| Objective1.To study the expression of N-WASP(Neural Wiskott-Aldrich syndrome protein)in ALL and AML patients,and the correlation between expression level and clinical characteristics and prognosis,so as to provide a theoretical basis for finding new indicators to predict the prognosis of ALL and AML patients;2.To study the effects of N-WASP on proliferation and apoptosis of acute myeloid leukemia cell line(THP-1)and acute lymphoblastic leukemia cell line(Jurkat),so as to provide a theoretical basis for finding new targets for the treatment of ALL and AML.Methods1.Collectting the clinical data of ALL and AML patients diagnosed in our hospital from January 2019 to January 2021(including age,gender,blood routine results,bone marrow blast cell,peripheral blood blast cell,FAB classification,risk stratification,prognosis,etc.);According to disease status,the patients were divided into newly dignosed group,remission group and recurrence group;According to immune classification or FAB classification,ALL patients were divided into B-ALL group and T-ALL group,and AML patients were divided into M0-M7;According to the risk degree,the patients were divided into high-risk group,intermediate risk group and standard risk group;According to the median expression of N-WASP m RNA,the patients were divided into high expression group and low expression group;Taking the patients with non hematological malignant diseases in the same period and the same age group as the control,2m L bone marrow was collected respectively;2.RT-q PCR and Western Blotting were used to detect the expression of N-WASP m RNA and N-WASP protein.The relationship between the m RNA expression level and different clinical characteristics and prognosis of patients was analyzed;3.Infecting THP-1 and Jurkat cells with lentivirus to overexpress N-WASP,CCK8 was used to detect proliferation,and the expressions of JAK2,p-JAK2,STAT3 and p-STAT3 proteins in JAK2/STAT3 signaling pathway,AKT protein in PI3K/AKT signaling pathway and ERK protein in MAPK/ERK signaling pathway were detected by Western Blotting.The expression of STAT3 m RNA was detected by RT-q PCR.Results1.125 patients with acute leukemia(AL)were included in this study,including66 cases of ALL and 59 cases of AML.There were 20 cases in the control group.Among ALL patients,there were 28 males and 38 females,with a median age of 12years(1.5-70 years);There were 42 patients in newly dignosed group,14 patients in remission group and 10 patients in recurrence group;58 cases of acute B-lymphocyte leukemia(B-ALL)and 8 cases of acute T-lymphocyte leukemia(T-ALL);There were34 cases in standard risk(SR),20 cases in intermediate risk(IR)and 12 cases in high risk(HR);Among the patients with AML,there were 30 males and 29 females,with a median age of 13.5 years(1.2-69 years);There were 38 patients in newly dignosed group,17 patients in remission group and 4 patients in recurrence group;16 cases of M2,17 cases of M3,11 cases of M4,13 cases of M5 and 2 cases of M7;There were10 cases with SR,16 cases with IR and 33 cases with HR.In the control group,there were 10 males and 10 females,with a median age of 14 years(0.8-72 years).2.The expressions of N-WASP m RNA and N-WASP protein in ALL patients and AML patients were significantly lower than those in the control group(P<0.05).3.There was no correlation between the expression level of N-WASP m RNA and clinical indexes such as gender,age,blood type,blood routine results,PB,BM,hepatosplenomegaly,coagulation dysfunction,liver or kidney function damage of ALL and AML patients(P>0.05).4.For ALL patients,in terms of disease status,the expression of N-WASP m RNA in newly dignosed group and recurrence group was significantly lower than that in remission group and control group(P<0.05),and there was no difference between patients in remission and the control group(P>0.05);In terms of immunotyping,the expression of N-WASP m RNA in B-ALL and T-ALL patients was significantly lower than that in the control group(P<0.05),and the expression of N-WASP m RNA in B-ALL patients was lower than that in T-ALL patients(P<0.05);In terms of risk degree,the expression of N-WASP m RNA in high-risk group and intermediate risk group was significantly lower than that in standard risk group and control group(P<0.05),and there was no significant difference between standard risk group and control group(P>0.05);The lower the expression of N-WASP m RNA,the lower the overall survival rate(OS).5.For AML patients,In terms of disease status,the expression of N-WASP m RNA in newly dignosed group was lower than that in the control group、recurrence group and remission group(P<0.05),but there was no difference between recurrence group and remission group and the control group(P>0.05);In terms of FAB typing,there was no significant difference in the expression of N-WASP m RNA between M2、M3、M4、M5、M7 patients and the control group(P>0.05),there was also no significant difference in the expression among different types(P>0.05);In terms of risk degree,the expression of N-WASP m RNA in high risk,intermediate risk and standard risk patients was lower than that in the control group(P<0.05),and the expression of N-WASP m RNA in high-risk and intermediate risk group was lower than that in standard risk group(P<0.05).The lower the expression of N-WASP m RNA,the lower the overall survival rate(OS).6.The proliferation of THP-1 and Jurkat cell lines overexpressing N-WASP was significantly lower than the control group(P<0.05).7.Compared with the control group,there was no significant difference in the expression of Caspase-3 and Bcl-2 in THP-1 and Jurkat cell lines which overexpressing N-WASP(P>0.05).8.In THP-1 cell line overexpressing N-WASP,the expression of JAK2,p-JAK2,STAT3,p-STAT3 and AKT decreased(P<0.05),while the expression of ERK increased(P<0.05),the expression of STAT3 m RNA decreased(P<0.05);In Jurkat cell line overexpressing N-WASP,the expressions of JAK2,p-JAK2,STAT3 and p-STAT3 decreased(P<0.05),and the expressions of ERK and AKT increased(P<0.05),the expression of STAT3 m RNA decreased(P<0.05).Conclusion1.There is low expression of N-WASP gene in ALL and AML patients.Its low expression is related to disease status,risk degree and poor prognosis.It may be a new index to judge the severity and prognosis of ALL and AML patients;2.N-WASP could inhibit the proliferation of acute leukemia cell lines THP-1and Jurkat,but had no significant effect on apoptosis.3.N-WASP inhibit JAK2/STAT3 signaling pathway at the transcriptional level and affect the expression of AKT and ERK proteins.N-WASP may become a new target for the treatment of ALL and AML. |
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