A Study Of The Relationship Of SOX11mRNA,miR-155-5p,miR-34a And P53 Gene Variation,Clinical Pathology Characteristics And Prognostic Significance In Mantle Cell Lymphoma

Objective:Mantle cell lymphoma(MCL)is a subgroup of aggressive B cell non-hodgkin's lymphoma(NHL),which has poor prognosis and the median survival was only 3 to 5years.Meanwhile MCL is a heterogeneous disease,not all cases present aggressive course,10-15%cases had indolent course.So it is necessary to explore new molecular genetic markers and prognostic factors in MCL to provide basis for treatment strategies.The specific hallmark in MCL is t(11;14)that led to the high expression of CCND1.The mantle cell lymphoma international prognostic index(MIPI)was used as a prognostic factor in stratified treatment.P53 gene variation was associated with the poor prognosis of patients with MCL including gene mutations and gene deletion.Studies have shown that P53 gene mutation was correlation with P53 protein strong expression,as a result protein expression could be detected when detecting gene mutation was difficult.SOX11 has important diagnostic value in MCL,whether it has prognostic value in MCL was unconclued.Some studies have shown that high SOX11 expression is associated with good prognosis,on the contrary,there are also some opposite conclusion.A variety of microRNA regulating the expression of target genes play an important role in pathogenesis and might be related to prognosis in lymphoma.Abnormal expression of miR-17-92,miR-155,miR-150 and miR-34 is a common event in the process of B cell tumor.miR-34a involved in the regulation of network surrounding P53 gene through FOXP1 and BCL2 affect the B cell differentiation and apoptosis.Low expression of miR-34a in diffuse large B lymphoma(BLBCL)is associated with prognosis.MiR-155 is the first micro RNA confirmed that could induce lymphoma.High expression of miR-155 is related to the transformation from monoclonal B lymphocyte(MBL)to CLL,poor response to treatment in CLL and short survival.This study examining expression of SOX11mRNA,miR-34a and miR-155-5p and P53 gene variation in 75 patients with MCL explore the relationship of SOX11mRNA,miR-34a,mi R-155-5p and P53 gene variation,pathological characteristics such as staging,immune phenotype,MIPI and prognostic significance in mantle cell lymphoma.Methods:1.The detailed clinical data of 75 patients with MCL were collected.All cases were followed up by telephone,and the median follow-up time was 40(2-98)months.2.The mantle cell lymphoma international prognostic index(MIPI)were rechecked in all cases according to simplified(s MIPI)formula.3.Tissue microarray was prepared using tissue array instrument.4.Immunohistochemical methods were used to detect the expression of CCND1,Ki67,SOX11 and P53 in 75 patients with MCL.5.Fluorescence in situ hybridization was used to detect P53 gene deletion in 75patients with MCL..6.exon5-9 mutation of P53 gene was detected by DNA sequencing in 25 patients with MCL.7.Fluorescence quantitative PCR method was used to detect the expression level of SOX11mRNA in 75 patients with MCL..8.Taking advantage of bioinformation database TargetScan and mi Rdb predicted miRNA that might regulate expression of SOX11.9.Fluorescence quantitative PCR method was used to detect the expression levels of miR-155-5p and miR-34a in 75 patients with MCL.10.A variety of statistical methods were used to analyze the expression of SOX11mRNA in MCL patients,the correlation between SOX11mRNA and clinicopathological features and P53 gene variation,prognostic value of SOX11mRNA expression and P53 gene variation in patients with MCL.Statistical methods were used to investigate expression of miR-155-5p and miR-34a in patients with MCL,the correlation between miR-155-5p and clinicopathological features and SOX11m RNA,the correlation between miR-34a and clinicopathological features and P53 expression,and prognostic significance of miR-155-5p and miR-34a in patients with MCL.Results:1.Clinical characteristicsAmong 75 cases of MCL,58 cases were male(77%),17 cases were female(23%),and age was 39 to 79 years,median age was 63 years.48 cases(64%)were ECOG2-4points,and 56 cases(75%)were Ann Arbor stage?~?.there were B symptoms in 13cases(17%),22 cases(29%)involved in bone marrow.Peripheral blood cells were more than 10×10~9/L in 11 cases(15%),and LDH elevated in 17 cases(23%),and?-2microglobulin(?2-MG)increased in 35 cases(47%).21(28%)cases,31(41%)cases and23(31%)cases were respectively in low-risk group,intermediate–risk group and high-risk group according to MIPI.2.Histopathological and immunological characteristics69 cases(92%)were in classical histomorphology and 6 cases(8%)present blastiod transformation.Positive expression in immunohistochemistry were 100%(75/75)in cyclinD1,97%(66/68)in bcl-2,82%(60/73)in CD5,21%(15/72)in CD23,12%(8/69)in bcl-6,and 29%(22/75)in Ki67?30%,respectively.SOX11 positive were in 97%(73/75)cases and P53 positive in 21%(16/75)cases.3.P53 gene aberration were in 16/75(26%)cases.4.Expression of SOX11mRNA were detected by RT-PCR.The result revealed SOX11mRNA level 3.097(1.311,6.216)in patients with MCL,and control group 1.058(0.302,2.623).The expression level of SOX11mRNA in patients with MCL was significantly higher than that in reactive hyperplasia lymphoid(p=0.00015).5.Correlation between SOX11mRNA expression and clinicopathological features and P53 gene aberration.5.1 Correlation between SOX11mRNA expression and clinicopathological features.Expression of SOX11mRNA in cases with ECOG<2,normal bone marrow were respectively higher than that in cases with ECOG?2(P=0.0011),bone marrow involved(P=0.0038).It was statistically different between MIPI three groups(P=0.0031),furthermore being analyzed by the multiple comparison,and it was statistically different betweengrouplow-riskandgrouphigh-risk,grouplow-riskandgroup intermediate-risk,respectively,however,it wasn't statistical different between group intermediate-risk and group high-risk.5.2 Correlation between SOX11mRNA expression and P53 gene aberration.SOX11mRNA expression was3.742(2.263,5.717)in P53 wide type group,3.778(1.386,6.921)in P53 aberration group,and there was no statistical difference between the two groups in SOX11mRNA expression(P=0.6662).6.Survival analysis of SOX11mRNA expression,P53 gene aberration and MIPI in MCL patients.6.1 Survival analysis of SOX11mRNA expression in patients with MCL.Median survival in 75 cases with MCL was 33(30-38)months.Cases were divided into group SOX11mRNA?M and group SOX11mRNA<M according to SOX11mRNA level.Median survival is shorter in SOX11mRNA<M group than that in SOX11mRNA?M group,(27 months&50 months).There was statistically significant differences between the two groups(P<0.001).6.2 Survival analysis of P53 gene aberration in patients with MCLMedian survival time was 30 months in TP53 aberration group,36 months in TP53 wide type group,and difference between two groups was statistically significa-nt(P<0.05).Furthermore,cases were divided into 4 groups according to TP53 m utation or/and deletion:TP53mut+,TP53del+,TP53mut+del+,TP53mut-del-.Median surv ival time was 30?25?31.5?37 months,respectively(P<0.05).6.3 Survival analysis of TP53 gene aberration combined SOX11mRNA expression in patients with MCLFurthermore,cases were divided into 4 groups according to the level SOX11m RNA expression and TP53 aberration:TP53abe SOX11mRNA?M;P53+SOX11m RN A<M;P53-SOX11mRNA?M;P53-SOX11mRNA<M;,there was significant statistical difference between-the survival curves of the 4 groups(P<.0001).The best surviva l was in P53+SO-X11m RNA?M,and the worst survival was P53+SOX11m RNA<M group.6.4 Survival analysis of MIPI combined SOX11mRNA expression in patients with MCLMedian survival was 44 months in cases with low-risk,31 months in cases with intermediate-risk,30 months in cases with high-risk.There was statistically difference between low-risk and high-risk group(p=0.0033)and low-risk group and intermediate-risk group,however,no statistical difference between the intermediate-risk and high-risk group.Furthermore,cases were divided into group SOX11mRNA<M and group SOX11mRNA?M in every MIPI group according to SOX11mRNA expression,and median survival time were shorter in group SOX11mRNA<M than that in SOX11mRNA?M(p<0.05).7.Expression of miR-34a and miR-155-5p were detected by RT-PCR.Expression of miR-34a was 0.478(0.202,0.821)in cases with MCL and 0.572(0.265,1.072)in the control group.There was no significant difference in expression of miR-34a between the two groups(P=0.335).Expression of miR-155-5p was 3.482(2.136,5.048)in cases with MCL and 0.774(0.083,1.834)in the control group.Expression of miR-155-5p in cases with MCL was significantly higher than that in patients with RHF(P<0.001).8.Correlation between miR-34a and miR-155-5p expression and clinicopathological features.8.1 Correlation between mir-34a expression and clinicopathological features.Expression of miR-34a was 0.385(0.184,0.69)in cases with ECOG?2,0.768(0.228,0.976)in cases with ECOG<2,and it was significantly lower in group ECOG?than that in group ECOG<2(P=0.0214).The expression of miR-34a was0.162(0.107,0.362)in cases with SOX11m RNA<M,0.503(0.219,0.886)in cases with SOX11mRNA?M,and it was significantly lower in group SOX11mRNA<M than that in group SOX11mRNA?M(P=0.0309).8.2 Correlation between miR-155-5p expression and clinicopathological featuresExpression of miR-155-5p was 3.031(1.27,1.27)in cases with low-risk,4.438(2.99,2.99)in cases with intermediate-risk,4.438(2.704,2.704)in cases with high-risk group.It was statistically different between the three groups(P=0.0432),furthermore being analyzed by multiple comparison,and it was significantly lower in group low-risk than that in group intermediate-risk and group high-risk,respectively,howerve,it wasn't statistically different between group intermediate-risk and group high-risk.9.Survival analysis of expression of miR-34a and miR-155-5p in MCL patients9.1 survival analysis of mi R-34a expression in patients with MCL.Cases were divided into group miR-34a?M and group miR-34a<M according to expression level of miR-34a.Median survival was 39 months in group miR-34a?M,28months in group miR-34a<M,and it was statistically significant between the two groups(p=0.0049).9.2 survival analysis of mi R-155-5p expression in MCL patients.Cases were divided into group miR-155-5p?M and group miR-155-5p<M according to expression level of miR-155-5p.Median survival was 31 months in group miR-155-5p?M,36 months in group miR-155-5p<M,and it was statistically significant between the two groups(p=0.0281).10.Prognostic analysis of SOX11mRNA expression in patients with MCLAt alpha=0.05 level,univariate analysis showed poorer survival was associated with blastiod transformation,ECOG?2,P53+,bone marrow involvement,high-risk group and SOX11mRNA<M,miR-34a<M and miR-155-5p?M,however,multivariate analysis showed only blastiod transformation,high-risk group and SOX11mRNA<M were poor prognostic factors.Conclusion:1.Expression of SOX11mRNA in patients with MCL is significantly higher than that in control group.Compared with SOX11m RNA<M,SOX11mRNA?M might be good prognostic factor,be associated with longer Survival and might be more accurate to predict prognosis combined with MIPI or P53 expression.2.Although mi R-34a<M and miR-155-5p?M were excluded out of prognostic model by multivariate analysis,univariate analysis showed median survival in group miR-34a<M and group miR-155-5p?M was significantly shorter than that in group miR-34a?M and miR-155-5p<M,respectively.Expression of miR-34a and miR-155-5p might be have potential prognostic value in MCL,more studies need to be performed in the future.