Study Of The Influence Of RhPDCD5 Protein On Chemotherapy In Lung Adenocarcinoma A549 Cells

Objective:The purpose of this study was to investigate the effect of rhPDCD5 combined with cisplatin on chemotherapy of A549 cells and to explore its possible mechanism.Methods:1.Draw the A549 cell growth’s curve by Cell counting method; 2. Using laser scanning confocal fluorescence microscope to detection the expression of PDCD5 protein after treatment with various concentrations of rh PDCD5; 3. Using Elisa method to measure the expression of PDCD5 protein after treatment with various concentrations of cisplatin; 4. The proliferation inhibition rate of the A549 cells that were treated with various concentrations of rh PDCD5 and cisplatin on 24 h were detected by MTT assay; The cells were divided into 5 groups: the control group, rh PDCD5(15 mg/L) group, cisplatin group(20 mg/L), rh PDCD5 15 mg/L + cisplatin(20mg/L) group, rh PDCD5 30 mg/L + cisplatin(20 mg/L) group,after 24 h treatment, the effects of drugs ongrowth were detected by MTT assay; 5. The apoptoses and cell cycle were measured by flow cytometric; 6. The Caspase 3 activity was detected by fluorescence detection kit.Results:1.Draw A549 cell’s survival curve according the cell counting method and the cell doubling time was 18 h; 2. Laser scanning confocal fluorescence microscope showed that with the increase of rh PDCD5 protein concentration, the expression of PDCD5 protein in A549 cells was increased; 3. The ELISA showed that with the increase of cisplatin concentration, the secrete of PDCD5 protein in A549 cells was increased(F = 354.125, P < 0.001); 4. The proliferation inhibition rate of the cancer cells treated with different concentrations of rh PDCD5 were similar to those of controls(F=0.114, P=0.949), while those of the cells treated with different concentrations of cisplatin(F=88.564, P<0.01); Compared to the single treatment, the combined treatment with rh PDCD5 and cisplatin has a higher inhibiting cell proliferation(F = 130.147, P < 0.001), the proliferation inhibition rate of the control group, rh PDCD5(15 mg/L) group, cisplatin group(20 mg/L), rh PDCD5 15 mg/L + cisplatin group 20 mg/L, rh PDCD5 30 mg/L + cisplatin group 20 mg/L were(0.00±0.00)%,(0.6±1.07)%,(49.67±1.43)%,(68.17±1.49)%,(78.03±0.94)%, respectively, and there was statistically significant among these five groups(F=1095.472,P<0.001); 5.The G0 / G1 phase of the groups were(26.43±0.95)%,(27.34±0.34)%,(47.88±0.39)%,(59.75±0.38)%,(73.78±2.11)%, respectively, there was statistically significant among these five groups(F=365.501, P<0.001); 6.Apoptotic ratio of the groups were(6.563±0.49)%,(5.07±0.49)%,(22.31±0.64)%,(43.30±0.62)%、(64.25±0.54)%, respectively, there was statistically significant among these five groups(F=2054.594, P<0.001); 7. Fluorescent protein activity tests showed the groups’ Caspase 3 activation were 22.28±0.06, 2.26±0.13, 3.93±0.25, 9.76±0.11, 11.62±0.24, respectively, there was statistically significant among these four groups(F=877.907, P<0.001).Conclusion:1.RhPDCD5 protein could enter the A549 cells to play a role, and cisplatin could promote the expression of PDCD5 in A549 cells in vitro; 2.Rh PDCD5 protein alone couldn’t work for A549 cells; 3.Rh PDCD5 combined cisplatin could arrest A549 cell line cell cycle in G0 / G1 phase, and inhibit A549 cells proliferation; 4.Rh PDCD5 protein combined cisplatin could enhance the activation of Caspase-3 to promote apoptosis.