The Expression And Clinicopathological Significance Of NSD2,EZH2 And SOX11 In Mantle Cell Lymphoma

Objective Mantle cell lymphoma(MCL)is a kind of clinical heterogeneous disease with high degree of malignancy.the survival time of patients treated with multi-drug combination chemotherapy is still very short,even if patients with new therapeutic drugs and autologous stem cell transplantation still have the risk of recurrence within several years,so it is an incurable disease.Therefore,it is the key to study the mechanism of the occurrence and development of the disease and develop new targeted therapeutic drugs.With the development of sequencing technology and the in-depth study of epigenetics,more attention should be paid to the performance of histone modification in MCL,especially the high-frequency mutations of various genes have been found in the application of sequencing technology.Recently,the mutation of histone methyltransferase NSD2 in MCL was significantly higher than that in foreign countries,and the mutation of EZH2 was also reported.The main purpose of this study was to investigate the expression and mutation of histone methyltransferase NSD2,EZH2 and SOX11 in MCL,and the relationship between the expression level of histone methyltransferase and clinicopathological features,so as to find new targets for the treatment of MCL patients and determine the related clinical predictors for the study of the prognosis of MCL.At the same time,compared with DLBCL,we found the difference in the expression of NSD2 and EZH2 between the two groups,and studied the expression of the two molecules in different diseases.Methods From 2008 to 2019,12 paraffin tissue specimens of MCL were finally diagnosed in the affiliated Hospital of Dali University as the experimental group,and 12 cases of reactive proliferative tonsillitis and It reflects proliferative lymphadenitis were selected as the control group of immunohistochemical(IHC).The expression of NSD2,EZH2 and SOX11 in MCL was detected by IHC,and the expression of NSD2 and EZH2 in DLBCL was studied retrospectively.The data of 76 cases of DLBCL were collected and compared with MCL.The molecular abnormalities were found at the gene level by full exon sequencing in MCL cases.Results In MCL,IHC showed that the expression level of NSD2 was negatively correlated with white blood cell count,NSD2 was positively correlated with Ki67,and NSD2 was positively correlated with LDH.There was a significant difference in the expression of NSD2 between DLBCL and MCL,and there was significant difference between DLBCL and MCL.Mann-Whitney test showed that there was significant difference between the tw o samples,and there was a significant difference between DLBCL and MCL(P<0.05).The expression level of WBC was negatively correlated with leukocyte count(P=0.010<0.05,r=-0.05),and positively correlated with that of NSD2 and LDH(P=0.030<0.05,r=0.707).There was a significant difference between DLBCL and MCL,and there wa s significant difference between the two samples.The expression level of EZH2 was not related to sex,age,ECOG score,clinical stage,white blood cell count,symptom B,KI67,LYM,bone marrow invasion,MIPI score and LDH in MCL patients.There was significant difference in the expression of EZH2 between DLBCL and MCL,and there was significant difference between the two samples by Mann-Whitney test.The expression level of SOX11 was correlated with the sex of MCL patients,but not wit h the age,ECOG score,clinical stage,white blood cell count,B symptom,Ki67,LY M,bone marrow invasion,MIPI and LDH(P>0.05),but the expression level of NSD2 in MCL was related to the expression level of EZH2(P=0.49<0.05).);MCL and D LBCL were not related to the age,ECOG score,clinical stage,white blood cell coun t,B symptom,Ki67,LYM,bone marrow invasion and LDH.There was no statistical significance in the comparison of patient survival curve(P=0.321>0.05).Full exon sequ encing showed that NSD2 was located at 4p16.3.SNP analysis showed that the exon domain of NSD2 produced a mutation of mistranslation,and the difference between b ase and amino acid was NSD2:NM133334:exon1:c.A590G:p.D197G;NSD2:NM133335:e xon2:c.A590G:p.D197G;.NSD2:NM133331:exon3:c.A590 G.p.D197 G,NSD2:NM007331:ex on4:c.A590G:p.D197 G,NSD2:NM133330:exon4:c.A590G:p.D197.There is no In Del exc eption in NSD2.WES showed that EZH2 in MCL was located on 7q36.1 on chromo some 7 SNP found that EZH2 mistranslation mutation occurred in the exon region of MCL,and the protein domain of the mutation site was SANT/Myb.The base and am ino acid of EZH2 were changed to EZH2:NM004456:exon6:c.G553C:p.D185 H.In Del detection showed that frameshift insertion and non-frameshift deletion were found in EZH2 in exon region of MCL patients.EZH2 frameshift insertion bases and amino acids were changed to EZH2:NM004456:exon10:c1139 1140 ins T:p S380 Fs.The non-fra meshift deletion bases and amino acids of EZH2 were changed to EZH2:NM004456:e xon6:c555 563del:p.185 188 del.There were no S0X11-related SNP and In Del abnorm alities in WES sequencing.Sequencing found not only common mutations such as AT M(100%),TP53(50%),CCND1(25%)and KMT2D(50%),but also genes with few mutatio ns in BRCA2(50%),ATRX(50%),CARD11(25%)and NOTCH2(75%).Conclusion In MCL,the expression of NSD2 is positively correlated with that of Ki67,which means that the higher the tumor proliferation,the higher the expression of NSD2.The expression of NSD2 was positively correlated with LDH and negatively correlated with leukocyte count.The expression of NSD2 was different between DLBCL and MCL,but lower in MCL.There was no correlation between the expression of EZH2 and clinical indexes.The expression of EZH2 in DLBCL was significantly different from that in MCL,the expression of EZH2 in MCL was lower than that in DLBCL,and the expression of NSD2 and EZH2 was positively correlated in MCL.The expression of SOX11 is related to the sex of the patients,which is generally high in males and low in females.WES sequencing NSD2 exon region SNP analysis found that there were mistranslation mutations,EZH2 exon region SNP analysis showed existing mistranslation mutations,In Del analysis found frameshift insertion and nonframeshift deletion.