Experimental Study Of Berberine Combined With Cisplatin In Inducing Apoptosis Of Human Lung Cancer Cell Line A549/DDP Resistant To Cisplatin

Objectives:Through the experimental observation of berberine in combination with cisplatin resistance to human lung cancer cell line A549 / DDP proliferation and apoptosis and the effect of berberine combine cisplatin induced apoptosis of human lung cancer cell line A549/DDP resistant mechanism.Methods:The study subjects were human lung cancer resistance A549 / DDP cells,each experimental groups were added with 12μg/m L Cisplatin,20μmol/L、40μmol/L、80μmol/L berberine with 12μg/m L Cisplatin,respectively,blank control wells(PBS wells)were set up,they were all treated with drugs for 24 h,48h and 72 h,cell proliferation was evaluated by MTT assay;With drugs for 48 h,cell apoptosis was determined by using the flow cytometry instrument,Real-time PCR and western blotting analysis was measured to detect the levels of cell apoptosis including Caspase-3、Caspase-9、Bax、Bcl-2 in m RNA and protein.Results :(1)The absorbance of the A549 / DDP cells between the group of different concentrations of berberine in combination with cisplatin,cisplatin group and PBS group was significantly different(P < 0.01)at the first 24h;The absorbance was decreased and the inhibition rate was increased with the concentration of berberine increased,but there was no statistical difference;High and medium concentration of berberine group and PBS group,cisplatin group,low concentration of berberine group A549 / DDP cells absorb light values have a statistically significant difference(P <0.01)after 48 h,72 h,,but with no statistical difference between high concentration and medium concentration of berberine group.(2)The high concentration of berberine in DDP group,low apoptosis rate of combined cisplatin group were higher than that of the PBS group,and there are statistically significant differences(P < 0.01),but the concentration of berberine group and comparison between the two groups,the high concentration of berberine group cell apoptosis rate was no statistical difference(P > 0.05).(3)Compared with the PBS control group,cisplatin group,low high berberine combined cisplatin group promoting apoptosis related proteins caspase3,caspase9,baxexpression obviously increased(P < 0.05),andcisplatin group,low high berberine combined cisplatin group between each other is also statistically significant(P < 0.05);Expression of BCL-2 antiapoptotic proteins decreased significantly(P < 0.05),and cisplatin group,low high berberine combined cisplatin group between each other is also statistically significant(P < 0.05).(4)Compared with the PBS control group,DDP group,low high concentrations of berberine in combination with cisplatin in promoting apoptosis related m RNA caspase3,caspase9,bax expression raised obviously,statistically significant(P <0.05),DDP group,low high concentrations of berberine in combination with cisplatin in antiapoptotic m RNA significantly lowered the BCL-2 expression,statistically significant(P < 0.05).Conclusion:Berberine in combination with cisplatin,the molecular mechanism of inhibiting proliferation and promoting apoptosis of A549 / DDP cells may be raised to up-regulate the expression of apoptosis related proteins caspase3 and caspase9,bax,lower resistance to apoptosis protein expression of BCL-2.