The Effect Of Piperine Combined With Cisplatin On Proliferation And Expression Of MMP-2 And VEGF In Lung Adenocarcinoma A549 Cells

Objective:To observe the effects on the growth and apoptosis of A549 cells as well as the expression of MMP-2 and VEGF in monotherapy group(Piperine or Cisplation alone) or combination group(Piperine combined with Cisplation) with a series of concentrations.We explored possible synergies and action mechanisms in order to provide a new clue and strategy to treat lung cancer in the application of combination group.Methods:1.MTT assay was used to detect the inhibition effect on A549 cells with different concentrations of piperine(12.5,25,50,100ug/ml), cisplatin(0.5,1,2,4ug/ml) and the combination group(50ug/ml piperine and 2ug/ml cisplatin) at 24 h, 48 h, 72 h.2.The Flow Cytometry was performed to assess the cellular apoptosis rate of A549 cells with piperine(50ug/ml), cisplatin(2ug/ml) and the combination group(50ug/ml piperine and 2ug/ml cisplatin) at 48 h.3.A549 cells were treated respectively with piperine(50ug/ml), cisplatin(2ug/ml),combination group(50ug / ml piperine and 2ug / ml cisplatin) during 48 h, then the expression of MMP-2 and VEGF m RNA was measured by RT-PCR.Results:1.Both Piperine(12.5,25,50,100 ug / ml) and cisplatin(0.5,1,2,4ug / ml) have inhibition effect on the growth of A549 cells,which showing that the inhibition effect was enhanced in a time-dependent and concentration-dependent pattern,while a significant difference in the control group(P <0.05).The synergistic antitumor effect of combination group(50ug / ml piperine and2 ug / ml cisplatin) is more obviously than piperine(50ug / ml) or cisplatin(2ug / ml) alone(P<0.05), of which the combined group presents extremely significant inhibition effect at 72 h.2.When Piperine(50ug / ml), cisplatin(2ug / ml) and the combination group were treated A549 cells after 48 h, the apoptosis rate was(10.67 ± 0.97)%,(16.56 ± 0.56)%,(55.52 ± 1. 33) %,which showing significantly higher than the control group(0.33 ± 0.55)%(P <0.05).Compared with piperine or cisplatin alone,the apoptosis rate of the combination group is significantly improved(P <0.05).3.Piperine(50ug / ml) and cisplatin(2ug / ml) were treated respectively A549 cells after48 h,the expression of MMP-2 m RNA and VEGF m RNA was decreasing when compared with the control group(P <0.05).The inhibition effect on MMP-2 m RNA and VEGF m RNA expression in combination group is more obvious than other groups(P <0.05).Conclusions:1.Both Piperine and cisplatin have inhibition effect on the growth of A549 cells,and there is a time-dependent and concentration-dependent relationship.we should note that the combined effect of two drugs was remarkable,when compared with the monotherapy group.2.Both Piperine and cisplatin can induce the effect of apoptosis on A549 cells.The apoptosis rate of combination group was significantly higher than monotherapy group.3.Both Piperine and cisplatin inhibit the expression of MMP-2 m RNA and VEGF m RNA in A549 cells.In addition,the inhibition effect on the combination group is the most obvious among them,and then may prevent lung cancer metastasis and angiogenesis.