Study On The Effect And Mechanism Of The Compatibility Of The Gentianella Acuta And The Hypericum Attenuatum Choisy On Human Lung Cancer A549 Cells

Objective:To observe the effect of the combination of Gentianella acuta and Hypericum attenuatum choisy on human lung cancer cell line A549 and the underlying mechanism.Method:We prepared drug-containing serum and cultured human lung cancer A549 cells in vitro.The experiment was divided into groups as follows:blank group,gentianella acuta group,hypericum attenuatum choisy group,gentianella acuta and hypericum attenuatum choisy 2:1 compatibility group,gentianella acuta and hypericum attenuatum choisy 1:2 compatibility group,gentianella acuta and hypericum attenuatum choisy 1:1 compatibility group and positive drug group(5-FU).MTT assay was used to detect the effect of drug-containing serum on the proliferation of human lung cancer A549 cells in each group.The effect of drug-containing serum in each group on the apoptosis rate of human lung cancer A549 cells was detected by flow cytometry.RT-PCR was used to detect the expression of Fas mRNA,Fasl mRNA,Caspase一8 mRNA,Caspase-3 mRNA,Bax mRNA,Bcl-2 mRNA,Cyt-c mRNA and Caspase-9 mRNA in human lung cancer A549 cells.The protein expressions of Fas,Fasl,Active Caspase-8,Active Caspase-3,Bax,Bcl-2,Cyt-c and Active Caspase-9 in human lung cancer A549 cells were detected by Western Blot.Result:l.MTT assay showed that compared with the blank group,the drug-containing serum in each group could significantly inhibit the proliferation of human lung cancer A549 cells and had a time-effect relationship.Compared with the gentianella acuta group and the hypericum attenuatum choisy group,the 2:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)had the highest inhibition ratio in the proliferation of human lung cancer A549 cells.2.The results of flow cytometry showed that compared with the blank group,the drug-containing serum in each group could induce the apoptosis of human lung cancer A549 cells.Compared with the gentianella acuta group and the hypericum attenuatum choisy group,the 2:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)had the highest apoptosis ratio in the human lung cancer A549 cells.3 RT-PCR results showed that the drug-containing serum in each group could significantly increased the expression of Fas mRNA,Fasl mRNA,Caspase-8 mRNA,Caspase-3 mRNA,Bax mRNA,Cyt-c mRNA and Caspase-9 mRNA and down-regulated Bcl-2 mRNA expression compared with the blank group.Compared with the gentianella acuta group,the 2:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)could significantly up-regulate Fas mRNA.Fasl mRNA.Caspase-8mRNA,Caspase-3mRNA,Bax mRNA,Cyt-c mRNA and Caspase-9mRNA and down-regulate Bcl-2 mRNA.Compared with the hypericum attenuatum choisy group,the 2:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)could significantly up-regulate Fas mRNA.Fasl mRNA、Caspase-8mRNA,Caspase-3mRNA,Bax mRNA,Cyt-c mRNA and Caspase-9mRNA and down-regulate Bcl-2 mRNA.The 1:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)could obviously up-regulate the Caspase-9mRNA and up-regulate Caspase-3 mRNA.4 Western blot analysis showed that the drug-containing serum in each group could significantly increased the protein expression of Fas,Fasl,Caspase-8,Caspase-3,Bax,Cyt-c and Caspase-9 and down-regulated Bcl-2 protein expression compared with the blank group.Compared with the gentianella acuta group,the 2:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)could significantly up-regulate the protein of Fas、Fas1、Caspase-8,Caspase-3,Bax,Cyt-c and Caspase-9 and down-regulate Bcl-2 protein.Compared with the hypericum attenuatum choisy group,the 2:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)could significantly up-regulate the protein of Fas、Fasl、Caspase-8,Caspase-3,Bax,Cyt-c and Caspase-9 and down-regulate Bcl-2 protein.The 1:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)could up-regulate the protein of Caspase-9 and Caspase-3.Conclusion:1 The drug-containing serum could inhibit the proliferation of human lung cancer A549 cells and induce the apoptosis.The effect of the the 2:1 compatibility group(gentianella acuta:hypericum attenuatum choisy)was the strongest.2 The mechanism of apoptosis induced by drug-containing serum in human lung cancer A549 cells is related to death receptor and mitochondrial signal transduction pathway.