| Objective: To analyze the protein expression and upstream mi RNA of EZH2 and NSD2 in diffuse large B-cell lymphoma(DLBCL),and to explore the relationship between the role of EZH2 and NSD2 in the occurrence and development of DLBCL and clinical pathological characteristics and prognosis.Methods: Collect 108 specimens of the first diagnosis of DLBCL in the First Affiliated Hospital of Dali University from 2008 to 2018 and 20 specimens of reactive hyperplasia of lymph nodes(RH)during the same period.Part 1:(1)Immunohistochemistry(IHC)En Vision two-step method was used to detect the expression of EZH2 and NSD2 in DLBCL and RH.(2)Through SPearman correlation analysis and chi-square test,analyze the relationship between EZH2 and NSD2 protein expression in DLBCL and the pathological characteristics of patients.(3)Analyze the relationship between EZH2 and NSD2 and the prognosis of patients by Kaplan-Meier.Part 2:(1)Using mi RNA chip technology to detect mi RNA in 10 cases of DLBCL and 8 cases of RH,and analyze the difference mi RNA.(2)Use targetscan and mi Rtargets two databases to predict and analyze the upstream mi RNA changes that regulate EZH2 and NSD2 in differential mi RNAs.Results: Part 1:(1)The protein expression rates of EZH2 and NSD2 in DLBCL were73.8% and 61.8%,respectively.The positive signal of RHL in the control group only appeared in the germinal center.(2)The expression of EZH2 and NSD2 in DLBCL has a positive correlation(rs=0.214,P=0.031,P<0.05).(3)The expression of EZH2 protein is related to the site of DLBCL patients(P=0.013,P<0.05),the expression outside the nodules is higher than that inside the nodules(P=0.013,P<0.05);the expression of EZH2 protein is related to the expression of C-MYC Correlation(P=0.014,P<0.05),EZH2 is more likely to be highly expressed in the C-MYC positive group(P=0.014,P<0.05);EZH2 is correlated with the expression of BCL-6(P=0.014,P <0.05),EZH2 is more likely to be highly expressed in the BCL-6 positive group(P=0.015,P<0.05);EZH2 and BCL-2 expression have a relative tendency(P=0.06,P<0.05),EZH2 is more inclined High expression in BCL-2 positive group(P=0.059,P≈0.05);EZH2 protein expression and patient gender(P=0.086),age of onset(P=0.744),immunophenotyping(P=0.0.906),clinical Staging(P=0.376),has nothing to do with IPI score(P=0.571),CD5(P=0.459),CD30(P=0.655)and Ki-67(P=0.858)expression.(4)NSD2 protein expression has a tendency to be correlated with C-MYC expression(P=0.063,P≈0.05),NSD2 is more likely to be highly expressed in the C-MYC positive group;NSD2 protein expression is not related to age of onset(P=0.363),gender(P=0.119),immunophenotyping(P=0.848),clinical stage(P=0.44),irrelevant site(P=0.371),IPI score(P=0.620),CD5(P=0.459),CD30(P =0.655)and Ki-67(P=0.858).(5)The expression of EZH2 and NSD2 protein was not related to the overall survival time of DLBCL patients(PEZH2=0.821,PNSD2=0.768,PEZH2-NSD2=0.456,P value>0.05).Part 2:(1)There were126 differential mi RNAs in the detection of mi RNA between the DLBCL and the control group RHL,of which 59 mi RNAs were up-regulated and 67 mi RNAs were expressed.(2)The levels of hsa-mi R-26a-5p that may be related to the regulation of EZH2 in differential mi RNAs were significantly reduced.(3)Among the mi RNAs that may be related to the regulation of NSD2 in differential mi RNAs,the expression levels of which have decreased are hsa-mi R-875-3p,hsa-mi R-622,hsa-mi R-5003-3p,hsa-mi R-497-5p,hsa-mi R-4676-3p,hsa-mi R-4328,hsa-mi R-3934-5p,hsa-mi R-3918,hsa-mi R-26a-5p,hsa-mi R-200c-3p;mi RNA expression level is up-regulated by hsa-mi R-5004-5p,hsa-mi R-4694-3p,hsa-mi R-4490,hsa-mi R-4284,hsa-mi R-4264.Conclusion: EZH2 and NSD2 proteins have a synergistic effect in the occurrence and development of DLBCL.EZH2,NSD2 and related upstream mi RNAs may participate in the development and development of DLBCL. |
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