Study On Proteomics Of K562 Cells Treated With The Extracts From Patrinia Heterophylla

ObjectiveTo study the effects of the extracts from Patrinia heterophylla Bunge on Gene Expression Patterns during Morphogenesis of Chicken limb Buds in vivo. At the same times, to further explore the molecular mechanism of the extracts from Patrinia heterophylla on luckemia K562 cells by analyzing differences in global protein expression in K562 cells treated with or without the extracts from Patrinia heterophylla in vitro and mass spectrum identification.Methods1. Implant a bead into an chicken embryo, which was soaked in the extracts from Patrinia heterophylla Bunge. Detect the extracts-induced morphogenesis changes(Myf5, MyoD and PCNA).2. First, we cultured the human K562 cell in vitro. Make a control of the two groups:the experimental group was treated with the extracts from Patrinia heterophylla(100 ug/mL) and the control group was not treated with any drugs. The total proteins of control and Patrinia heterophylla-treated K562 cells were analyzed by two-dimentional gel electrophoresis (2-DE) after acetone purification and compared with those untreated K562 cells. After Coomassie staining and images were acquired by Image-scanner and then analyzed with the Image Maste software. We can established two-DE maps of control and Patrinia heterophylla-treated K562 cells.The differential proteins were identified by mass spectrometry. By MALDI-TOF-MS, we can analyse Partial differentially expressed protein spots and get peptide mass fingeiprinting and NCBInr database searching by Mascot software were used for protein identification.Results1. The extracts from Patrinia heterophylla Bunge (200mg/mL) can affect limb bud development, reduce gene expression of Myf5, MyoD and PCNA.2. We were obtained well-resolved,reproducible 2-DE patterns of control and Patrinia heterophylla-treated K562 cells. In the control group and experimental group, average spots of 3 gels were 378±16å'Œ397±21.Subtracting background,In the control group and experimental group, average spots were 328±15 and 352±12. Matching ratio were 86.77% and 88.66%. 3.2-DE images of the extracts from Patrinia heterophylla-treated and untreated K562 cells were analyzed.24 differential protein spots were found.11 differential protein spots were identied by masss pectrometry, including eukaryotic translation initiation factor 5A-1, Moesin,,α-tubulin, heat shock protein HSP 90, aging-associated gene 9 protein, hemoglobin, glyceraldehyde-3-phosphate dehydrogenase, aldolase A, flavin reductase.Conclusion1. Chicken limb development can serve as a convenient in vivo model for studying the effect of the extracts from traditional Chinese medicine on gene espression.The extracts from Patrinia heterophylla Bunge can inhibit cell differentiation and proliferation.2. Extracts from Patrinia heterophylla may inhibit the proliferative activtives of K562 cells and exert anti-K562 function by change the protein expression of K562 cells.