| Objective: Transfusion has become an important therapy in clinical practice. The transfusion requirements of safety, scientific, reasonable and effective requires us to have more clear understanding of the composition of the blood products. Currently,fresh frozen plasma and cryoprecipitate are often transfused to patients.Plasma may contain different levels of microparticles due to individual differences in blood donors.This study was to investigate the effect of fresh frozen plasma from different blood donors on tumor cell proliferation in vitro.Mehtods:After HepG2 and AGS cells were c?ltured with the fresh frozen plasma from different male blood donors in blood transfusion department of Chinese PLA General Hospital for 24 hours. The methyl thiazolil tetracolium (MTT) and real-time cell-based assay(RTCA) were used to measure the proliferation of tumor cell in vitro and screened the different FFP which promoted the proliferation of tumor cells significantly different. In this study Luminex method was applied to test the levels of TNF-a, IL-lb, IL-6, VEGF, MCP-1, RANTES and PDGF-AB/BB in fresh frozen plasma. Application of SELDI/ TOF-MS technology study related protein expression in fresh frozen plasma to find the key proteins.Res?lts:(1) The tumor cells were treated with different concentrations of mixed FFP from different blood donors for 24h. In vitro study of HepG2 cells proliferation obviously after treated with mixed FFP in dose-dependent manners. Compared to control group,there were no significant differences between different age groups when final concentration was 10%. The experiment of HepG2 cells proliferation indicated that the mixed FFP of the cased with ages of over 50 years old produced more proliferation of tumor cells with OD value 0.629,0.699 and 0.821(final concentration were 20%,30% and 40%) than that of 21?30 years old group with OD value 0.453,0.538 and 0.584(P=0.0298?P=0.0203 and P=0.0241). The MTT method was used to measure the effect of FFP from different blood donors on proliferation of HepG2 cells in vitro. The OD value 0.964 of tumor cells from no 14 FFP group were higher than that of no 23 group with OD value 0.677(P=0.0000). Different FFP from different blood donors promoted the proliferation of HepG2 tumor cells in vitro significantly different.(2) The MTT and RTCA methods were used to measure the proliferation of HepG2 tumor cells after treated with FFP form different blood donors in vitro. Through statistical analysis processing, the res?lts of two kinds of methods were no statistically significant difference. And RTCA method was used to measure the proliferation of HepG2 and AGS tumor cells after treated with FFP form 68 different blood donors in vitro. The experiment of cells proliferation indicated that the FFP of the 68 cased with HepG2 tumor cells was consisted with AGS tumor cells.(3) The res?lt of RTCA assay showed that the tumor cells proliferated differently after treated with FFP from different blood donors. And screened the different FFP which promoted the proliferation of tumor cells significantly different and with high cell index(CI). Luminex method was applied to test Luminex method was applied to test the levels of TNF-a, IL-lb, IL-6, VEGF, MCP-1, RANTES and PDGF-AB/BB in fresh frozen plasma. The level of PDGF-AB/BB in FFP with CI level higher than 7.0 signifcantly higher than that of FFP with CI level lower than 3.0. Application of SELDI/TOF-MS technology study related protein expression in fresh frozen plasma with CI higher than 8.0 and lower than 3.0 to find the key proteins. There were 26 proteins expressed differently, including Metastasis-suppressor gene 1, cortistatin,platelet factor 4, phosphatidylethanolamine-binding protein 1, matrix metallo proteinase-9, Lactotransferrin, transforming growth factor-a. With the increasing expression level of growth factors and proteins, the levels of growth factors increased in FFP. It may be factors proliferated HepG2 tumor cells in vitro.(4) The MTT method was used to measure the effect of mixed FFP with CI level higher than 7.0 and mixed FFP with CI level lower than 3.0 on proliferation of HepG2 cells in vitro. Two groups of FFP were statistically significant differently.Application of RayBiotech atibody array technology study related protein expression in fresh frozen plasma.Smad 4,Glut2,VEGF,MIP 2,MMP-7,TIMP-l,ALCAM,RANTES,IL-lb,IL-6,MCP-1 and TNF-.a were remarkably upreg?lated in the proliferation group and 15 signal pathways had significant difference.Conclusion:(1) Different FFP from different blood donors promoted the proliferation of HepG2 tumor cells in vitro significantly different.(2) The different levels of growth factors increased in FFP may be factors proliferated HepG2 tumor cells in vitro. |
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