The Activity Variation Of Bone Marrow Mesenchymal Stem Cells Derived From Chronic Myeloid Leukemia And The Effects Of Arsenic Trioxide On It

Objective To discuss the relationship between activity variation of bone marrow mesenchymal stem cells(BMMSCs)and the invasion of chronic myeloid leukemia(CML),by comparing the growth activities and the expression of SCF,integrin-β1 mRNA of the cultured BMMSCs derived from CML in chronic phase with control group.To study on the effects of arsenic trioxide on BMMSCs by observeing the activity variation and the change of expression of SCF \integrins-β1 mRNA of BMMSCs derived from CML after treated with different concentrations of arsenic trioxide.To provide academic and experimental foundation for futher illustrating the invasion mechanism and for exploring new clinical treatment.Methods 1,The experimental group included five CML patients, control group included five healthy persons.Observed the morphology of BMMSCs under the inverted microscopic.Counted the number of BMMSCs from day 1 to 11 after passage and portrayed the growth curve. 2,Extracted total RNA from fourth generation of BMMSCs and measured the expression of SCF\ integrins-β1 mRNA by Real-time PCR. Compared the level of two groups.3,Added Arsenic Trioxide(terminal concentrations:0μmol/L,1μmol/L,2.5μmol/L,5μmol/L)to culture system which cultured BMMSCs derived from CML.After 72h, observed the growth activities of BMMSCs under inverted microscopic. 4,Fourth generation BMMSCs derived from CML were cultured in 96-microwells plates.24h later,added arsenic trioxide(terminal concentrations of it were 1μmol/L,2.5μmol/L,5μmol/L)to culture systems.After cultured for another 48h,the inhibition of proliferation was determined by MTT assay.5,Cultured the fourth generation BMMSCs derived from CML in culture bottle for 24h,then cultured for another 48h with Arsenic Trioxide(terminal concentrations of arsenic trioxide were 0μmol/L,1μmol/L,2.5μmol/L,5μmol/L).Extracted total RNA and measured the expression of SCF\ integrins-β1 mRNA of BMMSCs by Real-time PCR.Results 1,BMMSCs in two groups mostly were shaped like long fusiform and adhered the culture bottle.BMMSCs of two groups had no difference in bioactivity and growth curve.2,The expression of SCF and integrins-β1 mRNA in BMMSCs was higher in experimental group than in control group(p<0.05).3,arsenic trioxide effected the BMMSCs(derived from CML)to adhere to the culture bottle and this effect is concentration- dependence.In 5μmol/L group,most of the BMMSCs suspended and died.4,By MTT assay,arsenic trioxide significantly inhibit the growth rate of BMMSCs.The inhibit rates were at dose dependent(p<0.05).5,arsenic trioxide could effect the expression of SFC/integrin-β1 in the BMMSCs.The expression of SCF mRNA in 1μmol/L was higher than control group,but had no statistical significance(p>0.05);The expression of SCF mRNA in 2.5μmol/L group was the highest in all groups(<0.01).The expression of SCF become lower with increasing concentration of arsenic trioxide.The expression of SCF was lower in 5μmol/L group than in control group but had no statistical significance(>0.05).The expression of integrin-β1 mRNA in 1μmol/L group was significantly higher than control group(<0.01).The expression of integrin-β1 become lower with increasing concentration of arsenic trioxide.In 5μmol/L group,the expression of integrin-β1 was lower than controal group but had no statistical significance(>0.05)Conclusion 1,The BMMSCs have no difference in growth activities between CML and control group.2,The significant high express of SCF in BMMSCs from CML can increase the proliferation of leukemia cell.And the significant high expression of integrin-β1 can improve the CML development by promoting the proliferation and decreasing the apoptosis of leukemia cells.3,Arsenic trioxide can decrease the BMMSCs adhere and proliferation and significantly incresced cells apoptosis or death.4,Low concentration of arsenic trioxide can induce BMMSCs express more SCF and integrin-β1 in vitro. The high level of SCF and integrin-β1 can promote the CML cells proliferation or inhibit them apoptosis.The expression of SCF and integrin-β1 becomes lower and the effects of arsenic trioxide on promoting apoptosis of BMMSCs become greater while increasing concentration of arsenic trioxide.So arsenic trioxide have the double functions to BMMSCs.