Jieduhuayu Square Resistance Reversal Effect Of The Leukemia In K562/a02 Hl60/adr Cell Research

Objective:Research the regulative mechanism about the NF-kappa signal passage joining leukemia cells by using Molecular Bio-techniques, anti-tumor drug induction Nuclear factor-kappa B activation and Survivin expression, Suppresses the leukemia cell accent to perish, thus produces the process of multidrug resistance and Nuclear factor-kappa B activates mdrl to cause p-gp to express increases causes to the mechanism of multidrug resistance. Taking the signal passage of Nuclear factor-kappa B as the target spot, researched traditional Chinese medicine compound prescription Jiedu Huayu Chinese herbal serum to reverse leukemia multi-drug resistance target tropism.Methods:①Cell culture:K562, K562/A02, HL60, HL60/ADR cells vaccinates separately in contain 5% embryo cow blood serum's RPMI1640 nutrient fluid, under 37℃, the 5%CO2 condition raises, takes the logarithm vegetal period cells,2-3 days transfer generation one time, replace cell sap;②MTT examination Jiedu Huayu Chinese herbal serum dialog hemopathy K562, K562/A02, HL60, HL60/ADR cells multiplication influence; The Jiedu Huayu Chinese herbal serum reverse multi-drug resistance multiple determination;③Flow Cytometry measure cells to perish weakly and the DNR density of the leukemia multi-drug resistance cells K562/A02, HL60/ADR cells;④The Polymerase chain reaction (PCR) (RT-PCR) determinate the affect of Jiedu Huayu Chinese herbal serum to the expression multi-drug resistance gene Mdrl and the Survivin protein;⑤determine the research of Jiedu Huayu Chinese herbal serum to Nuclear factor-kappa B activeness by Using Western blotting (immunity signature method);With Western blotting technology examinate after Jiedu Huayu Chinese herbal serum processing multi-drug resistance cells NF-κB/P65, NF-κB/P50, IκBαof expression, and take the sensitive cell to as the comparison.Results:①The Jiedu Huayu Chinese herbal serum function after K562/A02, the HL60/ADR cell, in the cells the supple erythromycin's average fluorescence intensity has varying degree increase, therefore increases in the cells the supple erythromycin density is Jiedu Huayu Chinese herbal serum reverse K562/A02, the HL60/ADR cells including Jiedu Huayu Chinese herbal serum to resistance one of medicine mechanisms.②The Jiedu Huayu Chinese herbal serum to be possible to increase K562, K562/A02, HL60, HL60/ADR obviously cell's weak perishing rate, therefore the promotion target cell perishes is also weakly the Jiedu Huayu Chinese herbal serum to reverse K562/A02, the HL60/ADR cells to resist one of multidrug mechanisms.③Jiedu Huayu Chinese herbal serum after the RT-PCR determine Jiedu Huayu Chinese herbal serum to be able to decline K562/A02, HL60/ADR cells to perish weakly suppresses protein Survivin and the Mdrl multi-drug resistance gene of expression.④Jiedu Huayu Chinese herbal serum after the Western-blotting technology determine Jiedu Huayu Chinese herbal serum to be able to cause K562/A02, HL60/ADR cell NF-KB/P65, NF-KB/P50, the IKBa expression dropped.Conclusion:①The Jiedu Huayu Chinese herbal serum can reverse leukemia K562/A02, the HL60/ADR cell's multidrug resistance.②The Jiedu Huayu Chinese herbal serum reverse leukemia K562/A02, HL60/ADR cells to the mechanism of multidrug resistance may be related with the following:by enhancing the accumulated density of DNA in the leukemia K562/A02, HL60/ADR cells; promoting K562/A02, HL60/ADR cells to perish weakly; decreasing the expression of protein Survivin and the Mdrl (multi-drug resistance gene); Suppressing Nuclear factor-kappa B signal passage's activeness.