Study Of Photodynamic Therapy Combined With Adramycin On Proliferation And Cell Cycle Of MCF-7 Breast Cancer Cell Line

Objective:The experiment will study the photodynamic therapy combined with Adriamycin on cell cycle and cell proliferation of MCF7 breast cancer cell line.Methods:The experiment is divided into six phases.The first stage,MTT colorimetric to screen Adriamycin.The second phase,measured the spectra of adramycin and CPD4,meanwhile measured the absorption spectra of the mixture. The third stage,the dye exclusion method to observe the inhibition of the rate of cells.The forth stage,the application of flow cytometry analysed the inhibitory rate of cells in each group.The fifth stage,flow cytometry analysed the cell cycle of each group.The sixth stage,flow cytometry measure the mean fluorescence intensity;Results:1,ADM could inhibit MCF7 cells and the rate of inhibition shows time-concentration -dependent effect;Three concentrations were chosen.Low concentration group was 10ng/ml,20ng/ml,inhibition of MCF-7 cell rate＜10%,high concentrations of 50ng/ml on MCF7 cell inhibition rate＜25%.2,The fluorescence spectra of mixed solution showed blue-shift occurred.It indicted that ADM may combine with CPD4,then in the experiment it avoided given ADM and CPD4 at the same time.3,0.2%eosin exclusion method was used to observe cell rate of death under inverted microscope,the rate showed the ADM can enhance CPD4 photodynamic effect.4,Flow cytometry analysis of cell inhibitory rate showed the ADM can increase the effectiveness of photodynamic effect obviously.5,ADM mainly block cell at G₂/M phase.PDT block cell at G₀/G₁.United drug group mainly block at G₂/M phase.6,The amount of CPD4 entered into cells which were be pretreated by ADM with 24 hours is more than cell.In CPD4 incubated two hours,both reached peak fluorescence intensity and at this time to give light then the rates of inhibition on cell would most obvious.ADM pretreatment group unspent CDP4 incubated two hours after the stable fluorescence intensity;ADM pretreatment group 24 hours after CPD4 incubated two hours with the time to quickly lower the fluorescence intensity. Conclusions:1,Three concentrations of ADM were chosen,10ng/ml,20ng/ml,50ng/ml.2,Selected wavelength of 670nm to excite ADM could not have photodynamic.ADM may combine with photosensitizer then at experiment we should avoid administration of ADM and CPD4 simultaneously.3,Dye exclusion method analysed of cell inhibition rate in each group showed that pretreatment of cells ADM can enhance photodynamic effect,and did not increase toxicity CPD4 dark and light toxicity.4,Flow cytometry analysed of cell inhibition rate in each group showed that pretreatment of cells ADM can enhance photodynamic effect,and did not increase toxicity CPD4 dark and light toxicity.5,ADM,PDT could block cell at different phase of cell cycle.6,CPD4 entered cell more when ADM pretreated MCF-7.Provide a theoretical foundation for ADM can enhance the photodynamic effect.