Acute Lymphoid Leukemia Cell Lines Induced The Differences Of Protein Expression Using Methotrexate Enantiomers In Vitro

Objectives: To establish emthotrexate(MTX) enantiomers resistant human acute lymphoid leukemia CCRF-CEM and BALL-1 cell lines and analyze the acute lymphoid leukemia cell lines induced the differences of protein expression of MTX enantiomers in vitro.Methods: Using the method of low-dose induction combined with increasing concentrations to build MTX enatiomers resistant cell lines. To observe CCRF-CEM and BALL-1 cells in inverted microscope and the cells activity differences by Fluorescence microscopy. Using two-dimensional gel electrophoresis to separate the total protein of MTX enantiomers resistant cell lines and their parent cell lines, scanner was used to find out the different protein expression with a gel point and combined the mass spectrum analyzer to identify differences in protein expression with bioinformatics analysis.Results: The results of protein quantification were CCRF-CEM 36.17mg/ml, D-(-)MTX-CCRF-CEM 36.45mg/ml, L-(+)-MTX-CCRF-CEM 38.54mg/ml, BALL-1 41.09mg/ml, D-(-)MTX-BALL-1 37.28mg/ml, L-(+)-MTX-BALL-1 39.87mg/ml. The average protein spots separated from the parents cell-groups(CCRF-CEM and BALL-1) were 295±14 and 421±12 and the match rates were 82.57% and 85.13%. The clear protein spots separated from the treatment groups(D-(-)MTX-CCRF-CEM, L-(+)-MTX-CCRF-CEM, D-(-)MTX-BALL-1, L-(+)-MTX-BALL-1) were 316±10, 302±9, 445±11, 432±7 and the relative match rates were 85.27%, 84.72%, 86.89%, 89.35%. The volume of protein point difference ratio was more than two times the result was statistically significant. Compared with CCRF-CEM, D-(-)MTX-CCRF-CEM has15 significant different protein points, but L-(+)-MTX-CCRF-CEM only has 6. In the L-(+)-MTX-CCRF-CEM 6 different protein points, 4 were expressed in the treatment group and 2 were in the control group. D-(-)-MTX-BALL-1 has 17 different protein points compared to the BALL-1. L-(+)-MTX-CCRF-CEM has 12 significant different protein points, 10 were high expressed in the test group and the other two were expressed in the control group(BALL-1).Conclusions: The treatment groups(D-(-)-MTX and L-(+)-MTX) were expressed more protein points than the control groups(CCRF-CEM and BALL-1). The culture cells which resistant the D-(-)-MTX were expressed more protein points than the L-(+)-MTX. The cells of which resistant L-(+)-MTX were expressed less some normal cells protein.