Detection Of Bcl-2/IgH And IgH Rearrangements In Peripheral And Bone Marrow Lymphocytes Of The Patients With Non-Hodgkin Lymphoma

Objective:To observe the Bcl-2/IgH and IgH gene rearrangements in peripheral and bone marrow lymphocytes of the patients with non-Hodgkin lymphoma (NHL), then explore the relationship of Bcl-2/IgH, IgH gene rearrangements with early diagnosis and evaluate with therapy effect of NHL.Methods:DNA was extracted from paired peripheral blood (PB) lymphocytes and bone marrow (BM) lymphocytes in 70 B-NHL (included 30 DLBCL,15 small B-cell lymphoma,8 chronic lymphocytic leukemia,3 MALT lymphoma,2 Burkitt lymphoma,1 Marginal zone lymphoma,1 Mantle cell lymphoma,10 T- NHL),7 lymphonode inflammation and 20 healthy controls. The Bcl-2/IgH, IgH gene rearrangments were assayed by polymerase chain reaction (PCR) and compared with results of pathological biopsy and evaluated with therapy effect. B lymphoma cell line, Raji was used as positive control. The expression of Bcl-2/IgH, IgH gene rearrangments were measured with semi-quantitive PCR. We discussed the relationship of Bcl-2/IgH, IgH gene rearrangements with clinical manifestation (lymphoma stage, systemic symptoms,β2-MG level, LDH level, bone marrow involvement, liver and spleen infiltration) and analyzed by SPSS 13.0 statistics software.Results:1. Bcl-2/IgH gene rearrangement from peripheral blood lymphocytes (36.7%) and bone marrow lymphocytes (33.3%) in 30 diffused large B-cell lymphoma (DLBCL) was seen more frequently than the other B-NHL cases (6.7%), T-NHL cases (0%), lymphonode inflammary cases (0%) and 20 healthy controls (5%) respectively (p<0.05).2. The Bcl-2/IgH gene rearrangement was not significantly different from peripheral blood lymphocytes or bone marrow's (36.7% & 33.3%) (p>0.05).3. The quantity of rearranged Bcl-2/IgH gene of 8 DLBCL cases obviously reduced from 0.53 to 0.28 (p<0.05) after 2 cycles of R-CHOP chemotherapy and totally disappeared after 6 cycles of R-CHOP chemotherapy.4. More patients (81.8%) with Bcl-2/IgH gene had higher level of serum LDH, while did less the patients (28.6%) without this gene rearrangement (p<0.05). None of lymphoma stage, systemic symptoms,β2-MG level, bone marrow involvement, liver and spleen infiltration was significantly correlated with Bcl-2/IgH gene rearrangement quanlity.5. In comparision, the results of the IgH gene rearrangment of 20 DLBCL cases (45%) and the other B-NHL cases (newly diagnosed or relapsed patients, DLBCL was excluded) (46.7%) were not significantly different (p>0.05). But 20 healthy persons, 10 T-NHL cases and 7 lymphonode inflammary cases were all negative (p<0.05).6. The quantity of rearranged IgH gene of 7 DLBCL cases obviously reduced from 0.42 to 0.13 after one course of R-CHOP chemotherapy (p<0.05) and totally disappeared after 2 courses of R-CHOP chemotherapy.7. The IgH gene rearrangement was not significantly different from peripheral blood lymphocytes or bone marrow's (43.3% & 36.7%) (p>0.05). But the results of semi-quantitive PCR from bone marrow were higher than that from peripheral blood.8. More patients (90%) with IgH gene rearrangement had higher serum LDH, while did less the patients (30%) without this gene rearrangement (p<0.05). None of lymphoma stage, systemic symptoms,β2-MG level, bone marrow involvement, liver and spleen infiltration was significantly correlated with IgH gene rearrangement quanlity.Conclusions:1. The Bcl-2/IgH gene rearrangement might be used as a specific indicator for improving DLBCL early diagnosis and accurating evaluation of the chemotherapy effects and corrected with LDH level.2. The IgH gene rearrangement might be used as specific indicator for early diagnosis and evaluation of efficacy in B-NHL and corrected with LDH level.