Clinicopathological Features And Prognosis Of Double-hit Lymphoma

Background and PurposeB-cell lymphoma with both Myc and Bcl-2 or Bcl-6 translocation is called double hit lymphoma(DHL)according to the definition in WHO(2008)classification,and will also have three Gene translocation is called Triple-hit lymphoma(THL).Another type of B-cell lymphoma was detected by immunohistochemistry with high expression of MYC and BCL-2 or BCL-6,which is called double-expression lymphoma(DEL).In recent years,many studies have sought the optimal diagnostic threshold for immunohistochemistry of MYC and BCL-2 / BCL-6 positive expression.Screening of gene translocation by immunohistochemical protein expression to identify patients who are at risk in daily practice Detection.In this study,Myc,Bcl-2,Bcl-6 translocation and expression of corresponding proteins in DLBCL were detected by FISH and IHC.The clinical and pathological features of double-hit lymphoma and double-express lymphoma were obtained,and the relationship between them was discussed and the prognosis characteristics and diagnosis and treatment were discussed.Method1.Collected data: collected from Zhengzhou University First Affiliated Hospital2010 to 2015 diagnosed with DLBCL patients with paraffin-embedded tissue specimens,the two senior lymphoma physicians in the department to re-read the diagnosis,and screened out the application of CHOP program or R-CHOP 65 patients were treated with tissue microarray(TMA)and 65 specimens were made into tissue microarray.2.Follow-up: The clinical data and pathologic data of 65 patients with DLBLC were retrospectively analyzed and the patients were followed up by telephone.3.immunofluorescence in situ hybridization: using Vysis LSI MYC two-color,rupture rearrangement probe,Vysis LSI BCL6 two-color,rupture rearrangement probe and Vysis LSI IGH / BCL2 dual fusion translocation probe(Abbott Molecular,Abbott Park,IL)The The FISH signal was analyzed using a fluorescence microscope equipped with DP72 camera and DP2-BSW software(Olympus,Tokyo,Japan)(Olympus BX51,Tokyo,Japan).4.Immunohistochemistry: The expression of MYC,BCL-2,BCL-6,CD10,MUM-1 and Ki-67 proteins were detected.5.Statistical analysis: Statistical analysis using SPSS 17.0 statistical software.Pearson chi-square test,continuity correction and Spearman rank correlation analysis are used to determine the association and correlation between variables.The survival analysis was plotted using the Kaplan-Meier curve and compared using the logarithmic rank test.Analysis of univariate and multivariate survival analysis by Cox proportional hazards regression model.When bilateral P <0.05,the results were considered statistically significant.Result1.Clinical data and follow-up: 65 patients,including 34 males and 31 females,male to female ratio of 1.1: 1,aged 4 to 81 years,mean age 50.8 years,the median age of 60 years.Patients with AnnArbor staging: I 21 cases,? stage 29 cases,? 10 cases,? 5 cases.According to the above related indicators to calculate the patient IPI,32 cases of low risk,low / moderate risk in 18 cases,10 cases of high risk,high risk in 5 cases.Thirty-six patients were treated with CHOP regimen(cyclophosphamide +doxorubicin + vincristine + prednisone)and 29 patients received rituximab(R-CHOP regimen)for 2 to 80 months 34 cases,31 cases died.2.FISH: Among the 65 cases of DLBLC,the Myc gene translocation(14/65)accounted for 21.5% of DLBCL;Bcl-2 gene translocation accounted for 18.5%(12/65);Bcl-6 gene translocation accounted for 27.7% 18/65);BCL-2 / IGH gene translocation accounted for 18.5%(12/65).Myc gene rearrangement was the only translocation in 7(10.8%)tumor samples.Six DHLs were detected.Among them,4(6.1%)had Myc and Bcl-2 gene translocation,2(3.1%)Myc and Bcl-6 gene translocation.9(13.8%)simultaneous translocation of Bcl-2 and Bcl-6.The triple translocation of Myc,Bcl-2 and Bcl-6 genes was detected in 1 case(1.5%).3.IHC:There were 16 cases of GCB in 46 cases and 46 cases of NON-GCB cases.GCB accounted for 27.6%(3/16)and NON-GCB accounted for 68.8%(11/16)in Myc gene translocation.In Bcl-2 / IgB accounted for 26.3%(6/12),NON-GCB accounted for 41.7%(5/12),GCB accounted for 33.3%(4/12),NON-GCB in Bcl-6gene translocation Accounting for 58.3%(7/12).6% of DHL patients had GCB accounting for 16.7%(1/6)and NON-GCB accounted for 83.3%(5/6).One case THL is NON-GCB.The expression of MYC protein,BCL-2 protein and BCL-6 protein in 65 cases of DLBCL were detected.33 cases(50.1%)showed high expression of MYC protein,32 cases(49.2%)showed high expression of BCL2 protein,18 cases(27.7%)showed high expression of BCL6 protein.We used ROC curve analysis to determine the percentage of the optimal cut-off value of the protein positive cells.The optimal cutoff values for MYC,BCL-2 and BCL-6 are ? 50%,? 60%(equal to the predetermined threshold)and ?20%.Myc translocation positive cases(14 cases)in13 patients with MYC protein expression,specificity of 90%.Bcl-2 translocation positive cases(12 cases)7 patients with BCL-2 protein expression,specificity of 58%.Bcl-2 translocation positive cases(18 cases)had 11 patients with high expression of BCL-62 protein,specificity of 61%.4.Prognostic analysis: Three of the six DHL patients were treated with the R-CHOP regimen and three patients treated with the CHOP regimen.Survival between the two groups was not specific.6 patients with DHL all died,the survival period of 2-19 months,the prognosis is poor.1 case of THL survived for 8 months.In contrast to MYC translocation,cleavage of the BCL-2 or BCL-6 gene did not predictOS and PFS in patients.The candidate prognostic factors were evaluated in the CHOP and R-CHOP cohorts,respectively.Survival curves showed that high expression of MYC protein was significantly associated with poor OS in CHOP and R-CHOP groups.In the CHOP cohort,BCL-2 high protein expression was significantly associated with poor OS,but not in the R-CHOP cohort.However,high expression of BCL-6 protein is a poor prognostic factor in the R-CHOP cohort,but not in the CHOP cohort.We then analyzed the prognostic effects of DHS and THS on DLBCL patients in both treatment groups.In the univariate analysis,MYC / BCL-2 protein was highly expressed,MYC / BCL-6 protein was high and MYC,BCL-2 and BCL-6 were highly expressed in CHOP and R-CHOP groups,Related.In the R-CHOP cohort,BCL-2high protein expression was only associated with poor OS when co-expressing MYC protein.BCL-2 / BCL-6 overexpression showed limited prognostic effects.The multivariate Cox regression model of IPI,immunohistochemistry(GCB versus NON-GCB),MYC,BCL-2 and BCL-6 proteins in the CHOP cohort showed IPI,high MYC protein expression and high BCL2 protein expression,MYC / BCL-2 high protein expression,MYC / BCL-6 high protein expression,BCL-2 / BCL-6 high protein expression and three high protein expression independently affect the prognosis of OS.In the R-CHOP cohort,the multivariate Cox regression model of IPI,MYC,BCL-2 and BCL-6 proteins showed high MYC protein expression,high MYC/ BCL-2 DHS,high MYC / BCL6 DHS and high THS For independent OS and PFS have independent prognostic value.High BCL2 / BCL6 DHS is also an independent prognostic value.Cox model of two groups of patients showed no high correlation between MYC protein expression and Myc gene rupture,BCL-6 protein expression showed no correlation with Bcl-6 rupture,BCL-2 high protein expression showed that IGH / Bcl-2 fusion is not relevant.The Cox model of the Myc fracture and DHS showed that the cohort in the IHC group or the FISH group model could be representative of the population of DHL as a whole.Conclusions1.DHL and DEL patients with high levels of LDH,clinical staging mostly ?-?,multiple extranodal invasion,and high IPI score of the clinicopathological features.2.For the MYC,BCL-2 and BCL-6 protein expression,the best Cut-Off values of50%,60% and 20% predicted the Myc rupture detected by FISH in DLBCL,and finally we detected the Myc The rupture was highly specific(90%),but IGH / Bcl-2fusion and Bcl-6 rupture were not found to be highly specific.3.The CHOP and R-CHOP regimens in DHL patients did not improve the prognosis of patients.