The MRNA Expression Associated With RNA M~6A Methylation Modification In Bone Marrow Mononuclear Cells From Acute Leukemia Patients And Its Clinical Significance

[Objective]To investigate the METTL3?METTL14?ALKBH5?FTO?WTAP mRNA expression which associated with RNA m6A Methylation in Bone Marrow Mononuclear Cells(BMMCs)from Acute Leukemia patients and Its Clinical Significance[Method]From Jul 2015 to Dec 2018,147 patients newly diagnosed acute leukemia from the First Affiliated Hospital of Soochow University were enrolled the study,including AML 81cases,B-ALL 40 cases,T-ALL 24 cases,HAL 2 cases and 19 healthy controls.Mononuclear cells were obtained by centrifugation on Ficoll-hypaque medium,METTL3,METTL14,ALKBH5,FTO and WTAP-mRNA in BMMCs were detected by real-time PCR.The relationships among METTL3,METTL14,ALKBH5,FTO and WTAP mRNA expression,clinical characteristics and prognosis of patients were analyzed.[result]Among 74 non-M3 AML patients,there were 22 patients with NPM1 mutation,31 patients with FLT3-ITD mutation,4 patients with IDH1 mutation,19 patients with DNMT3 A mutation,4 patients with TP53 mutation,6 patients with ASXL1 mutation,13 patients with NRAS mutation,18 patients with CEBP? double mutation,6 patients with EZH2 mutation,5 patients with c-Kit mutation,10 patients with TET2 mutation,10 patients with GATA2 mutation and 6 patients with RUNX1 mutation.After first course induction chemotherapy,50 patients were achieved complete remission(CR),4 patients were achieved partial remission(PR)and 15 patients were achieved non-remission(NR).After the second course induction chemotherapy,14 patients were achieved CR,2 patients were achieved PR and 3 patients were no respond.Among 40 B-ALL patients,18 patients were involved in BCR-ABL fusion genes,where RUNX1 mutation in 2 cases,FLT3-ITD mutation in 4 cases,TP53 mutation in 3 cases,NOTCH1 mutation in 1 case,NRAS mutation in 3 cases,PHF6 mutation in 1 case,BCOR mutation in 4 cases,PAX5 mutation in 3 cases.After first course induction chemotherapy,33 patients were achieved CR and 6 patients were no respond.After the second course induction chemotherapy,4 patients were achieved CR.There were 24 patients with T-ALL including 3 with SIL-TAL1 fusion gene,2 with SET-NUP214 fusion gene,4 with PHF mutation,7 with NOTCH1 mutation and 3 with RUNX1 mutation.After first course induction chemotherapy,20 patients were achieved CR and 3 patients were no respond.the relative mRNA expression of METTL3,METTL14 and WTAP in acute leukemia group was significantly higher than that in healthy control group(p<0.001),but the relative mRNA expression of FTO gene in the disease group was lower than that in the healthy control group(p<0.05).There was no significant difference in the mRNA expression of ALKBH5 gene between the disease group and the control group.Compared with ALL group,the expression of METTL3 and WTAP genes was significantly higher in AML group(p<0.0001),In acute lymphoblastic leukemia group,the expression levels of FTO and WTAP genes in T-ALL group were significantly higher than those in B-ALL group(all P<0.05).We analyzed the correlation between the mRNA expression of METTL3,METTL14,ALKBH5,FTO,WTAP and clinical characteristics.Among 74 AML patients without M3,the mRNA expression of METTL3,METTL14,ALKBH5,FTO,WTAP were not significantly correlated with age,sex,leukocyte count,platelet count and chromosome karyotype at first diagnosis(p>0.05).The level of WTAP mRNA expression was positively correlated with WT1 expression(r=0.681,P<0.0001).In addition,METTL3 gene expression in NPM1+group was higher than that in negative group(p<0.05),WTAP gene expression in FLT3-ITD+group was significantly higher than that in negative group(p<0.05),and WTAP gene expression in CEBPa double mutation group was significantly lower than that in non-mutation group(p<0.05).In B-ALL group,the expression level of METTL3 gene was significantly higher in high leukocyte group than in low leukocyte group(p<0.05);WTAP mRNA expression level was significantly decreased in BCR-ABL fusion gene positive patients,but significantly increased in WT1 positive patients(P<0.05),and the METTL3 mRNA expression in hyperleukocyte group was significantly higher(P<0.05).The mRNA expression of METTL3,METTL14,ALKBH5,FTO,WTAP in T-ALL group was no significant correlation with clinical and biological prognostic factors(p>0.05).Among all patients with acute leukemia receiving standard induction chemotherapy,the mRNA expression levels of METTL3,METTL14,ALKBH5,FTO and WTAP genes were not significantly correlated with the efficacy of treatment,and METTL3,METTL14,ALKBH5 and FTO genes was not correlated with overall survival(OS),disease-free survival(DFS)and cumulative relapse rate.In B-ALL alone,the DFS rate decreased significantly(p<0.05)and the cumulative relapse rate increased significantly(p<0.05)[Conclusion]Except for ALKBH5,METTL3?METTL14??FTO and WTAP mRNA expression in BMMCs from acute Leukemia patients were different from healthy control.The mRNA expressions of METTL3,METTL14,FTO and WTAP in different types of acute leukemia were also different.Clinical characteristics and prognosis of acute Leukemia patients may be closely related to RNA m6A methylation modification.