Exploration Of The Effects Of STI571 On Mononuclear Cells And On Gene Expression Profile Of Acute Non-lymphocytic Leukemia In Vitro

The first chaptenThe influence of STI571, a selective tyrosine kinaseinhibitor, on the proliferation, differentiation, immunophenotypic and c-kitmRNA in the bone marrow cells from patients with acute non-lymphocyticleukemia in vitro.Objective: To explore the influence of STI571 on mononuclear cells fromthe bone marrow of ANLL patients with CD 117 overexprss in vitro,including proliferation, differentiation, immunophenotypic, c-kit mRNA andCD117.Materials and Methods: The samples are taken from 31 relapsed and initialANLL patients who were diagnosed. Mononuclear cells(MNC) were isolatedfrom these samples by Ficoll. A final concentration of STI571 is 0.1 n M,l uM,10n M which added once at the beginning of the cultivation period, andcontrol was set. Co-cultivate ANLL cells and STI571 for 72h in vitro, andobserved the effects on ANLL cells, including proliferation, differentiation.The expression of c-kit mRNA and GD117 were assayed by RT-PCR andflow cytometray respectively.Results and Disscussions:l.When treated the mononuclear cells with different concentration ofSTI571(0.1 u M,l u M,10 u M), the restraint of proliferation increasedgradually with significant statistics difference. And the longer treating time is,the more significant the restraint effect becomes.2. STI571 has the obvious effect of inducing cells toward iso-series maturedi- fferentiation in a dose-dependent manner.3.When treated the mononuclear cells with different concentration ofSTI571(0.1 U M,l P M,10 u M), the restraint of CD117 and c-kit mRNAchanged in a dose-dependent manner. Statistics difference appears indifferent concentrations and control groups and also in 0.114 M and 10 u Mgroup. The restraint effect shows its time-dependence during the observingperiod.The results of this experiment suggests in the concentrations ranging from 0.1 u M to 10uM, STI571 significantly restrains the proliferation , induces differentiation of ANLL with CD 117 overexpress and inhibits the expression of GDI 17 and c-kit mRNA in the cells from the bone marrow of patients with ANLL. It is concluded that STI571 has the obvious effect on the restraint of c-kit. It will provide the experiment guide for STI571 in treating the refractory or relapsed ANLL with GDI 17 overexpress. The second chapter: The study of gene expression profile in mononuclear cells from the bone marrow of patients with acute non-lymphocytic leukemia treated by STI571.Objective: To study the gene difference expression in mononuclear cells from the bone marrow of ANLL patients with GDI 17 overexpress treated by STI571,so as to explore initially the mechanism of the restraint of proliferation and inducing differentiation caused by STI571. Materials and Methods: The samples are taken from 2 initial ANLL patients which were newly diagnosed. Mononuclear cells were isolated from these samples by Ficoll. A final concentration of STI571 is 1 w M. Co-cultivate ANLLcells and STI571for 72h and then send total RNA to Shanghai BioStar Genechip limited company after it has been extracted.Results and Disscussions: There were over 730 different genes in each pair of chips, in which 156 different genes expressed together. In all different genes, 66.67%(104/156) were up-regulated, 33.33%(52/156) were down-regulated. Cellular skeleton accounted to 4.49%(7/156),oncogenes and tumor supressors 6.41%(10/156), cells receptor7.05%(ll/156), extracellular communication proteins 23.07%(36/156), cyclin proteins7.05%(l 1/156), apoptosis correlation proteins 1.92%(3/156)and other genes accounted to50% (78/156). Genes with over ten times relative difference amounted to 26.92%(42/156).It can be concluded that the mechanism of the restraint of proliferation caused by STI571 may relate to three ways: 1. STI571 realizes its biological function by affecting different signal conduction pathways through proteintyrosine kinase, guanine-nucleotide-binding protein and protein kinase C; 2. STI571 restrains the in...