Differential Protein Analysis Of HL-60 Cell Xenograft Tumor In Nude Mice

As the blood system cancer, leukemia cells have the characteristics of cellular heterogeneity. The HL-60 cells, commonly used human myeloid leukemia cell line, have displayed the character of malignant phenotypes and shown the heterogeneity in cell morphology in cell cultures. What can be sure is, with the increase of emerging rate of tumor, various biological characteristics of high tumorigenicity HL-60 cell must change, but the internal mechanism is unknown, and also not been reported. As a powerful tool of differential expression protein comprehensive analysis, proteomics has been widely applied to diseases, especially cancer research. In view of this, we focused on high tumorigenicity HL-60 cells to study differential proteomics, to explore the mechanism of the increase of tumorigenicity rate. This can provide important value research data for the understanding of leukemia pathogenesis, progression and prognostic of the disease.In this study, two-dimensional electrophoresis DIGE technology was used to obtain differential expression protein between HL-60-G1 (tumorigenicity rate 30%), HL-60-G2 (tumorigenicity rate 50%), HL-60-G3 (tumorigenicity rate 80%), HL-60-G4 (tumorigenicity rate 100%) and ordinary HL-60 cell. DeCyder 6.5 image analysis software was used to select significant differential expression protein (ratio≥1.5,p≤0.01), and mass spectrometry was used to identify differential expression proteins. Protein cellular localization and functional analysis were analyzed under the Swiss-Prot database annotation information. Finally, Western blot and immunohistochemisty were used to verify some proteins.In this study, 87 differentially expressed spots were selected, in which 33 protein spots were significantly up-regulated in high tumorigenic cells, whereas 54 spots were down-regulated. Using Mass Spectrometry analysis, among the 32 significantly up-regulated spots in the high tumorigenic cell group, 25 spots corresponding to 11 different gene products were identified. Among the 55 significantly down-regulated spots in the high tumorigenic cells group, 40 spots corresponding to14 different gene products were identified. Protein cellular localization and functional analysis were analyzed under the Swiss-Prot database annotation information, and we found that the up-regulated protein mainly located in nuclear, whereas the down-regulated protein mainly located in cytoplasm. While the main function of these proteins are divided into: binding, catalytic, transcription regulation, transport and so on. Using Western blot technique and immunohistochemistry, respectively, two up-regulated protein T-complex protein submit alpha and Heterogeneous nuclear ribonucleoprotein H1, two down-regulated protein Enolase 1 and Glutathione S-transferase P were verified, the results are better with the DIGE results consistency.In this study, proteomics technology was successfully used to identify, analyze and verify a number of differential expression proteins, which associated with increased tumorigenicity. These up-regulated or down-regulated proteins play an important part in the occurrence and development of the increase of tumorigenicity. The further study of these proteins can provide evidence to the occurrence and development of acute myeloid leukemia.