| Objective:Lymphoma is one of the most common malignant tumors in blood system. Its pathogenesis is not clear, its diagnosis depends on tissue biopsy. But small lesions of early lymphoma orlocated in the deep lesions in vivo have lower detection rate of tissue biopsy. It lead to difficulties for clinical diagnosis. At present, the basic treatment strategies based on chemotherapy, and radiotherapy combined with comprehensive treatment. We use SELDI-TOF-MS technology for in vitro T, B lymphoma cell lines and normal lymphocytes to detect protein, We collect data through Proteinchip software, and use SPSS 13.0 statistical package for data analysis. Our purpose is look for the differences between normal cells and lymphoma cells and different protein differences between cell lymphoma protein. Although Cell lines in vitro may not fully reflect the in vivo cell growth status and the biological activity, it has the advantages of single component, good homogeneity, easy control of experimental conditions, etc. Through experiments we want to search for molecular markers of lymphoma.Furthermore study the expectations of the pathogenesis of lymphoma and for treatment, the initial target to lay the foundation, and help its early diagnosis. Methods: Cultivated conventional T, B lymphoma cell line in 25ml culture bottles of RPMI-1640 medium which contained 10% fetal bovine serum.Placed in a incubator with a humidified atmosphere of 5% CO2, at 37℃to cultivate, passaged them every 3-5 days. Used logarithmic phase cells for experiments. We used 6-hole plate inoculate two types of cells to continue to foster. and we collect cells respectively after 2,4,6 days of inoculation.There were 3 duplicate well in each time period. We used lymphocyte separation medium separate the normal lymphocytes from peripheral blood and inoculate it in the RPMI-1640 medium which contained 10% fetal bovine serum.Added PHA of 100ug/ml in it. We collect cells respectively after2,4,6 days of inoculation.There were 3 duplicate well in each time period in the same. We used protein extract extract from the total protein of various cells in each hole, Make respectively a little lymphoma cell culture medium on the mass spectrometry detection before collect cells. And applied surface-enhanced laser desorption - ionization -time of flight- mass spectrometry technology for detection, So we used CM10 chip with fingerprint access to protein expression. And we collected data through Proteinchip software, repeated the experiment three times,so we used SPSS 13.0 statistical package to preliminary statistical analyse data(calculate average?X, standard deviation S and P values)P <0.05 was considered statistically significant difference. . Results: 1. There are six differentially expressed protein peaks in T lymphoma cell lines and normal lymphocytes(P<0.05), there are five protein peaks in the T lymphoma cell line in high expression, and one Protein peaks in low expression. 2. There are six differentially expressed protein peaks in B lymphoma cell lines and normal lymphocytes(P<0.05), There are four Protein peaks in the B lymphoma cell line in high expression, And two Protein peaks in low expression. 3. B lymphoma cell lines and T lymphoma cell line has a common protein peak high expression and low expression than normal lymphocytes. 4. There are no significant differences in protein expression in T lymphoma cell lines of different culture time (in the short incubation time)5. There are no significant differences in protein expression in B lymphoma cell lines of different culture time (in the short incubation time). 6. There are seven differentially expressed protein peaks in B lymphoma cell lines and T lymphoma cell lines(P<0.05), Compared with B lymphoma cell line, There are three protein peaks in T lymphoma cell line in high-expression, and four peaks Proteins in low-expression.Conclusions: 1. The proteomics of T, B lymphoma cell line and normal lymphocyte have significantly different levels.2. The proteomics of T, B lymphoma cell line have significantly different levels.3. There are no significant differences in the level of proteomics in the same types of lymphoma cell line of different culture time(in the short incubation time).4. Application of SELDI-TOF-MS technology probably has significant value in screening molecular markers of lymphoma,moreover it can possibly lay the preliminary foundation in pathogenesis of lymphoma and for the initial target to treatment, , and help its early diagnosis. |
