The Study On The Inhibitory Effect Of Docetaxel Combined Delisheng On The Proliferation Of Human Lung Cancer Cell A549 Cells And Its Mechanism In Vitro

Objective:To investigate the inhibition effect of Docetaxel, Delisheng and the combination of the two drugs on the proliferation,apoptosis and gene expression of non-small cell lung cancer A549 cell. To analysis the interactions form of the two drugs.Methods:MTT assay was used to determine the proliferation inhibition by Delisheng and Docetaxel to A549 cell. Annexin-V kit was used to test the early apoptosis of A549 cells induced by Delisheng and Docetaxel. The late apoptotic cells were assessed by propidium iodide (PI) staining. Semi-quantitative RT-PCR was employed to measure the expression of survinvin,VEGF,bFGF ,α5 intergrin submit ,β1 intergrin submit ,bcl-2 and bax in A549 cells .Results:(1)The survival rate of A549 cells in the Delisheng,Docetaxel,and the combination groups ,were significantly lower than that in the contol group(P<0.05), and the survival rate of A549 cells in the combination groups was lower than that in the Delisheng or Docetaxel group(P<0.05). The interaction between Delisheng and Docetaxel was synergy or additivity. (2)After treatment with Delisheng, Docetaxel and the combination of these two drugs for 24h, we could see the typical morphological changes of apoptosis of A549 cells. Further test results detected by flow cytometry show that the late apoptosis rate of A549 cell in the Delisheng,Docetaxel,and the combination groups ,were significantly higher than that in the contol group(P<0.05), and the late apoptosis rateof A549 cell in the combination group was significantly higher than that in any kind of single-drug (P <0.05).(3) RT-PCR results showed that the related expression level of survivin,β1 intergrin submit mRNA were lower in the cells treated with delisheng than that in untreated cells(P﹤0.05),while the related expression levels ofα5 integrin submit, VEGF and bFGF mRNA were not significantly decreased with delisheng treatment (P﹥0.05). (4) The related expression level ofα5 integrin submit mRNA and the ratio of gene expression bcl2/bax were lower in the cells treated with docetaxel than that in untreated cells(P﹤0.05),while the related expression levels of VEGF, bFGF,β1 integrin subunit mRNA were not significantly decreased with docetaxel treatment (P﹥0.05). (5) The related expression level of survivin,β1 intergrin submit mRNA in the A549 cells treated with delisheng combined docetaxel were not higher than that in A549 cells treated only with delisheng (P﹤0.05),and the related expression level ofα5 integrin submit mRNA and the ratio of gene expression bcl2/bax in the A549 cells treated with delisheng combined docetaxel were not higher in the cells treated with docetaxel only(P﹤0.05),while the related expression levels of VEGF and bFGF mRNA were not significantly decreased with delisheng combined docetaxel treatment (P﹥0.05)。that is, the interaction in the gene level between the two drugs as follows: two single-drug can play their own role without interfering, and their role are complementary.Conclusions:Compared with the Docetaxel or Delisheng,Docetaxel combined Delisheng can better inhibit the proliferation of non-small cell lung cancer A549 cells,and induce these cells to apoptosis. The suppressed expression of bcl-2/bax ,survivin,α5 intergrin,β1 intergrin may be the mechanisms of apoptosis induction.