| Objective:1?To detect the incidence of IL-7R,NOTCH1,FBXW7,WT1 and PHF6 in 125patients with T cells acute lymphocytic leukemia(T-ALL)in China;and to explore the mutation site of IL-7R in patients with T-ALL.We used gene sequence analysis to detect the mutation sites and types of IL-7R,NOTCH1,FBXW7,WT1 and PHF6.We explored the features and clinical significance of IL-7R,NOTCH1,FBXW7,WT1 and PHF6 in patients with T-ALL.2?To probe the incidence and clinical significance of constitutional pericentric inversion of chromosome 9 in various kinds of diseases of the blood system.To study the condition of fusion gene in patients with inversion of chromosome 9 in diseases of the blood system.To compare the condition of patients with inversion of chromosome 9 and those without inversion of chromosome 9 after accepting hematopoietic stem cell transplantation.Methods:First Part Construction We collected 125 T-ALL patients'bone marrow or peripheral blood cells.All cases were studied by R-band karyotypic analysis using direct method and/or short-term culture for chromosomes preparation,PCR were performed to detect the IL-7R gene mutation.We used DNA extraction kit to extract genomic DNA,and used polymerase reaction to amplify genomic DNA.Then we used two-way sequencing method to detect condition of IL-7R in the application of PCR product of 125 patients with T-ALL.Second Part(1)Karyotype analysis:We collected 5 ml heparin anticoagulation in patients with peripheral blood,and cultured 72 hours after conventional preparation of chromosome.All cases were studied by R-band karyotypic analysis using direct method and/or short-term culture for chromosomes preparation,PCR were performed to detect the karyotype analysis.The description of abnormal karyotype was used by international naming system of human genetics 2013.(2)We used PCR technique to detect the fusion gene of PML,bcr-abl,AML-ETO,EVI1,CBF beta-MYH11,MLL-AF6,AML-AF4,SET-NUM214,SIL-TALI,IgH rearrangement,TCR rearrangement and BCL1-IgH,and so on.Results:First Part(1)The mutation rates of IL-7R in T-ALL was 5.60%.Seven leukemias with IL-7R mutations exclusively affected exon 6,without exon 5.There are 6 adults and 1child,4 men and 3 women in IL-7R gene mutations.All seven cases had an in-frame insertion or substitution at residues P240–S246 of the IL7R transmembrane domain.2/7samples had NOTCH1 mutations and 1/7 samples had PHF6 mutation.(2)There were statistically significant between patients with FBXW7 and PHF6 gene mutations and without in WBC.There were statistically significant between patients with WT1 and PHF6 mutations without in LDH.There were statistically significant between patients with IL-7R gene mutations and without in bone marrow blasts cell,and there were no statistically significant in other indicators.(3)The rate of IL-7R,NOTCH1,FBXW7,WT1 and PHF6 mutation in 125 T-ALL patients respectively were 5.60%,44.0%,9.60%,3.20%and 12.80%.In 125 patients with T-ALL,the number of ALL-1,ALL-2,ALL-3 respectively were 42,52 and 31.The rate of IL-7R,NOTCH1,FBXW7,WT1 and PHF6 mutation in ALL-1 patients respectively were4.76%,42.9%,7.14%,2.38%and 16.67%.The rate of IL-7R,NOTCH1,FBXW7,WT1and PHF6 mutation in ALL-2 patients respectively were 3.85%,44.2%,13.5%,1.92%and11.54%.The rate of IL-7R,NOTCH1,FBXW7,WT1 and PHF6 mutation in ALL-1patients respectively were 9.68%,45.20%,6.45%,9.68%and 9.68%.The rate of IL-7R,NOTCH1,FBXW7,WT1 and PHF6 mutation in 69 cases with normal karyotype were5.80%,46.40%,11.59%,4.35%and 13.04%,respectively.The rate of IL-7R,NOTCH1,FBXW7,WT1 and PHF6 mutation in 56 cases with abnormal karyotype of chromosome were 5.36%,41.1%,7.14%,3.57%and 12.50%,respectively.Second Part(1)The total incidence of inversion of chromosome 9 in the diseases of the blood system was 1.44%.The incidence of inv(9)in patients with acute myelogenous leukemia and without acute myelogenous leukemia were 3.28%(172/5251)and 1.18%(399/34338),respectively;there were statistical significance between the two groups(p<0.01).The incidence of inv(9)in patients with aplastic anemia and without aplastic anemia were 3.64%(43/1181)and 1.39%(528/38408),respectively;there were statistical significance between the two groups(p<0.01).(2)PML-RAR alpha,BAR-ABL,AML-ETO,EVI1,CBF beta-MYH11,MLL-AF6,AML-AF4,SET-NUM214,SIL-TALI,IgH rearrangement and TCR rearrangement,BCL1-IgH and other fusion gene were detected in patients with inv(9).(3)The absolute neutrophil count(ANC)>0.5*10~9/l in patients with inv(9)who accepted hematopoietic stem cell transplantation was in 12th days after transplantation,and PLT>20*10~9/lwas16thdaysaftertransplantation.Theabsoluteneutrophil count(ANC)>0.5*10~9/l in patients without inv(9)who accepted hematopoietic stem cell transplantation was in 12th days after transplantation,and PLT>20*10~9/l was 13th days after transplantation.Conclusion:1?IL-7R mutation might be have an important role in the occurrence and development of ALL.IL-7R mutation might be haven a certain relationship in the pathogenesis of T-ALL.NOTCH1,FBXW7,WT1 and PHF6 gene mutations might also have relationships with T-ALL.2?There were no statistical significance differences between patients with IL-7R,NOTCH1,FBXW7,WT1 and PHF6 mutations and without in gender,subtypes of T-ALL,cytogenetic classification.3?IL-7R,Nocth1,FBXW7,WT1 and PHF6 mutation molecular targets to make new targeted therapy drugs become possible.Aimed at IL-7R,Nocth1,FBXW7,WT1 and PHF6 mutation new targeted therapy has broad prospects in clinical application.4?The inversion of chromosome 9 can occur in various kinds of diseases of the blood system.The incidence of inv(9)in the patients with acute myelogenous leukemia and aplastic anemia respectively were 3.28%and 3.64%,which were higher than those in common people.The incidence of inv(9)in the other patients were similar with common people.5?The time of achieving ANC recovery in patients with inv(9)and without inv(9)after hematopoietic stem cell transplantation were almost similar,while the time of achieving platelet count recovery in patients with inv(9)was longer than those without inv(9). |
PDF Full Text Request