| Objective: DLBCL is a kind of independent disease,it has obvious heterogeneity in clinical manifestations,pathological characteristics,molecular subtypes,immunophenotyping and clinical prognosis.According to the gene expression profile(GEP),DLBCL was divided into GCB subtype(GCB-DLBCL),ABC subtype(ABC-DLBCL),and the third subtype that about 15%-30% of patients could not be classified.The expression of CD10,Bcl-6 and MUM1 were detected by immunohistochemistry,according to their expression results,DLBCL was divided into GCB and non GCB subtypes(non-GCB-DLBCL).Although in the era of rituximab,R-CHOP significantly improved the prognosis of patients with DLBCL,the prognosis of patients with ABC-DLBCL was still poor.The 5-year overall survival rate(OS)of DLBCL was only 60-70%,GCB-DLBCL was about 75%,and ABC-DLBCL was about35%.There are still 40%-50% of the patients are not sensitive to the standard treatment plan,and the mechanism is not clear.The median survival time of these patients was only 4 months,and the median progression free survival time was only 3months.Therefore,it is of great significance for the early diagnosis and treatment of DLBCL,the improvement of curative effect and the improvement of prognosis to understand the pathogenic mechanism of DLBCL and to find the specific molecular targets for the early diagnosis,curative effect and prognosis evaluation.With the rapid development of science and technology,the second generation sequencing technology(NGS)can realize the whole genome,the whole exon,the transcriptome sequencing,and determine the recurrent gene mutation in DLBCL.Different subtypes involve different gene mutations,such as EZH2,GNA13 and Bcl-2mutations common in gcb-dlbcl,while ABC subtypes mainly involve gene mutations related to BCR / NF-κB signaling pathway,in which My D88 and CD79 B are the two key genes in the signaling pathway and also the current hot genes.According to a recent report in the New England Journal,574 patients with DLBCL were analyzedretrospectively.According to the different genotypes,epigenetics and clinical features of DLBCL,DLBCL was divided into four subtypes at the molecular level: MCD,BN2,N1 and EZB.The main features of MCD were My D88 and CD79 B mutations.These mutations promote the abnormal activation of NF-κB anti apoptotic signaling pathway and B cell receptor(BCR)signaling pathway,and block cell apoptosis,thus promoting tumor growth.Among them,CD79 B mutation can increase the expression of BCR on the surface of lymphocytes and make the negative control signal of BCR fail,which can lead to the chronic and sustained activation of NF-κB signal pathway,while My D88 l265 P mutation can enhance the activity of IL-1 receptor-related kinase(IRAK1,4)and activate NF-κB signal transmission.Recently,it was reported that about38.7%-94% of DLBCL highly expressed My D88,which was not related to the mutation of My D88 l265 P.Therefore,there may be other mechanisms regulating the expression of My D88 in DLBCL.Micro RNA(micro RNA,mi RNA)is a small noncoding RNA with a length of about 18-25 nucleotides.As an important epigenetic regulatory factor,mi RNA exists widely in the biological community,mainly through binding with the 3 ’-noncoding region(3’-UTR)of the target gene m RNA,resulting in m RNA degradation or translation inhibition,and plays an epigenetic modification role at the post transcriptional level in cell differentiation,development,proliferation,apoptosis and other stages.mi R-525-5p,as a newly reported micro RNA,has the function of promoting and inhibiting cancer in different tumors.In the early stage of our research group,mi RNA microarray technology was used to detect the down-regulation of mi R-525-5p expression in DLBCL tissue.Subsequently,we found that mi R-525-5p combined with 3’untranslated region(3’UTR)target of My D88 in the database prediction of targetscan 7.0 and mirwalk 3.0.At present,the relationship between mi R-525-5p and My D88 has not been reported.The role of mi R-525-5p and My D88 in DLBCL is not clear.Based on the previous research results of the research group,this study combines databaseinformation retrieval mining and molecular biology experiments.The first part is to detect the expression level of mi R-525-5p and My D88 protein in DLBCL tissue,analyze the relationship between the two and their clinicopathological significance.The second part is to detect the mutation level of My D88 l265 P and CD79 B,and analyze the relationship between the mutation and NF-κB signal activation,so as to find a new target for DLBCL.Mthods: This study reviewed and screened the cases diagnosed as DLBCL and lymph node reactive hyperplasia(RH)in the pathology department of the First Affiliated Hospital of Dali University from 2008 to 2019.The first part:(1)mi RWalk3.0 and Targetscan database predict the binding target gene of mi R-525-5p and target mi RNA of My D88;(2)IHC detect the expression of My D88 protein;(3)q PCR detect the expression of mi R-525-5p.The second part:(1)IHC detected NF-κB / p65 protein;(2)PCR amplification and sequencing of exon 5 of My D88 and CD79 B.SPSS17.0 was used for statistical analysis results(p < 0.05).Results: The first part:(1)The binding site of mi R-525-5p in 3’UTR of My D88 gene was retrieved by mi RWalk 3.0 and Targetscan database;(2)The high expression of My D88 in 71.4%(55 / 77)DLBCL was significantly higher than that in RH group(23.3%,7 / 30),non-GCB and Bcl-2 positive DLBCL The expression level of My D88 was positively correlated with IPI score and Ki-67 proliferation index,and positively correlated with prognosis risk grade and clinical stage.The high expression of My D88 was correlated with poor prognosis of DLBCL patients,but not with gender,age and location of disease.(3)The expression level of mi R-525-5p in DLBCL group was significantly lower than that in RH group.The expression of mi R-525-5p was negatively correlated with clinical stage,My D88,Ki-67 and Bcl-2 expression.The low expression of mi R-525-5p was correlated with poor prognosis of patients,but not with age,gender,location of disease,immune type,IPI score and risk grade.The second part:(1)The high expression of NF-κB / p65 in 35.1%(27 / 77)of DLBCL was related to the expression of non-GCB,My D88 and poor prognosis in DLBCL patients,but notrelated to other clinicopathological parameters.(2)The mutation rates of My D88 l265 P and CD79 B were 5.56%(3 / 54)and 8.33%(1 / 12),respectively.There was no significant correlation with the expression of My D88,NF-κB / p65 protein and the clinical parameters of patients.Conclusions:(1)My D88 stands a good chance of being the target gene of mi R-525-5p;(2)The abnormality of mi R-525-5p and My D88 / NF-κ B pathway is far more likely to be related to the occurrence and development of DLBCL;(3)My D88l265 P and CD79 B mutation rates is too low in systemic DLBCL that except immune immunity sites to be molecular indicators. |
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