Prognostic value of BCL2 and TP53 genetic alterations for diffuse large B-cell lymphoma patients treated with R-CHOP

Objective:Limited data about the prognostic significance of BCL2 mutations and BCL2 copy number variations in diffuse large B-cell lymphoma(DLBCL)are available.This study aimed to comprehensively describe BCL2 genetic alterations in DLBCL patients,and examine correlation of BCL2,TP53 and other genetic alterations with outcomes in patients treated with R-CHOP.Methods:Probe capture-based high-resolution sequencing was performed on 191 patients diagnosed with de novo DLBCL.MYC,BCL2,and BCL6 protein expressions were detected by immunohistochemistry.Results:The presence of BCL2 alterations significantly correlated with poor progression-free survival(PFS)(5-year PFS:13.7%vs.40.8%;P=0.003)and overall survival(OS)(5-year OS:34.0%vs.70.9%;P=0.036).Importantly,patients who harbored BCL2 gain/amplifications(BCL2GA/AMP)also had a remarkably inferior 5-year PFS(11.1%vs.38.3%;P<0.001)and OS(22.1%vs.69.6%;P=0.009).In contrast,neither BCL2 mutations nor BCL2 translocations were significantly prognostic for survival.Multivariable analyses showed that the presence of BCL2 alterations,especially BCL2GA/AMP,TP53 mutations,and International Prognostic Index(IPI)were significantly associated with inferior PFS and OS.Novel prognostic models for OS were constructed based on 3 risk factors,including BCL2 alterations(Model 1)or BCL2GA/AMP(Model 2),TP53 mutations,and IPI,to stratify patients into 4 risk groups with different survival outcomes.Conclusions:This study showed that DLBCL patients treated with R-CHOP,BCL2 alterations,especially BCL2GA/AMP and TP53 mutations were significantly associated with inferior outcomes,which were independent of the IPI.The novel prognostic models we proposed predicted outcomes for DLBCL patients treated with R-CHOP,but further validation of the prognostic models is still warranted.

Prognostic Significance of BCL2 Protein in Diffuse Large Cell Lymphoma of Head and Neck;Relation to Response to Chemotherapy

Diffuse large B cell lymphoma (DLBCL) is a heterogeneous disease that displays a highly variable clinical outcome. It is a neoplasm of large transformed B cells with a diffuse growth pattern. DLBCL is the most common type of non-Hodgkin’s lymphoma (NHL) (31% of all cases). Approximately half of patients with DLBCL are cured with current chemotherapy regimens. The purpose of this study was to evaluate BCl2 expression in 45 patients diagnosed with DLBCL of head and neck region and correlate the level of its immunohistochemical expression with different clinicopathological variables with emphasis upon patients’ age, gender, nodal or extra-nodal location of lymphoma, patients’ response to chemotherapy, progression-free survival (PFS) and overall survival (OS). A retrospective analysis of 45 patients diagnosed to have DLBCL. A cut off value of ≥ 50% protein expression denoted BCL2 positivity. Out of 45 cases, 36 cases (80%) revealed BCL2 positive expression and 9 cases (20%) were BCL2 negative. We found statistically significant differences in BCL2 expression regarding different patients’ responses to chemotherapy, patients’ OS and PFS (p ≤ 0.05). No statistically significant differences in BCL2 expression regarding the patients’ Ann Arbor clinical stage, age group and tumor site (nodal or extra-nodal, p > 0.05) using the Chi-square test. BCL2 expression was analyzed in relation to 5 years OS and PFS using Kaplan Meier curves and Log Rank test for survival analysis. Cases that demonstrated BCL2 positivity revealed shortened OS and PFS with highly statistically significant differences among the studied variables (p = 0.000). We also found that patients who respond well to the chemotherapeutic regimen had negative BCL2 expression, the differences were statistically significant (p = 0.015). In conclusion, BCL2 expression could be considered a predictor for patients’ chemotherapeutic response, OS and PFS.

Diagnostic and prognostic value of secreted protein acidic and rich in cysteine in the diffuse large B-cell lymphoma

BACKGROUND Secreted protein acidic and rich in cysteine(SPARC)is an extracellular matrixassociated protein.Studies have revealed that SPARC is involved in the cell interaction and function including proliferation,differentiation,and apoptosis.However,the role of SPARC in cancer is controversial,as it was reported as the promoter or suppressor in different cancers.Further,the role of SPARC in lymphoma is unclear.AIM To identify the expression and significance of SPARC in lymphoma,especially in diffuse large B-cell lymphoma(DLBCL).METHODS The expression analysis of SPARC in different cancers was evaluated with Oncomine.The Brune,Eckerle,Piccaluga,Basso,Compagno,Alizadeh,and Rosenwald datasets were included to evaluate the mRNA expression of SPARC in lymphoma.The Cancer Genome Atlas(TCGA)-DLBCL was used to analyze the diagnostic value of SPARC in DLBCL.The Compagno and Brune DLBCL datasets were used for validation.Then,the diagnostic value was evaluated with the receiver operating characteristic(ROC)curve.The Kaplan-Meier plot was conducted with TCGA-DLBCL,and the ROC analysis was performed based on the survival time.Further,the overall survival analysis based on the level of SPARC expression was performed with the GSE4475 and E-TABM-346.The Gene Set Enrichment Analyses(GSEA)was performed to make the underlying mechanism-regulatory networks.RESULTS The pan-cancer analysis of SPARC showed that SPARC was highly expressed in the brain and central nervous system,breast,colon,esophagus,stomach,head and neck,pancreas,and sarcoma,especially in lymphoma.The overexpression of SPARC in lymphoma,especially DLBCL,was confirmed in several datasets.The ROC analysis revealed that SPARC was a valuable diagnostic biomarker.More importantly,compared with DLBCL patients with low SPARC expression,those with higher SPARC expression represented a higher overall survival rate.The ROC analysis showed that SPARC was a favorable prognostic biomarker for DLBCL.Results of the GSEA confirmed that the high expression of SPARC was closely associated with focal adhesion,extracellular matrix receptor interaction,and leukocyte transendothelial migration,which suggested that SPARC may be involved in the regulation of epithelial-mesenchymal transition,KRAS,and myogenesis in DLBCL.CONCLUSION SPARC was highly expressed in DLBCL,and the overexpression of SPARC showed sound diagnostic value.More interestingly,the overexpression of SPARC might be a favorable prognostic biomarker for DLBCL,suggesting that SPARC might be an inducible factor in the development of DLBCL,and inducible SPARC was negative in some oncogenic pathways.All the evidence suggested that inducible SPARC might be a good diagnostic and prognostic biomarker for DLBCL.

CHOP-like Regimen in Combination with Rituximab and Peginterferon Alpha-2b in Newly-diagnosed Diffuse Large B-cell Non-Hodgkin＇s Lymphoma： Experience in a Chinese Center

OBJECTIVE To evaluate the efficacy of rituximab combined with CHOP-like regimen with or without IFN in patients newly diagnosed diffuse large B-cell Non-Hodgkin＇s lymphoma （DLBCL）.METHODS From January 2003 to July 2008, 51 patients received CHOP-like chemotherapy （cyclophosphamide 750 mg/m2, epirubicin 80 mg/m2, vindesine 2.8 mg/m2 on day 1, and prednisolone 100 mg/day on day 1 to day 5）. Thirty-one patients received CHOPR-like treatment （rituximab 375 mg/m2 1 day before CHOP-like chemotherapy）. Twenty patients received CHOP-like regimen in combination with peginterferon （pegIFN） （1μg/kg on day 5） and rituximab （on day 6）.RESULTS -The CR （complete remission） rate in the CHOPR-like （with or without pegIFN） group and in the CHOP-like group was 78.4% and 45.1% （P = 0.005）, respectively. The estimated mean time of overall survival （OS） in the CHOPR-like group and CHOP- like group was 58.7 ± 2.8 and 36.4 ±3.4 months, respectively （P = 0.002）. The rates of CR and OR （overall remission） in CHOPR- like with IFN arm were 85.0% and 95.0%, and the rates of those in CHOPR-like without IFN arm were 74.2% and 87.0% （P 〉 0.05）. The estimated mean time of 4-year-PFS （progression- free survival） in CHOPR-like with IFN arm and in CHOPR-like without IFN arm was 62.9 ±3.0 months and 51.0 ± 4.6 months （P = 0.092）, respectively. In the CHOPR-like with IFN arm, no patient relapsed after achieving CR, while the estimated rate of 4-year- DFS （disease-free survival） in the patients who reached CR in the CHOPR-like without IFN arm was （63.4 ± 19.3）% （P = 0.061）. CONCLUSION Rituximab combined with CHOP-like chemotherapy improved the prognosis of DLBCL patients. The IFN may help to improve the quality and duration of response of DLBCL patients treated with rituximab and CHOP-like regimen.

DETECTION OF BCL-2 GENE MAJOR BREAKPOINT REGION REARRANGEMENT IN HUMAN B-CELL LYMPHOMAS

Objective To investigate the frequency of t(14; 18) in different subtypes of B-cell lymphomas and the ability or the polymerase chain reaction(PCR) to detect this rearrangement in frozen samples. Methods 1o7 cases of B-cell lymphomas were studied uslng DNA extracted from rresh-frozen tissues. The DNA samples were amplified by PCR for bcl-2 MBR/JH. The products of bcl-2/JH rearrangement were hybridized with an internal olignucleotide probe or bcl-2 MBR. Results The rearranged bcl-2MBR/JH gene was detected in 13 of the 25(52. o% ) follicular center lymphomas, according to REAL classification: 8 of 11 (72. 7%) grade 1, 2 of 5(40. 0%) grade I, and 3 of 90 (33. 3%) grade, 17 of 82(2o. 8%) cases or difruse large B-cell lymphomas were found to have detectable bel-2 MBR/J. rearrangement- Conclusion The rrequency or bcl-2 MBR/JH rearrangement in diffuse large B-cell lymphomas is significantly lower than those in follicular center lympkomas(X2= 9. 28, P <o. oo5), suggesting that bcl2/JH rearrangements occur mainly in follicular center lymphomas. in addition, the result of reconstruction experiments suggest that amplification or bcl-2 MBR/JH rearrangements by PCR is both sensitive and specific for detection of t (14; 18 ) translocation.

Effect of rituximab combined with CVAD regimen on serum VEGF and β2-MG levels in patients with primary gastrointestinal B-cell lymphoma

Objective:To investigate the clinical effect of rituximab combined with CVAD regimen in patients with primary gastrointestinal B-cell lymphoma and serum vascular endothelial growth factor (VEGF) andβ2 microglobulin (β2-MG) The impact of the level.Methods:Eighty-four patients with primary gastrointestinal B-cell lymphoma treated from May 2014 to December 2015 were enrolled. Based on the random number table, all the patients were divided into a control group (n=42) and an observation group (n=42). The control group was treated with CVAD. The observation group was treated with rituximab on the basis of the control group. The effect of the patients was evaluated after 3 courses of treatment. The patients were followed up for 3 years after treatment. US RECIST 1.1 was used to evaluate the short-term efficacy on the patients;VEGF, TNF receptor-associated factor 6 (TRAF6) and B-cell lymphoma factor-6 (Bcl-6) levels were measured by enzyme-linked immunosorbent assay;β2-MG level test was implemented to compare the short-term efficacy, biochemical indicators, incidence of toxic side effects and long-term survival rate of the two groups. Results: The short-term efficacy rate of the observation group was 76.19%, which was higher than that of the control group (50.00%) (P<0.05). The levels of VEGF, TRAF6, Bcl-6, andβ2-MG were lower in the observation group after 3 courses of treatment than that in the control group (P<0.05);there was no significant difference in the incidence of neutropenia, gastrointestinal reactions, sepsis, infection, infusion-related reactions and cardiovascular events between the observation group and the control group (P>0.05);The 1-year long-term survival rate after treatment was not statistically significant (P>0.05). The long-term survival rate of the observation group was higher than that of the control group at 2 and 3 years after treatment (P<0.05).Conclusion: The combination of rituximab and CVAD in patients with primary gastrointestinal B-cell lymphoma can improve short-term efficacy, lower VEGF andβ2-MG levels, and lower incidence of side effects. It can improve the long-term survival rate of patients and is worthy of promotion and application.

The Expression and Significance of CyclinD2,MPGES-1,Bcl2 in Diffuse Large B-cell Lymphoma

Objective:To study the expression and significance of cell cycle proteins CyclinD2,mPGES-1,Bcl2 in diffuse large B-cell lymphoma.Methods:Choose lymphoma and sexually hyper-plastic lymphoid tissues as control.Immunohistoc-hemical methods were used to detect the expression of CyclinD2,mPGES-1,and Bcl2,and to compare the positive expression rates of CyclinD2,MPGES-1 and Bcl2 in diffuse large B-cell lymphoma and reactive proliferative lymphoid tissues to compare their diffusion formation.B-cell lymphoma was analyzed for its clinicopathological features.Results:The positive expression rate of CyclinD2,mPGES-1 and Bcl2 in diffuse large B-cell lymphoma is higher than that in reactive proliferative lymphoid tissue,and the difference between the two is statistically significant.There was no statistical difference in CyclinD2,mPGES-1 and Bcl2 in diffuse large B-cell lymphoma between patients according to the age,sex,location,tissue type and degree of differentiation.Conclusion:CyclinD2,mPGES-1 and Bcl2 are highly expressed in patients with diffuse large B-cell lymphoma,and can be used as reference indicators for evaluating the malignant degree and efficacy of dysplasia.

The expression of cyclooxygenase-2 （COX-2） and p16 in non-Hodgkin＇s lymphomas and its clinical significance

Objective： To investigate the expression of cyclooxygenase-2 （COX-2） and p16 proteins in non-Hodgkin＇s lymphomas （NHL） and their relationship with the genesis and progress of it. Methods： The expression of COX-2 and p16 protein were studied in the lymph nodes tissue from 60 NHL patients and 10 control patients with non-malignant diseases by flow cytometry. Results： Positive rate of COX-2 protein expression in NHL tissues （63.3%, 38/60） was higher than that in normal lymphaden tissues （0, 0/10）. The difference was significant between the two groups （P 〈 0.01）. Expression of COX-2 protein was related with the clinical stage of NHL. In stage Ⅰ ＋ Ⅱ patients, it was significantly lower （35.0% ± 54.6%） than that in stage Ⅲ ＋ Ⅳ patients （84.6% ±87.5%） （P 〈 0.01）. In different sex, age, tumor malignant degree, IPI grade, extranodal involvement and B symptoms groups, the differences of COX-2 expression were not statistically significant （P 〉 0.05）. Positive rate of p16 protein expression （41.7%, 25/60） in NHL＇ was statistically lower than that in normal lymphomas （100%, 10/10） （P 〈 0.01）. Expression of p16 protein was related to malignant degree of NHL. The positive rates of p16 protein in low malignant degree tissues （64.7%, 11/17） was higher than that in high malignant degree tissues （14.3%, 2/14） （P 〈 0.05）. Positive rates of p16 protein of NHL tissues in different sex, age, IPI grade, extranodal involvement, clinical stages and B symptoms were not statistically significant （P 〉 0.05）. The p16 protein expression in COX-2 positive patients was 47.4% （18/38）, and in negative patients it was 31.8% （7/22）. There was no statistically difference between them （P 〉 0.05）. Correlation analysis revealed there was no correlation between expression of COX-2 and p16 protein. Conclusion： Both COX-2 and p16 protein may all have relationship with the genesis and progress of NHL. The expression of COX-2 protein in NHL may be a poor prognostic indicator. COX-2 and p16 protein probably have different mechanisms in the genesis and progress of NHL. Their relationship is firstly put forward in this article and needed further studying.

Detection of BCL2-IGHrearrangement on paraffin-embedded tissue sections obtained from a small submucosal tumor of the rectum in a patient with recurrent follicular lymphoma

A 59-year-old woman was admitted to our hospital because of recurrent follicular lymphoma(FL).Colonoscopic examination revealed a rectal submucosal tumor(SMT)without any erosions and ulcers.In this patient,it was difficult to distinguish non-Hodgkin's lymphoma(NHL)invasion from other disorders of the colon including carcinoid tumor merely based on endoscopic findings.Histopathologic and immunohistochemical studies on biopsy specimens showed an infiltration of atypical lymphocytes that were positive for CD20 and BCL2 but negative for UCHL-1.Fluorescence in situ hybridization on paraffin-embedded tissue sections (T-FISH)identified a translocation of BCL2 with IGHgene. Based on these findings,the tumor was defined as an invasion of FL.T-FISH method is useful for the detection of a monoclonality of atypical lymphocytes in an SMT of the gastrointestinal tract,and particularly for the detection of chromosomal translocations specific to lymphoma subtypes.

壁虎多肽混合物对人非小细胞肺癌H1299细胞增殖和凋亡的影响

目的探讨壁虎多肽混合物(GPM)对人非小细胞肺癌H1299细胞增殖和凋亡的影响。方法实验设正常对照组(等体积基础培养液)和GPM高、低剂量组(20,10 mg/mL),采用CCK-8法检测细胞增殖情况,采用流式细胞术检测细胞凋亡情况,采用实时荧光定量聚合酶链反应(RT-qPCR)法和免疫印迹(Western blot)法检测B-细胞淋巴瘤因子2(Bcl-2)、Bcl-2关联X蛋白(Bax)、磷脂酰肌醇-3-激酶(PI3K)mRNA及蛋白表达水平。结果与正常对照组比较,GPM高、低剂量组H1299细胞存活率均显著降低(P<0.01),细胞凋亡率均显著升高(P<0.01),PI3K和Bax mRNA及蛋白表达水平均显著降低(P<0.01),Bcl-2 mRNA及蛋白表达水平均显著升高(P<0.05)。结论GPM能抑制H1299细胞的增殖,其作用机制可能与调控PI3K,Bax,Bcl-2 mRNA及蛋白的表达相关。

晚期胃癌患者参一胶囊联合化疗治疗前后血清Bcl⁃2、Bax、Fas与预后的相关性

目的研究晚期胃癌患者血清B细胞淋巴瘤⁃2基因(Bcl⁃2)、Bcl⁃2相关X蛋白(Bax)、自杀相关因子(Fas)与参一胶囊联合化疗治疗预后的相关性。方法选择2019年1月至2023年4月在安徽省颍上县人民医院进行参一胶囊联合化疗的72例晚期胃癌患者患者并进行1年随访,随访期间存活的患者作为预后良好组(n=48)、死亡的患者作为预后不良组(n=24)。比较两组患者治疗前各项临床资料及血清Bcl⁃2、Bax、Fas水平的差异,采用logistic回归分析预后的影响因素,采用受试者工作特征(ROC)曲线分析血清Bcl⁃2、Bax、Fas对预后的预测效能。结果预后不良组晚期患者参一胶囊联合化疗治疗前的血清癌胚抗原、糖类抗原199、Bcl⁃2水平高于预后良好组,差异有统计学意义(t=4.749、5.032、6.033,P<0.05);血清Bax、Fas水平低于预后良好组,差异有统计学意义(t=5.067、7.018,P<0.05)。治疗前血清Bcl⁃2水平升高是晚期胃癌患者参一胶囊联合化疗治疗预后不良的危险因素,Bax、Fas水平升高是晚期胃癌患者参一胶囊联合化疗治疗预后不良的保护因素(P<0.05)。血清Bcl⁃2、Bax、Fas及血清Bcl⁃2+Bax+Fas均对晚期胃癌患者参一胶囊联合化疗治疗的预后具有预测价值,血清Bcl⁃2+Bax+Fas预测预后的灵敏度和特异度均为87.5%。结论血清Bcl⁃2增加、Bax和Fas降低与晚期胃癌患者参一胶囊联合化疗治疗预后不良有关,治疗前血清Bcl⁃2+Bax+Fas对预后具有预测价值。

Correlation of the abnormal expression of HK-II and TNFAIP3 in diffuse large B-cell lymphoma with the malignant features of tumor cells

Objective: To study the correlation of the abnormal expression of hexokinase-Ⅱ (HK-Ⅱ) and tumor necrosis factor alpha-induced protein 3 (TNFAIP3) in diffuse large B-cell lymphoma (DLBCL) with the malignant features of tumor cells. Methods: DLBCL patients who underwent surgical resection in our hospital between March 2016 and March 2018 were selected as the DLBCL group, and the patients who underwent surgical resection during the same period and were pathologically confirmed as reactive hyperplasia of lymph nodes were selected as the control group. The lesions were collected to measure the expression levels of HK-Ⅱ, TNFAIP3, proliferation genes and invasion genes. Results: HK-Ⅱ mRNA expression level in the lesions of the DLBCL group was significantly higher than that of the control group while TNFAIP3 mRNA expression level was significantly lower than that of the control group;HK-Ⅱ mRNA expression levels in the lesions of DLBCL group of patients with lymphoma stage ⅡI-IV and lymphoma group B were significantly higher than those of the patients with lymphoma stage I-Ⅱ and group A while TNFAIP3 mRNA expression levels were significantly lower than those of the patients with lymphoma stage I-Ⅱ and lymphoma group A;CyclinD2, PDE4B, BCL2, XIAP, CCL5, CXCR4 and MMP26 mRNA expression levels in the lesions of the DLBCL group were significantly higher than those of the control group, positively correlated with HK-Ⅱ and negatively correlated with TNFAIP3 while Caspase-3 and TIMP4 mRNA expression levels were significantly lower than those of the control group, negatively correlated with HK-Ⅱ and positively correlated with TNFAIP3. Conclusion: The high expression of HK-Ⅱ and the low expression of TNFAIP3 in DLBCL are closely related to the pathological process of tumor as well as the proliferation and invasion of tumor cells.

AP2M1抑制弥漫性大B细胞淋巴瘤细胞增殖和侵袭

目的探究接头相关蛋白质复合体2亚基μ1(AP2M1)对弥漫性大B细胞淋巴瘤(DLBCL)细胞增殖和侵袭的调控作用。方法将人弥漫性大B细胞淋巴瘤细胞系OCI-LY8分为对照组、NC-LV组、AP2M1-LV组。用Lipofectamine 2000进行细胞转染。四甲基偶氮唑盐(MTT)法检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell小室法检测细胞迁移和侵袭。Western blot检测AP2M1、表皮生长因子受体(EGFR)、p-磷脂酰肌醇3激酶(PI3K)和p-蛋白质激酶B(AKT)蛋白表达。结果与对照组相比,AP2M1-shRNA组细胞中AP2M1的mRNA和蛋白相对表达量均降低(P<0.05),相对细胞活力升高(P<0.05),细胞凋亡率降低(P<0.05),迁移和侵袭细胞数量均升高(P<0.05);EGFR的蛋白相对表达量及PI3K和AKT的磷酸化水平均升高(P<0.05)。与对照组相比,AP2M1-LV组细胞中AP2M1的mRNA和蛋白相对表达量均升高(P<0.05),相对细胞活力降低(P<0.05),细胞凋亡率升高(P<0.05),迁移和侵袭细胞数量均降低(P<0.05),EGFR的蛋白相对表达量及PI3K和AKT的磷酸化水平均降低(P<0.05)。结论AP2M1的过表达部分通过抑制EGFR/PI3K/AKT信号通路来抑制DLBCL细胞的增殖和侵袭。

湿润烧伤膏对糖尿病足创面组织中MMP-2、MMP-9、Bcl-2、Bax水平的影响

目的探讨湿润烧伤膏(MEBO)对糖尿病足创面组织中基质金属蛋白酶(MMP)-2、MMP-9、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)水平的影响。方法选取2020年5月至2022年5月郑州大学附属郑州中心医院收治的55例糖尿病足患者作为研究对象,按照不同治疗方法将其分为MEBO组(35例)和VSD组(20例),MEBO组患者局部创面采用MEBO治疗,VSD组患者局部创面采用负压封闭引流(VSD)治疗,对比观察两组患者创面面积,创面组织中MMP-2、MMP-9、Bcl-2、Bax水平及临床疗效。结果治疗第7、21天,MEBO组患者创面面积均明显小于VSD组(t=2.719、5.268,P=0.009、P<0.001),创面组织中MMP-2、MMP-9、Bax水平均明显低于VSD组(MMP-2:t=2.138、2.202,P=0.037、0.032;MMP-9:t=2.129、2.476,P=0.038、0.017;Bax:t=3.623、3.038,P=0.001、0.004),Bcl-2水平均明显高于VSD组(t=2.040、3.054,P=0.046、0.004);治疗21 d后,MEBO组患者中显效23例、有效10例、无效2例,明显优于VSD组患者的显效8例、有效7例、无效5例(Z=-2.126,P=0.033)。结论MEBO可通过降低糖尿病足创面组织中MMP-2、MMP-9、Bax水平以及提高Bcl-2水平促进创面愈合。

MYC蛋白与BCL-2蛋白双表达DLBCL患者临床特点与预后影响因素分析

目的分析细胞致瘤基因(cellular-myelocytomatosis viral oncogene,MYC)蛋白与B细胞淋巴瘤基因-2(B cell lymphoma/lewkmia-2,BCL-2)蛋白双表达弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)患者的临床特点与预后影响因素。方法选取2020年1月—2021年1月南京医科大学附属苏州医院收治的82例DLBCL患者,均行免疫组化检测MYC蛋白与BCL-2蛋白表达情况,并连续随访3年,统计患者的生存时间,分析预后影响因素。结果MYC/BCL-2蛋白双表达组DLBCL患者改良AnnArbor分期Ⅲ~Ⅳ期、肿瘤包块直径≥5 cm、淋巴瘤国际预后指数(international prognostic score,IPS)评分3~4分及骨髓侵犯占比均高于非MYC/BCL-2蛋白双表达组,差异有统计学意义(P<0.05)。MYC/BCL-2蛋白双表达组DLBCL患者平均生存时间为(22.35±2.48)个月,非MYC/BCL-2蛋白双表达组平均生存时间为(30.70±1.66)个月,MYC/BCL-2蛋白双表达组DLBCL患者平均生存时间低于非MYC/BCL-2蛋白双表达组,差异有统计学意义(P<0.001)。logistic回归分析显示,改良Ann-Arbor分期Ⅲ~Ⅳ期、骨髓侵犯、MYC/BCL-2蛋白双表达为影响DLBCL患者预后的危险因素(P<0.05)。结论MYC蛋白与BCL-2蛋白双表达DLBCL患者临床特征主要表现为分期晚、肿瘤大包块、更易发生骨髓侵犯且预后差;MYC蛋白与BCL-2蛋白双表达可作为DLBCL患者预后的影响因素。文章中体现了《中国弥漫大B细胞淋巴瘤诊断与治疗指南(2013年版)》的临床参考或执行标准。

Lenalidomide regulates the CCL21/CCR7/ERK1/2 axis to inhibit migration and proliferation in diffuse large B-cell lymphoma

Background:The prognostic significance of the chemokine receptor CCR7 in diffuse large B-cell lymphoma(DLBCL)has been reported previously.However,the detailed mechanisms of CCR7 in DLBCL,particularly regarding its interaction with lenalidomide treatment,are not fully understood.Methods:Our study utilized bioinformatics approaches to identify hub genes in SU-DHL-2 cell lines treated with lenalidomide compared to control groups.Immunohistochemical data and clinical information from 122 patients with DLBCL were analyzed to assess the correlation of CCR7 and p-ERK1/2 expression with the prognosis of DLBCL.Furthermore,in vitro and in vivo experiments were conducted to clarify the role of CCR7 in the response of DLBCL to lenalidomide treatment.Results:Our bioinformatics analysis pinpointed CCR7 as a hub gene in the context of lenalidomide treatment in DLBCL.Notably,31.14%and 36.0%(44/122)of DLBCL cases showed positive expression for CCR7 and ERK1/2 respectively,establishing them as independent prognostic factors for adverse outcomes in DLBCL via multivariate Cox regression analysis.Additionally,our studies demonstrated that the external application of the protein CCL21 promoted proliferation,migration,invasion,and activation of the ERK1/2 pathway in SU-DHL-2 and OCI-LY3 cell lines with high levels of CCR7 expression.This effect was mitigated by CCR7 silencing through siRNA,application of ERK inhibitors,or lenalidomide treatment.In vivo experiments reinforced the efficacy of lenalidomide,significantly reducing tumor growth rate,tumor mass,serum total LDH levels,and expression of CCR7 and p-ERK1/2 in a SUDHL-2 xenograft model in nude mice(p<0.05).Conclusion:Our study clarifies the potential role of the CCL21/CCR7/ERK1/2 axis in the therapeutic effects of lenalidomide in DLBCL treatment.

BCL-2蛋白在弥漫大B细胞淋巴瘤中的表达及预后价值

目的:探讨BCL-2蛋白表达在美罗华时代弥漫大B细胞淋巴瘤(DLBCL)中的预后价值。方法:对我院自2008年1月至2010年12月128例初诊为DLBCL的患者临床资料进行回顾性分析,比较不同BCL-2蛋白表达组中DLBCL的预后。结果:83例(64.8%)患者BCL-2蛋白表达阳性,45例(35.2%)患者BCL-2蛋白表达阴性。全部DLBCL病人中,BCL-2蛋白表达不能作为判断DLBCL预后的独立因素(3年OS:76.6%vs 76.8%,P=0.960;3年PFS:57.1%vs 70.5%,P=0.344);在60岁及以上DLBCL患者中,BCL-2蛋白阳性组与BCL-2蛋白阴性组相比3年总生存率无显著差异(66.7%vs 76.4%,P=0.133),但是3年无进展生存率明显低于BCL-2蛋白阴性组(35.8%vs 83.3%,P=0.037)。结论:肿瘤细胞BCL-2蛋白表达阳性是60岁以上DLBCL患者的不良预后因素,在该组患者中具有明确的预后价值。

甘石青黛膏对大鼠亚急性湿疹模型皮损淋巴细胞和角质形成细胞Bcl-2与Bax蛋白表达变化的影响

目的观察甘石青黛膏对大鼠亚急性湿疹模型皮损淋巴细胞和角质形成细胞B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白与Bax蛋白表达变化的影响,探究中药外治湿疹的分子生物学依据。方法造模前取大鼠正常皮肤组织,制作成石蜡切片备用,造模成功后分为治疗组和模型组,治疗组采用中药甘石青黛膏干预,模型组无干预,检测三组大鼠Bcl-2与Bax蛋白表达情况。结果造模前大鼠Bcl-2与Bax蛋白表达均呈阳性,且Bcl-2/Bax比率接近于1;治疗组Bcl-2蛋白表达呈弱阳性,模型组Bcl-2蛋白表达呈强阳性,两者相比具有显著统计学差异(P<0.001);治疗组Bax蛋白表达呈阳性,模型组Bax蛋白表达呈弱阳性,两者相比具有显著统计学差异(P<0.001);治疗组与模型组Bcl-2/Bax比率相比具有显著统计学差异(P<0.001),后者明显高于前者;治疗组与健康组Bcl-2/Bax比率相比无统计学差(P>0.05)。结论甘石青黛膏具有抑制Bcl-2蛋白表达,促进Bax蛋白表达,使细胞凋亡调控系统趋于平衡的作用。

高血脂大鼠脑缺血再灌注损伤后Bcl-2和Bax的表达变化及灯盏乙素的影响

目的观察高血脂大鼠缺血侧大脑皮质额叶内B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达变化,探讨灯盏乙素对高血脂大鼠脑缺血再灌注损伤的保护作用。方法大鼠建立高血脂模型后,线栓法阻塞大脑中动脉建立脑缺血再灌注损伤模型,HE染色观察大脑皮质额叶的病理变化,免疫组织化学和免疫印迹法观察Bcl-2、Bax蛋白的表达变化,同时结合神经行为学评分和2,3,5-氯化三苯基四氮唑(TTC)染色观察大鼠的神经行为及脑梗死灶体积的变化。结果与假手术组相比,生理盐水组大鼠脑组织损伤加重,Bcl-2的表达明显减少,Bax的表达明显增高,同时大鼠的神经行为学评分和脑组织梗死灶体积明显增加(P<0. 05)。与生理盐水组相比,灯盏乙素治疗组脑组织损伤减轻,Bcl-2的表达明显增多,Bax的表达明显减少,同时大鼠的神经行为学评分和脑组织梗死灶体积明显降低(P<0. 05)。结论灯盏乙素对高血脂大鼠脑缺血再灌注损伤的保护作用可能是通过促进Bcl-2的表达,抑制Bax的表达实现的。

针刺介入时机对缺血性中风大鼠神经功能评分和Bcl-2、Bax表达的影响

目的研究不同时间点介入针刺治疗对缺血性中风大鼠神经功能评分和对缺血脑组织中B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)阳性表达的细胞数量的影响,探讨针刺治疗机制及介入时机。方法将200只雄性SD大鼠随机分为假手术组、模型组、2小时针刺组、72小时针刺组和168小时针刺组5组,除假手术组外,均采用线栓法制备大鼠局灶性永久性脑缺血模型,在术后1、3、7、14天,4个时间点采用Garcia复合评分法评价大鼠神经功能,用酶联免疫分析法探查缺血脑组织中的Bcl-2、Bax蛋白表达情况,并在显微镜下记录阳性细胞数目进行比较。结果针刺各组缺血性中风大鼠神经功能评分得到明显改善,优于模型组(P<0.001);2小时针刺组分别优于72小时、168小时组(P<0.01),差异有统计学意义。缺血性中风大鼠术后不同时间点脑组织中的Bcl-2,Bax表达均增高,1天达到高峰,随后下降,与假手术组比较有统计学意义(P<0.001,P<0.01)。术后2小时与模型组、72小时、168小时针刺组比较,可明显提高Bcl-2蛋白的表达、抑制Bax蛋白的表达,差异有统计学意义(P<0.001);72小时与168小时针刺组比较可明显提高Bcl-2蛋白的表达、抑制Bax蛋白的表达,差异有统计学意义(P<0.01)。结论针刺可明显提高缺血性中风大鼠的Garcia评分,改善其运动、感觉和协调功能;针刺可通过调节Bcl-2、Bax蛋白表达来抑制神经细胞凋亡,挽救缺血半暗带神经细胞的功能,而且在缺血性中风后越早介入针刺治疗疗效越好,存在一个最佳治疗时间窗。