| Objective:1.To investigate the expression of PPARD gene in acute leukemia and its relationship with the clinical data of patients,and to provide basis for future treatment and prognosis of AL;2.To investigate the mechanism of PPARD in acute lymphoblastic leukemia(ALL)and acute myeloid leukemia(AML).Methods:1.46 AML patients,31 ALL patients as the case group,and 20 non-blood disease patients as the control group were collected.Bone marrow was extracted to detect the mRNA and protein expression of PPARD.2.Culturing ALL cell lines(CEM-C1/Jurkat)and AML cell lines(THP-1/MV4-11),and detect the protein expression level of PPARD.3.CEM-C1 and Jurkat cells were treated with different concentrations of PPARD activator GW501516(0,0.1,1,5,10 μmol/L)for 24 h,48h,72h;THP-1 and MV4-11 cells were treated with different concentrations of PPARD inhibitor GSK3787(0,0.1,1,5,10μmol/L)for 24 h,48h,72h;the cell proliferation was detected by CCK-8.4.RT-q PCR and Western blotting detect the expression level of PPARD,JAK2,p-JAK2,STAT3,p-STAT3 in GW501516-treated CEM-C1/Jurkat cells,GSK3787-treated THP-1/MV4-11 cells and blank control group(untreated).5.Flow cytometry detect the changes of CEM-C1/Jurkat,THP-1/MV4-11 cell cycle and apoptosis.6.Western blotting detect the protein expression levels of CEM-C1,THP-1 cell Cyclin D1/Cyclin E1 proteins and Bcl-2/Caspase-3 proteins.Results:1.Compared with the control group,the expression of PPARD mRNA in AML was low,and the expression of protein was hight;the expression of PPARD mRNA in ALL was hight,and the expression of protein was low.2.The expression of PPARD in ALL patients with hepatosplenomegaly was higher than that without hepatosplenomegaly;and the patients with severe anemia were higher than those in the normal group and patients with mild anemia(P < 0.05).The expression of PPARD in intermediate-risk ALL patients was higher than that of the control group,low-risk group and high-risk group(P < 0.05);The expression of PPARD in B-ALL patients was higher than that of T-ALL patients in immunophenotyping,and was lower than that of the control group(P < 0.05).ALL patients with chromosomal were higher than normal patients(P < 0.05).3.The expression of PPARD in ALL patients,the percentage of bone marrow naive cells <50% was higher than bone marrow naive cells ≥50%;the high-risk group was lower than the control group(P < 0.05);M2 and M5 were lower than the control group(P <0.05).4.The expression of PPARD protein in ALL cells(CEM-C1/Jurkat)was low(P <0.05),the expression of PPARD protein in AML cells(THP-1/MV4-11)was high(P <0.05).5.CCK-8 detection results: compared with the control group,CEM-C1/Jurkat cells deal with GW501516 the OD value decrease then increase;THP-1/MV4-11 cells deal with GSK3787 the OD value decrease then increase.The optimal concentration and time of CEM-C1 cells was 1μmol/L and 48 h,the Jurkat cells was 0.1μmol/L and 24 h,the THP-1cells was 1μmol/L and 24 h,the MV4-11 cells was 1μmol/L and 48 h.6.After 1 μmol/L and 0.1 μmol/L GW501516 respectively effect CEM-C1 cells 48 h and Jurkat cells 24 h,the expression of PPARD protein was up-regulated(P < 0.05);after 1μmol/L GSK3787 respectively effect THP-1 cells 24 h and MV4-11 cells 48 h,the expression PPARD protein was down-regulated(P < 0.05).7.Compared with the control group,the expressions of p-JAK2,STAT3,p-STAT3 in CEM-C1 and Jurkat cells were all decreased(P < 0.05),the expressions of JAK2,p-JAK2,STAT3,p-STAT3 in THP-1 and MV4-11 cells were all decreased(P < 0.05).8.The results of cell cycle show that: comparing with the control group,the percentage of G0/G1 phase in CEM-C1,Jurkat cells wree increased(P < 0.05),the percentage of G0/G1 phase in THP-1,MV4-11 cells wree increased(P < 0.05).9.The results of apoptosis show that: up-regulation of PPARD expression had no effect the apoptosis rate of CEM-C1 and Jurkat cells(P > 0.05);down-regulation of PPARD expression also had no effect the apoptosis of THP-1 and MV4-11 cells(P > 0.05).10.Compared with the control group,the expressions of Cyclin D1,Cyclin E1 proteins in CEM-C1 and THP-1 cells were decreased(P < 0.05).11.Compared with the control group,the expressions of Bcl-2,Caspase-3 proteins in CEM-C1 and THP-1 cells was no difference(P < 0.05).Conclusion:1.ALL patients have low expression of PPARD protein,while AML patients have high expression of PPARD protein.2.PPARD expression is associated with risk and FAB classification in AL patients.3.ALL cells have low expression of PPARD protein and GW501516 can inhibit ALL cell proliferation,AML cells have high expression of PPARD protein,and GSK3787 can inhibit AML proliferation.4.PPARD may inhibit the proliferation of ALL and AML cells by affecting the JAK2/STAT3 signaling pathway.5.May inhibit the proliferation of ALL and AML cells by affecting the cell cycle through the expression of Cyclin D1 and Cyclin E1 proteins.6.PPARD has nothing to do with apoptosis of ALL and AML cells. |
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