| Objective: To detect the polymorphism of RPMS1 gene in peripheral blood EBV-positive samples from patients of leukemia,myelodysplastic syndrome and lymphoma in Qingdao,Shandong Province.The aim of this study was to understand the variation characteristics of RPMS1 gene in EBV-positive patients with malignant hematological diseases in Qingdao,and to explore the relationship between RPMS1 gene mutation and the development of leukemia,myelodysplastic syndrome and lymphoma.Methods: A total of 891 patients with malignant hematological diseases and 93 healthy adults in Qingdao,Shandong Province were selected as subjects.Detection of EBV BamHⅠ-W fragments in peripheral blood of leukemia,MDS and lymphoma patients and throat washing samples of healthy adults by PCR to screen EBV positive specimens.Nested-PCR and DNA sequencing were used to analyze the gene sequences of RPMS1 from218 cases of EBV-positive samples of the patients’ peripheral blood with malignant hematopathy in Qingdao.The distribution of RPMS1 variants in malignant hematopathy cases was compared with the data from healthy donors in the same area.Result:(1)A total of 391 EBV-positive specimens were detected from leukemia,myelodysplastic syndrome,lymphoma and throat washing samples of healthy human.Among them,137 cases(30.38%)were detected in leukemia specimens,56 cases(32.56%)were detected in myelodysplastic syndrome specimens,114 cases(42.54%)were detected in lymphoma specimens and 84 cases(90.32%)were detected in throat washing samples of healthy human.(2)Five subtypes of RPMS1 gene,RPMS1-A,RPMS1-B2,RPMS1-C,RPMS1-E and RPMS1-F,were observed in 391 isolates.(3)Among the five RPMS1 subtypes,RPMS1-A was the main subtype,which was detected in 281 specimens.Its gene sequence was highly conserved,and it was basically consistent with the standard strain B95-8 except for a few scattered mutations.The ratio of RPMS1-A type in throat washing samples of healthy people was significantly higher than that in acute myeloid leukemia,Hodgkin lymphoma and non-Hodgkin lymphoma(P<0.05),but there was no significant difference with acute lymphocytic leukemia,chronic leukemia and myelodysplastic syndrome(P>0.05).And there was no significant difference in the distribution of RPMS1-A subtypes among different hematological malignancies(P>0.05).(4)RPMS1-B2 wascharacterized by P50 L mutation.The ratio in throat washing samples of healthy people was significantly higher than that in acute myeloid leukemia,chronic leukemia,myelodysplastic syndrome,Hodgkin’s lymphoma and non-Hodgkin’s lymphoma(P<0.05).However,there was no significant difference with acute lymphoblastic leukemia(P>0.05).And there was no significant difference in the distribution of RPMS1-B2 subtypes among different hematological malignancies(P>0.05).(5)RPMS1-C was characterized by D51 N mutation.The ratio in throat washing samples of healthy people was significantly lower than that of acute myeloid leukemia(P<0.05),and there was no significant difference with the other five malignant hematological diseases(P>0.05).The ratio of RPMS1-C subtype in acute myeloid leukemia was significantly higher than that in acute lymphoblastic leukemia,chronic leukemia,myelodysplastic syndrome,Hodgkin lymphoma and non-Hodgkin lymphoma(P<0.05),and the ratio of RPMS1-C subtype in non-Hodgkin lymphoma was significantly higher than that in myelodysplastic syndrome(P>0.05).(6)RPMS1-E subtypes were divided into RPMS1-E1 and RPMS1-E2,characterized by S65 N and T85 R mutations,respectively.The ratio in throat washing samples of healthy people was significantly lower than that of chronic leukemia and non-Hodgkin lymphoma(P<0.05),and there was no significant difference with the other four malignant hematological diseases.The rate of RPMS1-E subtype in chronic leukemia was significantly higher than that in acute lymphoblastic leukemia,acute myeloid leukemia and myelodysplastic syndrome(P<0.05).(7)RPMS1-F subtypes were divided into RPMS1-F1 and RPMS1-F2,which were characterized by S98 G mutation and S101 I mutation,respectively.The ratio of RPMS1-F1 in throat washing samples of healthy people was significantly lower than that in acute lymphoblastic leukemia(P<0.05),and there was no significant difference with the other five malignant hematological diseases(P>0.05).While The ratio of RPMS1-F2 in throat washing samples of healthy people was significantly lower than that of chronic leukemia,myelodysplastic syndrome,Hodgkin’s lymphoma and non-Hodgkin’s lymphoma(P<0.05),and there was no significant difference with acute lymphoblastic leukemia and acute myeloid leukemia(P>0.05).The ratio of RPMS1-F2 in acute lymphoblastic leukemia was significantly lower than that in chronic leukemia,myelodysplastic syndrome,Hodgkin’s lymphoma and non-Hodgkin’s lymphoma(P<0.05).The ratio of RPMS1-F2 in Hodgkin’s lymphoma was significantly higher than that in acute myeloid leukemia,chronic leukemia,myelodysplastic syndrome and non-Hodgkin’s lymphoma(P<0.05).Conclusion:(1)RPMS1-A was the main RPMS1 subtype in EBV-postive malignant hematopathy and healthy donors from Qingdao,which was basically consistent withstandard strain B95-8 except for sporadic mutations.(2)RPMS1-B2 and RPMS1-C were characterized by P50 L mutation and D51 N mutation,respectively.RPMS1-C may be related to the occurrence of.(3)RPMS1-E was the least,characterized by S65 N mutation and T85 R mutation.This type may be related to the occurrenceof chronic leukemia.(4)RPMS1-F genotype was characterized by S98 G mutation and S101 I mutation,which may be related to the high risk of Hodgkin’s lymphoma. |
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