Effects Of Dobutamine On Biological Behavior And The Expression Of Yap Protein Of Human Lung Adenocarcinoma A549 Cells Treated With Cis-diamminedichloroplatinum

Objective:To investigate the effect of dobutamine on biological behavior of human lung adenocarcinoma A549 cells treated with cis-diamminedichloroplatinum(DDP).Methods:Human adenocarcinoma A549 cell line was cultured to experiment.The cells were randomly divided into control group,dobutamine group,DDP group and dobutamine +DDP group.The experiments were as follows:(1)The inhibitory rate of proliferation of the groups interfered with dobutamine(0,7.5,15,30,60,120?mol/L)or DDP(0,1.25,2.5,5,10,20?g/ml)24,48 and 72 h was detected by CCK-8 assay.And we selected appropriate concentration and action period to complete the experiment.(2)The scratch healing experiment was used to test the migration and movement capacity of the groups.(3)The invasion abilities of the groups were decided by transwell methods.(4)The apoptosis rate of the groups was analyzed with flow cytometry.(5)The protein expression of Yes-associated protein(YAP)and phosphorylation of YAP(phospho-YAP,P-YAP)in the groups was determined by Western Blot.Results:(1)CCK-8 cell proliferation assay: dobutamine or DDP could inhibit the proliferationof A549 cells in a concentration-and time-dependent manner(P<0.05).(2)The migration and movement capacity assay: compared with control group,the scratch healing rate of dobutamine group was reduced(P<0.05).Compared with dobutamine or DDP alone group,the combination of dobutamine and DDP significantly reduced the scratch healing rate(P<0.05).(3)The in vitro invasion abilities assay: compared with control group,the number of cells penetrating the matrigel of dobutamine group was reduced(P<0.05).Compared with dobutamine or DDP alone group,the combination of dobutamine and DDP significantly reduced the number of cells penetrating the matrigel(P<0.05).(4)Apoptosis assay: compared with control group,the cells apoptosis rate of dobutamine group was reduced(P<0.05).Compared with dobutamine or DDP alone group,the combination of dobutamine and DDP significantly reduced the apoptosis rate(P<0.05).(5)The protein expression of YAP and P-YAP assay: compared with control group,the protein expression of YAP of dobutamine and DDP alone group was decreased and the protein expression of P-YAP of dobutamine and DDP alone group was increased(P<0.05).Compared with dobutamine group,the combination of dobutamine and DDP significantly decreased the protein expression of YAP and increased the protein expression of P-YAP(P<0.05).Conclusion:Dobutamine enhances the DDP effect through descending the expression of YAP to restrain the proliferation,invasion,migration and to enhance the apoptosis of A549 cells with a synergistic method.