| Objectives:The purpose of this study is to investigate the effects of Angelicasinensis(Oliv)Diels compound injection, Ephedrine, Vauqueline and Strychnine on human leukemic multidrug resistance(MDR) in K562/A02 cell line MDR1(multidrug resistance 1), MRP(multidrug resistance-associated protein), Topoâ…¡(topoisomeraseâ…¡), GST-Ï€(glutathione s-transferase) genes and their proteins.Subject:Human leukemic cell line: K562/A02.Methods:The Trypan blue assay was used to analyze the cytotoxic effect of Angelicasinensis (Oliv) Diels compound injection and the MTT assay was used to analyze cytotoxic effect of the other traditional Chinese medicine. The expression of MDR1, MRP, Topoâ…¡, GST-Ï€and their proteins in leukemia K562/A02 cells were respectively detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western Blot.Results:1. The groups respectively treated with Angelicasinensis(Oliv)Diels compound injection, Ephedrine and Vauqueline, Semi-quantitative RT-PCR showed that the mRNA of MDR1 gene were all reduced significantly (P<0.01); and Western Blot also showed that the expression of P-gp were decreased (P<0.01). The mRNA of MDR1 and the expression of P-gp in leukemic K562/A02 cells treated with Strychnine were not conspicuously changed (P>0.05).2. The group treated with Angelicasinensis(Oliv)Diels compound injection, Semi-quantitative RT-PCR showed that the mRNA of MRP, Topoâ…¡, GST-Ï€genes were not significantly changed (P>0.05); Western Blot show that the expression of topoisomeraseâ…¡protein was increased (P<0.01); the other proteins were not significantly changed (P>0.05). The group treated with Strychnine, Semi-quantitative RT-PCR showed that the mRNA of MRP gene was reduced significantly (P<0.01), the other genes were not significantly changed (P>0.05); and Western Blot showed that the expression of MRP protein was decreased (P<0.01); the other proteins were not conspicuously changed (P>0.05). The groups respectively treated with Ephedrine and Vauqueline, the mRNA of MRP, Topoâ…¡, GST-Ï€and their proteins were not significantly changed (P>0.05).Conclusions:1. Angelicasinensis(Oliv)Diels compound injection, Ephedrine, Vauqueline could partly reverse the multidrug resistance in K562/A02 cells. Decrease of MDR1 gene and P-glycoprotein expression maybe is one of the MDR reversal pathways in K562/A02 cells.2. Angelicasinensis(Oliv)Diels compound injection could increase expression of topoisomeraseâ…¡protein, while the mRNA of Topoâ…¡gene was not affected. And the MRP, GST-Ï€genes and their proteins were not affected by treatment of Angelicasinensis(Oliv)Diels compound injection. Ephedrine and Vauqueline were not affected the expression of MRP, Topoâ…¡, GST-Ï€genes and their proteins. Strychnine may suppress the expression of MRP gene and its protein, while Topoâ…¡, GST-Ï€genes and its proteins were not affected after treatment of Strychnine. |
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