Study For Artesunate On The Inhibition And Mechanisms Of Human Acute Leukemia Primary Cells Procedures

Background:Acute leukemia is a malignant tumor in hematopoietic system. The mortality rate is high. In recent years, with the further study, the chemotherapy, bone marrow transplant and molecular biology treatment method for the leukaemia had improved remission rate obviously, even be cured. But the strong poisonous effect, multidrug resistance and the expensive cost, limited marrow providing bodys, and transplant bring serious neurological complications limited therapy of modern medicine.Since the 1980s, some researchers of TCM used traditional chinese medicine extracts to treat acute leukemia. Had gain a noticeable effection. Art as an plants can anti-malarial in clinical application. In 2007, under the guidance of professor Yang Hongyong, we have used Art to experimentalize on K562, HL-60 cell line and 16 cases of acute leukemia patients.The results all showed that:Art can inhibit the growth of human acute leukemia primary cells.But its the mechanism and the target are up for exploring further.Objective:This study is on the basis of research results in the past, by establishing the training system with human acute leukemia primary cells, in contrast with DUR and the saline water in the blank, Observe the effects of different concentration of Art on the proliferation of the primary cells, Provide the preliminary support for Art to treat leukemia from cell cycle,apoptosis and oncosis,molecular biology gene and cell genetics.Methods:11 patients who were all diagnosed as acute leukemia were recruited into research. Establishing the training system in vitro with their bone marrow primary cells. Cells were cultured in logarithmic growth phase were seeded in 96,12 and 6 well plates Respectively.To a final concentration of 50 u g/ml,25μg/ml,12.5μg/ml of Art, concentration of 25μg/ml DNR and the same dosage of saline solution for a certain time. Using the method of MTT to detect the effect of the proliferation cells; Using electron microscope to observe the forms of apoptosis and onconsis; Using Western Blotting testive technology to detect the protein of the cell apoptosis; Using Gene chip testive technology to detect the perform expression of AL related genes. To explore the mechanism and target for Art on human acute leukemia.1. MTT detective results showed that:Art can inhibit the growth of human acute leukemia primary cells. And the inhibition ratio is dependent on the dose of Art and the functionary time.2. Electron microscopic observational results showed that:Art can cause the different degrees of the apoptosis and and oncosis of leukemia primary cells after 48 hours. And the degree of apoptosis and and oncosis is dependent on the dose of Art.3. Western Blotting testive results showed that:Art can down-regulate protein Bcl-2;at the same time up-regulate protein Bax and cyc-c.4. Gene chip testive results showed that:Art can perform expression of AL related genes.Conclusion:Art can inhibit the growth of human acute leukemia primary cells definitivly. Mitochondrial channe may be the main way of Antileukemic function and reversion of multidrug resistance. So Art is worth to be developed to the new drug of Antileukemia.