TGF-β1 Affected The Proliferation Of CMPD Patient Bone Marrow Fibroblasts And Its Expression Of Smad2 And Smad7 MRNA

Objective By comparing TGF-β1 of different concentration affected the proliferation of bone marrow fibroblasts and its expresssion of Smad2 and Smad7 mRNA, and the difference between IDA and CMPD Patient's bone marrow fibroblasts, to understand the mechanism of myelofibrosis. Methods bone marrow fibroblasts were separated from 2 normal human and 2 CMPD patients. The culture medium consisted DMEM of lesser glucose and 10% fetal calf serum(FCS). Cells were incubated at 37°C, 5% C02 condition, digest the bone marrow fibroblasts with 0.25% trypsin and 0.02% EDTA. The proliferation of normal human and CMPD patient's bone marrow fibroblasts affected by 0.01,0.1,1,2,5ng/ml TGF-β1 was measured by MTT. The mRNA expression of Smad2 and Smad7 of nomal human and CMPD patient's bone marrow fibroblasts stimulated by 0.01,0.1,1,5ng/ml TGF-β1 was measured by RT-PCR. Results MTT showed that TGF-β1 can stimulate the growth of marrow fibroblasts, especially at the 2ng/ml and 5ng/ml(P<0. 05). There was no difference between normal human and CMPD patient's bone marrow fibroblasts proliferated by the same concentration of TGF-β1. RT-PCRshowed that the mRNA expression of Smad2 and Smad7 up-regulated when increased the concentration of TGF-β1, but there was no difference between normal human and CMPD patient's bone marrow fibroblasts stimulated with the same concentration of TGF-β1. Conclusion some concentration of TGF-β1 can stimulate the proliferation of bone marrow fibroblasts. TGF-β1 can activate the Smads pathway in human bone marrow fibroblast, and can increase the mRNA expression of Smad2 and Smad7. There was no abnormity of CMPD patient's bone marrow fibroblasts in the TGF-β1/Smad pathway.