The Expression And Significance Of TLR4 MRNA In Myelodysplastic Syndrome

Myelodysplastic syndrome (MDS) are clonal stem cell disorders characterized by ineffective hematopoiesis leading to peripheral blood cytopenias and by a high incidence of progression to acute myeloid leukemia (AML). Several studies have shown increased rates of programmed cell death (apoptosis) in marrow cells of patients with MDS,the most described extensive dysregulation of proapoptotic pathways, including tumor necrosis factor a (TNFa), and TNF-related apoptosis-inducing ligand(TRAIL),Fas and Fas ligand,nuclear factor-kappa B (NFκB),Caspase-3,B cell lymphoma/leukemia-2(BCI-2) etc.These factors may contributed to increase or inhibit the rates of apoptosis in bone marrow mononuclear cells of MDS patients,but the mechanism of these factors'function in the progression of MDS is already unclear.TLR4(toll-like receptor-4) is one of the TLRs family,it was important that the Toll signaling pathways play roles in the innate immunity and adaptive immunity response. Several studies have shown that the TLR4 gene can increased the rate of cell apoptosis in cancer of the stomach and neuroglia cells.It was well known that MDS exist excessive cell apoptosis in bone marrow mononuclear cells.With these factors excluded,whether there has relation between TLR4mRNA expression and MDS bone marrow mononuclear cells apoptosis? To our knowledge, this report is the first to show that TLR4mRNA expressed in the bone marrow mononuclear cells with MDS patients in our country.We investigate the expression of TLR4mRNA in de novol MDS patients bone marrow mononuclear cells and the rate of bone marrow mononuclear cells apoptosis, we studied the expression of TLR4mRNA in MDS patients bone marrow mononuclear cells whose post-treatment in 6 monthes for detect the relation between TLR4mRNA expression and prognosis of MDS patients also.Objesctives:To investigate the expression of TLR4 mRNA in bone marrow mononuclear cells of myelodysplastic syndrome patients. To analyze the relationship between TLR4mRNA expression and bone marrow mononuclear cells apoptosis of myelodysplastic syndrome patients, and the significance of TLR4mRNA expression in myelodysplastic syndrome prognosis.Methods:Real-time quantitative PCR was used to investigate the expression of TLR4 mRNA in bone marrow mononuclear cells of myelodysplastic syndrome patients and contorls . Annexin-V/FITC was used to investigate the apoptosis of myelodysplastic syndrome patients and controls.Results:1 Expression of TLR4 mRNA in de novol myelodysplastic syndrome group. The level of TLR4 mRNA expression in low risk group was higher than that in controls and in the middle and high risk group with statistical significance ( t'=-8.44,p<0.0001),(t'=7.46,p<0.0001). The level of TLR4 mRNA expression in the middle and high risk group was higher than that in controls without statistical significance(t'=-0.18,p=0.860).2 Apoptosis in myelodysplastic syndrome bone marrow mononuclear cellsThe level of apoptosis in bone marrow mononuclear cells with low risk myelodysplastic syndrome group was higher than that in controls and the middle and high risk group with statistical significanc(et'=-8.71,p<0.0001),(t'=-7.1,p<0.0001),the middle and high risk group bone marrow mononuclear cells apoptosis level without difference comparing with controls (t'=-0.96,P=0.3433).3 The level of TLR4 mRNA expression was positive relation to apoptosis in bone marrow mononuclear cells in low risk group with statistical significance r=0.654 (p=0.006),the middle and high risk group has relation to apoptosis without statistical significance r=-0.270 (p=0.422).4 The relationship between expression of TLR4mRNA and the prognosis.4.1 There were 16 cases be part of low risk myelodysplastic syndrome group,5 cases were complete remission(CR),5 cases were partial remission(PR),3 cases were hematological improved(HI),3 cases were non remission(NR) after therapied. The expression of TLR4 mRNA level in low risk myelodysplastic syndrome group which post-treatment was lower than that in de novol myelodysplastic syndrome group with statistical significance(t'=-4.73,p<0.0001). TLR4mRNA expression in bone marrow mononuclear cells with CR,PR and HI groups which post-treatment was lower than that in de novol myelodysplastic syndrome patients. ( t'=-7.61,p=0.0007 )(t'=-3.54,p=0.0122)(t'=-4.11,p=0.0274).But the TLR4mRNA expression in NR group that post-treatment without difference comparing with de novol myelodysplastic syndrome patients. (t'=-0.12,p=0.9099).4.2 There were 11 cases be part of the middle and high risk myelodysplastic syndrome group,3 cases were CR,2 cases were PR,4 cases were HI,2 cases were NR after therapied. The level of TLR4 mRNA expression in the middle and high risk myelodysplastic syndrome group which post-treatment was without difference comparing with de novol myelodysplastic syndrome patients. (t'=-0.22,p=0.8290).TLR4mRNA expression in bone marrow mononuclear cells with CR,PR,HI and NR groups which post-treatment were without difference with de novol myelodysplastic syndrome patients too( t'=0.42,p=0.7157 ),( t'=-0.33,p=0.7979 ),(t'=0.50,p=0.6357),(t'=-0.52,p=0.6937).Conclusions:1 The level of TLR4 mRNA expression in low risk myelodysplastic syndrome bone marrow mononuclear cells was significantly higher than that in controls and the middle and high risk group. expression of TLR4 mRNA in the middle and high risk group without difference comparing with controls. 2 The apoptosis level in low risk myelodysplastic syndrome bone marrow mononuclear cells was significantly higher than that in controls and the middle and high risk group,but the level of apoptosis in the middle and high risk group was without difference comparing with controls.3 There has positive relation between TLR4mRNA expression and apoptosis in bone marrow mononuclear cells with low risk group,but in the middle and high risk group,there has no relation between TLR4mRNA expression and apoptosis in bone marrow mononuclear cells.4 The expression of TLR4mRNA level in low risk myelodysplastic syndrome group which post-treatment was lower than that in de novol myelodysplastic syndrome group,TLR4mRNA expression in bone marrow mononuclear cells with CR,PR and HI groups which post-treatment was lower than that in de novol myelodysplastic syndrome patients. but the TLR4mRNA expression in NR group that post-treatment was without difference comparing with de novol myelodysplastic syndrome patients.The level of TLR4 mRNA expression in the middle and high risk myelodysplastic syndrome group which post-treatment was without difference comparing with de novol myelodysplastic syndrome patients. TLR4mRNA expression in bone marrow mononuclear cells with CR,PR,HI and NR groups which post-treatment were without difference with de novol myelodysplastic syndrome patients too.