The Experimental Studies On The Reverse Of Resistance Using Chansu In Multidrug Resistant Leukemic Cell

Objective:Chronic myeloid leukemia is the Primary malignant disease of Hematopoietic system,35 years of age the incidence of the highest mortality rate of malignant. K562 cells in chronic myeloid leukemia cell lines in a CML cell line,human hematopoietic stem cells to erythroid differentiation by Friends virus infection,to stop the formation of differentiation.After years of research, traditional Chinese medicine has a strong clinical advantage on the Treatment of leukemia.A lot of Chinese medicine treatment of leukemia has obvious effects.To study the effect of chansu on growing,the reverse and the apoptosis of the K562/ADR cells,in order to provide useful based of cytology,molecular biology for multidrug resistant leukemia.Methods:The main experiments is in vitro cell culture in this study,the study includes:(1) Chansu experimental serum pharmacological methodSerum pharmacology of Chinese medicine is a better way to study Chinese medicine,however,must distinguish between the different drugs and different levels of preparation program.At the same time in the study of pharmacological, The Chinese medicine can be combined with the Chemical serum,the active ingredients can be qualitative or quantitative analysis.This is the development of Chinese medicine is more practical significance.(2) Analyse the growth inhibition of the cells by MTT assay;From logarithmic phase cells,Adjust the cell number is 1×105/ml, Adding 100ul to 96' ELISA plate,Each set of parallel 8-hole,ELISA plate will be placed in 37℃,5%CO2,adding different concentrations of experimental drugs,training the different times,detection by the Automatic ELISA' OD. Calculated in accordance with the experimental group of drugs on the leukemia cell cytotoxicity.(3) Detect the cell apoptosis by flow cytometry;Collecting more than the role of the above of division,With Annex in V and PI to treat a series of cells,send their to FCM apoptosis rate in each group.(4) The molecular assay of RT-PCR was used to examine the mRNA level of gene Survivin mRNA.Extract the above division of cells' RNA.Reverse transcription reaction, Followed by RT-PCR reaction,to get the data.Through these observations,in order to reflect the molecular mechanism of the reverse of resistance using chansu in multidrug resistant leukemia.Results:(1)Treatment of K562/ADR cells with chansu inhibited the cell growth in a dose-dependent manner.(2)Apoptotic K562/ADR Cells were found after incubation with drugs.(3)Positive findings of Survivin mRNA decreased after incubation with drugs.(4)The percent of apoptotic K562/ADR cells increased after incubation with drugs.Conclusion:Chansu can inhibit the K562/ADR cell growth,reverse the resistance and induce the apoptosis in a dose-and time-depentent manner,while positive findings of Survivin mRNA decreased.