Protein Profiling Of Hl-60 With High Tumorigenicity Developed Through Nude Mice Model

Objective: The tumorigenicity of Human tumor cells in nude mice was an important indicator of characteristics of tumor biology. And it had been widely used in various tumor pathogenesis and therapeutic research. We established the high tumorigenic HL-60 in nude mice and used differential proteomics approach to screen and identify differentially expressed proteins, hoping to give some thoughts for the molecular mechanism of the tumorigenicity of Leukemia cell lines .Methods: The high tumorigenic HL-60 in nude mice was established by combining implantation in vivo and culture in vitro. The fluorescence differential In-gel eletrophoresis(DIGE) was used to compare the differentially expressed proteins with the increasing of tumorigenicity. DeCyder6.5 was applied to select the significantly up-regulated or down-regulated protein spots. And the protein spots were identified by MALDI-TOF MS or MS/MS.The cellular localization and function of the identified proteins were analysis according to the comment Information of Swiss-Prot/UniProt.Results: 1.HL-60 model with high tumorigenicity in nude mice have been Successfully established. We used HL-60 to transplant firstly,and its tumorigenic rate was 30%. The second transplantation was conducted with HL60-1,and the tumorigenic rate was 50%. The third transplantation with HL60-2, and the tumorigenic rate was 80%. The last with HL60-3, and tumorigenic rate was 100%. We obtained HL60-4 with hign tumorigenic rate.2. Compare the differentially expressed proteins in common type HL-60 cell lines and high tumorigenic HL60-4 by DIGE, 87 protein spots with Significant difference were screened. 64 protein spots in which were identified as 24 gene product by MALDI-TOF-MS. 10 protein of them was up-regulated with the increasing of tumorigenicity, otherwise, 13 down-regulated. The differentially expressed proteins could been classified based on their functions:nucleic acid binding protein,cytoskeletal protein,chaperone,transcription factor,transferase,Protein metabolism and modification,carbohydrate metabolism,amino acid metabolism,and so on. The most important finding was:Heterogeneous nuclear ribonucleoprotein H appeared regular upward trend in HL-60,HL60-1, HL60-2, HL60-3, HL60-4cell lines, otherwise, Glutathione S-transferaseπand enolase 1 were regular downward trend.Conclusion: 1. In this study, the DIGE technology was applied to prepare protein expression of differential tumorigenic HL-60 cell lines, and we mastered the basic methods of proteomics research.2. The DIGE technology was used to screen, identify and analyse three tumorigenic-related differentially expressed proteins:hnRNP H,GSTπ,enolase 1。the upward or downward proteins play a facilitating or inhibitory in the occurrence and development of tumorigenicity of HL-60 cell line in nude mice. In-depth study of these proteins may provide the basis for understanding the occurrence and development of tumorigenicity of Leukemia cell lines .