Study On Gene Expression Profilef Of K562 Cell After Treatment With Ginseng Polysaccharide Injection

Leukemia as hematopoiesis system of malignant tumor, clinical diagnosis and treatment, mainly due to the difficulty is often a lack of particularity; etiology The existing subtypes, molecular level of clinical classification difficulty, can't targeted medication; Traditional chemotherapy for the larger, usually can ease damage condition, but cannot improve hematopoiesis function and cure, eventually patients die of hematopoietic and immune function failure. Recently, research of differentiation of leukemia, especially from traditional Chinese medicine screening effective differentiation agent is hotspot.Panax Ginseng is the "traditional Chinese medical tonifying qi" to medicine. Ginseng Polysaccharide Injection, the abbreviation Ginseng Polysaccharide. From ginseng extract of glucose, such compounds in polymer tumor in treatment and prevention has good activity, but the specific molecular mechanism has mostly clarified. Therefore, this subject adopts experiment hematology techniques, the GPSI on red leukemia cell (K562) inhibitory effect, on this basis through gene microarray assay screening GPSI function of gene expression. Through the research of interpretation K562 cell proliferation of GPSI inhibiting, differentiation, promote the influence of apoptosis with gene expression, clarifies the correlation between changes in the role of ginseng polysaccharide injection molecular mechanism of leukemic cells for research and development, the natural medicine clinical applications of differentiation agent with theoretical basis and experimental basis.MethodsUsing four methyl azo thiazole salt (determined by MTT) method, GPSI for in vitro cell proliferation K562 role influences; Through the cell hemoglobin detection, Wright 's dyeing, LianBenAn dyeing, to study the GPSI K562 after induction to red granulorytes situation; Through the flow cytometric profilometry (FCM), detection by GPSI after induction; K562 apoptosis Gene chip technology screening ginseng polysaccharide K562 cell injection injection effect after gene expression differences, then the webgestalt GoTree classification, using biological function analysis.The results1. Ginseng polysaccharide injection of injection (GPSI) K562 cell proliferation has inhibitory role:GPSI in vitro K562 cell proliferation of an inhibitory effect, in 25-800 mg/L concentration range, its inhibition is concentration dependence, and they are all in action when 48h inhibition rate reaches peak. According to the measured MTT method, to determine the value analysis OD 400mg/L role for optimal 48h drug concentration.2. GPSI can induce K562 cell to mature direction differentiation:(1)The morphology observation shows that:Wright 'dyeing: K562 cell volume control is negatie larger, nuclear big and clear, nuclear qualitative proportion is large, strong basophilic cytoplasm, no special particles. The positive control Hm (30 umol/L), visible cell size decreased, nuclear decreased diameter, cytoplasm, nuclear plasma rate by rich. GPSI (400 mg/L) after induction, nuclear decreased diameter, cytoplasm, nuclear plasma rate by rich.Benzidine staining: Benzidine after dyeing visible: negative control group did not see K562 cell; the coloring The positive control Hm (30 u mol/L) and GPSI (400 mg/L) group of cells are visible blue hyperchromatic cells.(2) The hemoglobin determination: ginseng polysaccharide injection (GPSI) injection of K562 cell have in vitro induced hemoglobin generated role in 100-800mg/L concentration range, show concentration dependence, and they are all in action when 48h inhibition rate reaches peak.3. K562 cell apoptosis induced GPSI can AnnexinV/intraoperative: streaming cells dyeing method detection, double PI control group and GPSI 200 mg/L, GPSI 400 mg/L respectively K562 cell 48h role, apoptosis rate, respectively 7.095 3.465 + 0.417, 0.898, 9.895 + 0.502 hospital, and the results showed that GPSI K562 cell, role of cell apoptosis increasing proportion occurred (P < 0.05).4. Detection, gene expression differences were found 306 differentially expressed genes, including 220 and down-regulate 86 express decline.5. GoTree genetic variations webgestalt analysis, classification after the 306 genetic differences were searching, and 267 a at the molecular function, biological processes, cell component of the function of form remarkable rich group.