Parthenolide Intervenes The Jurkat Cells Adhension To The Bone Marrow Stromal Cells And Its Possible Mechanism

Objective:Through the simulation of the bone marrow microenvironment in vitro,we aim to investigate the effect of parthenolide intervenes the Jurkat cells adhesion to BMSCs which from ALL patients,and explore its possible mechanism. Methods:To construct the co-culture model by Jurkat cells and BMSCs of patient with ALL,we chose 6 completely response patients who were diagnosed as acute lymphoblastic leukemia by FAB classification and MICM classification,which from October 2013 to March 2014 in Affiliated Hospital of Guilin Medical College. We cocultured the Jurkat cells and BMSCs which were isolated from those patients’ posterior spinet fluid. Several groups were setted:Jurkat cells group,BMSCs group,BMSCs+Jurkat cells group,BMSCs+Jurkat cells+ PTL4 μmol /L group,BMSCs+Jurkat cells+ PTL8 μmol/L group,BMSCs+Jurkat cells+ PTL12 μmol /L group,BMSCs+Jurkat cells+PTL16 μmol /L group. The adhesion rate of the Jurkat cells adhesion to BMSCs and intervention effect of parthenolide at 24 and 48 hour were detected by MTT;ELISA was performed to test the expression of s VCAM-1 in co-cultured cells supernatant and intervention effect of parthenolide; RT-PCR was performed to test the expression of VCAM-1 m RNA in BMSCs. Results:The results of MTT shows that the adhesion rate for PTL of 4μmol/L, 8μmol/L, 12μmol/L,16μmol/L at 24 h were 39.10%±3.80%,28.52%±4.74%,21.01%±3.55%,13.92%±1.88%,all significantly lower than the control group59.64%±4.32%(F=133.21,P<0.01),the significant difference was found by comparing any two concentration groups(P<0.01).the adhesion rate for PTL of4μmol/L,8μmol/L,12μmol/L,16μmol/L at 48 h were 30.21%±0.49%,22.43%±1.76%, 14.71%±0.65%, 8.00%±1.19%,all significantly lower than the control group67.37%±5.02%(F=533.70,P<0.01),the significant difference was found by comparing any two concentration groups(P<0.01). ELISA assay shows that the expression of s VCAM-1 in cocultured cells supernatant in PTL was lower than the control group(F=545.56,P<0.01),PTL inhibited BMSCs secreting s VCAM-1 by dose-dependence(R=-0.99,P<0.01). In PTL16μmol/L group,its concentration level of s VCAM-1 decreased to the BMSCs group,there was no significant differences beteween these two groups(P>0.05).RT-PCR shows that the relative expression quantity of VCAM-1 m RNA in BMSCs was lower than the control group(F=7006.15,P<0.01),PTL inhibited the expression of BMSCs VCAM-1 m RNA by dose-dependence(R=-0.99,P<0.01).In PTL16μmol/L group,its expression of VCAM-1 m RNA decreased to the BMSCs group,there was no significant differences beteween these two groups(P>0.05). It was in accord with ELISA. Conclusion:PTL inhibited Jurkat cells adhesion to BMSCs in dose-dependence in vitro,its mechanism may be associated with the inhibition of expression of VCAM-1.