Radiosensitization Effect Of Recombinant Adenovirus-p53on Pancreatic Carcinoma Cell Line Aspc-1

Objective: To evaluate the sensitization effect and mechanisms of recombinanthuman adenovirus p53injection on the pancreatic cancer cell ASPC-1；Explore therelationship and interaction among p53, p21and Bax of p53pathway related geneexpressing on cell ASPC-1；Estimate apoptosis and radiosensitizing effect on humanpancreatic cancer cell after transfection of recombinant human adenovirus p53（rAd-p53）.Methods: We choose human pancreatic cancer ASPC-1cell for the study in vitro.The experiment was divided into the control group (group C), the irradiation group(group R), drug group (group D), irradiation plus drug group (group D+R). Detectthe radiosensitizing effect on the ASPC-1cells after injection of rAd-p53via cellcolony formation assay, observe cell survival situation and draw the cell survivalcurve, use the clicking multy target teaching model to fit the radiation dose-effectcurve; Analyse the cell cycle distribution and apoptosis index48h after4Gyirradiation by flow cytometry. Detect the expression of p53and related gene p21, baxon cell ASPC-1at different time after p53transfection (24h,48h,72h) throughImmunocytochemistry.Results:1.Colony formation test showed that cell colony-forming abilitydecreased significantly after rAd-p53transfection and different doses of irradiationexposure，along with the increase dose down more obviously；under the same dose,the ability of group D+R cells of the lowest；Aspc-1cell irradiated group (group R)in D0is2.398,while irradiation dosing group (group D+R) in D0is2.166, indicatingthere was radiosensitizing effect on ASPC-1cell by rAd-p53injection.2.The cell cycle results showed that after ASPC-1transfected with rAd-p53, the proportion ofG2/M cell increased, and G2/M phase arrested the most strongest in group (D+R)(P <0.05). Meanwhile the apoptosis index was higher in group (D+R). G2/M cellsand apoptotic index correlation analysis showed that there was a positive relationshipeach other mutually with（rs=0.917, P<0.001）.3.The immunocytochemistry testresults showed the protein expression of Bax, p21, p53in drug group were more thanthose of control group, and the level was different at different time after p53transfection in each group.It was the most after72hours.Conclusions: The rAd-p53could transfect into pancreatic cancer cell ASPC-1,and show significant radiosensitization biological effects. The study preliminarilyelucidate the mechanism of rAd-p53in pancreatic cancer cells that providesexperimental basis for clinical study.