The Proliferation And Apotosis-Inducing Effect Of Tongbu-Xiaoji Decoction To A549Cells

Objectives:To investigate the influence of the Tongbu-Xiaoji decoction to the human lung adenocarcinoma cell line A549cells, and the expression of PCNA and Survivin which are correlated with proliferation and apoptosis. To discuss the mechanism of the Tongbu-Xiaoji decoction to lung carcinoma.Methods:(1) The cell morphological effect of Tongbu-Xiaoji decoction to the human lung adenocarcinoma cell line A549cells was observed with inverted microscope.(2) The sensitivity and growth curve of the contained herb-serum to A549cells were observed with MTT.(3) The mRNA expression of PCNA and Survivin in cells were evaluated by RT-PCR, and the protein expression of PCNA and Survivin in cells were evaluated by Western blot.(4) The different categories of A549cell lines were stained with PI and Annexin V, and were evaluated by flowcytomety.Result:(1) There were some cell morphological change of herb serum containing Tongbu-Xiaoji decoction treating the A549cell line in24-48hours, cell volume became small, cell became irregular and apart of cells fall off. The cell numbers of A549reduced materially in72hours, and some cell began to confluence, the cell straining became deep, cell body began smaller.(2) The herb serum containing Tongbu-Xiaoji decoction showed growth inhibiting visibly to cell line A549. And there were dose-dependent and time-dependent manner in some time point,that is the proliferation was significantly enhanced with increasing reaction time and serum drug dose.The optimal affection time of herb serum containing Tongbu-Xiaoji decoction on cell was in72hours. The inhibition rate was38.63%.(3)The herb serum containing Tongbu-Xiaoji decoction showed apoptosis inducing ability to human lung adenocarcinoma in cell line A549. A549cells were treated by Tongbu-Xiaoji decoction with low、median and high dose medicated serum, Compared with that of blank group, there was a significant dlfference(P<0.01).Tongbu-Xiaoji decoction shows a significant apoptosis inducing effect to A549,and the affection shows a dose-dependent and time-dependent manner,that is the apoptosis inducing effect was significantly enhanced with increasing reaction time and serum drug dose.The optimal affection time of herb serum containing Tongbu-Xiaoji decoction on cell was in72hours. The apoptosis inducing rate was36.92%.(4) From the results of RT-PCR,A549cells were treated by Tongbu-Xiaoji decoction with48h, the relative values of PCNA mRNA expression and Survivin mRNA expression were reduced significantly(P<0.05),and the affection shows a dose-dependent manner. The protein expression of PCNA and Survivin are as the mRNA expression, Tongbu-Xiaoji decoction restrains the protein expression of PCNA and Survivin of A549cells significantly(P<0.05),and the affection shows a dose-dependent manner.Conclusions:(1) Tongbu-Xiaoji decoction significantly inhibited the proliferation of to A549cells, and the effect had an obvious dose-effect relationship in a certain period of time.(2) Tongbu-Xiaoji decoction restrains the mRNA and protein expression of PCNA to A549cells significantly.(3) Tongbu-Xiaoji decoction has the apoptosis-inducing effect to A549cells.(4) Tongbu-Xiaoji decoction restrains the mRNA and protein expression of Survivin to A549cells significantly.