| Objective: The aim of the study to explore the relationships between expressionlevel of the hIL-3receptorsα isoforms SP1and SP2and the clinical characteristics,and serum IL-3levels in patients with acute myelogenous leukemia. We also wantedto assess the the effects of7G3, which is the specific antibody to the SP1, on theproliferation and differentiation of the primary leukemia cells in vitro culture.Methods:1．Use real-time PCR to detect the relative quantification of hIL-3RαSP1and SP2transcript on the mononuclear cells from77acute myelogenousleukemia patients’ bone marrow or blood, and sorted CD34+AML cells from11acuteleukemia patients’ bone marrow or blood, with sorted CD34+cells from14normalumbilical cord blood as a control.2. Western blot was used to identify the expressionof SP1.3. ELISA test was used to measure the concentration of serum IL-3in patientswith acute leukemia, with14normal people serum as control.4. With establishing aprimary culture of leukemia cells, assessing the effect of7G3, which is the specificantibody to the SP1, on the the proliferation and differentiation of IL-3dependentprimary leukemia cells by Brdu and Wright-Giemsa assays.5. Statistic analysis of thedata was performed using the SPSS17.0software. Spearman rank test was used toevaluate the bivariate correlations. Kruskal-Waillis analysis was used to explore themultivariate correlations. The χ2test was used to identify complete remission(CR).For the survival analysis the Kaplan-Meier method was used. And the log-rank testwas used to check for overall survival. Dimension reduction-optimal scaling levelanalysis was used to evaluate multivariates analysis. Diffenrences were consideredsignificant if P-values were below0.05.Results:1. The expression of SP1on AML CD34+cells was higher than that of the normal CD34+cells (P=0.0109)，and higher than the unsorted AML cells as well(P=0.0243). However we didn’t get the same results from SP2.2. The overexpression of SP1in AML(non-APL) correlated with higher WBCand higher blast ratio, and predicted a lower overall survival （P=0.0307）. Theoverexpression of SP2correlated with higher WBC(r=0.235， P=0.039) and higherblast ratio (r=0.228，P=0.047). However, this correlation was not seen with prognosisprediction.3. The proliferation of the primary leukemia cells which had a good reactivity toIL-3declined to some extent when7G3blocked hIL-3Rα SP1, and the proliferationof the week reactive ones didn’t have significant change. No obvious change wasobserved on the cell morphology after SP1was blocked.4. A nonlinear relationships existed between the autonomous growth potential ofleukemia blast cells and the response to the exogenous IL-3, that the sample withhigher autonomous growth had a lower response to the exogenous IL-3, the samplewith lower antonomous growth had no correlation with the high response to IL-3. Anonlinear relationships existed between the serum IL-3levels and the ratio of CR, thatamong the patients whose serum IL-3level was more than the median of normalpeople, a higher serum IL-3level predicted a lower CR ratio, however, among thepatients whose serum IL-3level was less than the median of normal people, nosignificant correlation was exist between the serum IL-3level and CR ratio. Anonlinear relationships existed between the autonomous growth potential of leukemiablast cells and the ratio of CR, that among the samples whose autonomous growthwas higher than the median level, a higher autonomous may predict a low CR ratio,however among the samples whose autonomous growth was lower than the medianlevel, no significant relation was exist between autonomous growth potential and CRratio.Conclusion: AML CD34+cells had a higher expression of SP1than the normalCD34+cell group, also higher than the AML unsorted leukemia blast cells. And theexpression of SP1can be served as an indicator for patients’ prognosis. |
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