| Objective:This study was designed to explore about apoptosis mechanism of arsenic trioxide in bone marrow cells from patients with non-M3 Acute myelogenous leukemia(AML) and apply with a experience for cure AML in clinical as a new method.Methods:1. morphological observation:To determine cell morphology by light microscope after treatment with arsenic trioxide in different concentration(0, 0.5,1.0,2.0umol/l)at indicated time(24h,48h,72h).2. Flow cytometric analysisThe apoptosis rate were detected by flow cytometry after the bone marrow cells were treated with arsenic trioxide in different concentration(0, 0.5, 1.0, 2.0umol/l)at the 72nd hour.3. Methlation of p16 gene in the bone marrow cells was detected by the nested-methylation specific polymers chain reaction and the expression of p16 gene mRNA was determined in the bone marrow cells by semi-quantity reverse transcription polymers chain reaction.Results:1. morphological observation showed characteristic apoptosis changes including cell shrinkage chromatin condensation DNA fragmentation,membrane blebbing and formation of apoptotic bodies.2. FCM showed : The effect of As2O3 on the apoptosis rate of the bone marrow cells was concentration-dependent. And at the concentration of 2.0μmol/l, the apoptosis rate could reach a peak.4. P16 gene fail to express in bone marrow cells from patients with AML after methylation, the expression was recovered after the cells exposed to As2O3. And the effect of As2O3 was concentration-dependent.Conclusions:1. As2O3 may activate the expression of p16 gene by demethylation in bone marrow cells from patients with Acute myelogenous leukemia.2. As2O3 could induce apoptosis of bone marrow cells from patients with AML and the effect of As2O3 was concentration-dependent.
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