the cells with five rounds of sublethal ionizing radiation. Second. Analysis the radioresistance of ECa109-IR. a. The results of clonal survival test were the cloning of ECa109-IR cells were greater the control ECa109 and the performance is radiation resistance; ECa109-IR dose survival curve downward trend slower than ECa109. Which were proved that the radiosensitivity of ECa109-IR was significantly lower than that in ECa109 and the radioresistance of ECa109-IR was stronger. b. The results of cell proliferation assay show that the effect of radiation on the growth of ECa109-IR was smaller and the recovery time is shorter and the cells growth faster.The results of cell proliferation assay show that the effect of radiation on the growth of ECa109-IR was smaller and the recovery time is shorter and the cells growth faster. c. The flow cytometry show that the percent of G0-G1 between ECa109-IR and ECa109 were no difference, but the S phase of ECa109-IR was more and the G0-G1 was smaller. It illustrateed that the radiation induced G1-S and G2-M phase arrest effect in ECa109-IR was significantly weaker than that in ECa109, which was consistent with the phenotype of the typical radiation resistance. The cell cycle of the radioresistant ECa109-IR has changed. Third. The protein fingerprint of ECa109-IR and ECa109. There are obvious differences in ECa109-IR and ECa109 protein M/Z peak(the peak is 21 when p<0.05,and the peak is 19 when p<0.01). The M/Z were almost less than 6000, and more than half between2000 and 6000. The above show that the protein fingerprint between ECa109 and ECa109-IR was significant difference, in other words, there are different protein between ECa109 and ECa109-IR, and the protein may decided the reason why there have different radiosentivity in ECa109 and ECa109-IR. Conclusion: There are differences protein M/Z peaks between radioresistant ECa109-IR and radiosensitive ECa109, and it is the reason why ECa109-IR radiation resistance, and also provide basis for clinical study.