The Effect Of Kanglaite Injection Combined With Gefitinib On The Proliferation And Apoptosis Of Human Lung Adenocarcinoma Cells

Purpose:To investigate the effect of kanglaite injection combined with gefitinib and two drugs on the proliferation of gefitinib resistant H1299 cells in human lung adenocarcinoma and its mechanism.Material and method:1.The human lung adenocarcinoma H1299 cells resistant to gefitinib in logarithmic growth phase were taken and divided into the control group,the KLT group and the gefitinib group and the combination group.The blank control group was added with complete culture medium,and the drug concentration of the KLT group was 5,10,20,50,80,100,160μl/ml;The drug concentration of gefitinib group was 5,10,20,30,40,50μmol/L,The concentration of KLT and gefitinib of IC₅₀0 and KLT（5,10,20,50,80,100,160μl/ml）.CCK-8 method was used to detect the effect of KLT and gefitinib on cell proliferation.2.The logarithmic phase the treatment of human lung adenocarcinoma H1299 cells resistant strains,the treatment group is divided into blank control group,the KLT group,the gefitinib group and two drugs combination group.The control group was added with complete culture medium,the KLT group drug concentrations of 148μl/ml,the gefitinib group drug concentrations of 40μmol/L,the drug concentration of 20μl/ml KLT and the drug concentration of 40μmol/L,the treatment by flow cytometry analysis KLT,gefitinib and combined the two drugs effects on cell apoptosis and cycle;Western Blot was used to detect the expression level of Bcl-2 protein and Bax protein in cells of each experimental group.Results:1.The different concentrations of KLT group and gefitinib group could inhibit the proliferation of H1299 cells（P<0.05）,with an IC₅₀0 value of（143.8±2.74）μl/ml、（40.36±1.34）μmol/L,and the inhibitory rate of KLT group and gefitinib group was significantly higher than that of the KLT group and gefitinib group（P<0.05）.The inhibitory rate of KLT group and gefitinib group was also significantly higher than that of the control group（P<0.05）.2.Flow cytometry was used to detect H1299 cells treated with KLT combined with gefitinib for 48 hours.The apoptosis rate of KLT combined with gefitinib was higher than that of the blank control group and the single drug group,and the difference was statistically significant（P<0.05）.Compared with the blank control group,the proportion of G1 cells in the gefitinib group in KLT group was significantly increased,and the proportion of G1 cells,and the proportion of s and G2 cells was significantly reduced.in the KLT combined with gefitinib group was significantly increased（P<0.05）.3.The expression of Bcl-2 protein in the KLT group and the gefitinib group was lower than that in the blank control group,and the expression of Bax protein was higher than that in the blank control group,with statistically significant differences（P<0.05）.Conclusion:1.Both KLT and gefitinib could inhibit the proliferation of human lung adenocarcinoma resistant strain H1299 cells in a time dependent and concentration dependent manner,and the inhibitory effect was more significant after the combined effect of the two drugs;2.The KLT and gefitinib can synergistically induce apoptosis of human lung adenocarcinoma drug resistant strain H1299 cells,and its mechanism may be related to down regulation of Bcl-2 protein expression and up regulation of Bax protein expression.