【作者】 王艳；

【导师】 吴慧；

【作者基本信息】 郑州大学 ， 肿瘤学（专业学位）， 2022， 硕士

【摘要】 第一部分B7-H3在IDH野生型GBM中的表达及其与预后的关系目的1.分析B7-H3在IDH野生型胶质母细胞瘤(glioblastoma,GBM)组织中的表达及其与放化疗敏感性的关系。2.联合分析B7-H3和MGMT对IDH野生型GBM预后的影响。方法1.回顾性分析2015年1月至2019年1月就诊于郑州大学附属肿瘤医院的65例IDH野生型GBM患者的一般资料及病理资料。所有患者均行标准的Stupp方案治疗,包括手术治疗、术后口服替莫唑胺(temozolomide,TMZ)行同期放化疗和辅助化疗。2.免疫组化法用于检测B7-H3在GBM肿瘤组织中的表达水平,Image-ProPlus(6.0)图像分析软件用于定量测量B7-H3的表达强度,以平均光密度(mean optical density,MOD)表示强度。3.获取并分析65例GBM患者接受同期放化疗前、后的磁共振影像学资料。3D-Slicer(4.11.20210226)可视化画软件用于测量患者接受同期放化疗前后肿瘤体积变化,以评价近期疗效。根据Mac Donald评价标准,患者被分为完全缓解(complete response,CR),部分缓解(partial Remission,PR),病情稳定(stable Disease,SD)和疾病进展(progressive Disease,PD)组。并定义敏感组:CR+PR,抗拒组:PD+SD。4.受试者工作特征曲线(receiver operator curve,ROC)用于评价B7-H3表达强度对IDH野生型GBM放化疗的敏感性的诊断价值及确定cut-off值。5.根据B7-H3在肿瘤组织中的表达水平,患者被分为高表达和低表达组,联合分析B7-H3表达水平和MGMT甲基化状态对GBM患者预后的影响。6.统计学分析:SPSS(26.0)版统计学软件用于数据分析。卡方检验和Fisher精确检验用于比较患者的临床基本特征,采用Kaplan–Meier法计算OS和PFS,并用Log-Rank法检验。P<0.05时认为有统计学意义。7.随访:患者于同期放化疗结束后1个月复查头颅MRI,此后,每3个月复查一次头颅MRI。随访时间截至2020年12月。结果1.Image-Pro Plus软件分析结果显示,65例IDH野生型GBM患者B7-H3的MOD值范围为0.0347～0.2355,中位MOD值为0.1262。B7-H3低表达35例,高表达30例。2.近期疗效:患者CR:19例,PR:28例,SD:10例,PD:8例。敏感组47例,抗拒组18例。t检验分析结果显示:放射抗拒组的MOD值明显高于放射性敏感组(0.1648±0.0426 vs.0.1164±0.439),P=0.001,差异有统计学意义。3.ROC曲线分析结果显示,肿瘤组织中B7-H3的表达强度对GBM放化疗敏感性有诊断价值。AUC(area under curve)=0.778;P=0.001;95CI为0.670～0.926;cut-off值为0.1471,敏感性为75.0%,特异性为76.6%。4.(1)B7-H3高表达组患者30例,其中MGMT甲基化14例,MGMT未甲基化16例。MGMT甲基化组和MGMT未甲基化组中位PFS分别为8个月、7个月,P>0.05,两组无统计学差异。MGMT甲基化组与未甲基化的中位OS为分别为15个月、14个月,P>0.05,两组无统计学差异。(2)B7-H3低表达组患者35例,其中MGMT甲基化19例,MGMT未甲基化16例。MGMT甲基化组与未甲基化组中位PFS分别为20个月、10个月,P<0.05,两组有统计学差异。MGMT甲基化组与未甲基化组中位OS分别为30个月、18个月,P<0.05,两组有统计学差异。结论B7-H3的表达水平影响IDH-野生型GBM放化疗敏感性,B7-H3低表达的GBM患者MGMT启动子甲基化患者预后更好。B7-H3可能成为GBM预后的新的分子标志物。第二部分分析CDKN2A的表达和MGMT甲基化状态对IDH野生型GBM预后的影响目的1.分析65例IDH野生型GBM患者细胞周期蛋白依赖性激酶抑制蛋白2A(cyclin-dependent kinase inhibitor 2A,CDKN2A)的表达状态;2.联合分析CDKN2A基因状态与MGMT甲基化状态对IDH-野生型GBM预后的影响。方法1.采用荧光原位杂交(fluorescent in situ hybridazation,FISH)检测技术分析65例IDH野生型GBM肿瘤组织中CDKN2A基因状态,根据检测结果将患者分为CDKN2A纯合性缺失型和CDKN2A表达型。2.Kaplan-meier法用于计算生存时间,并用Log-rank法检验,P值<0.05有统计学意义。结果1.FISH检测结果显示,65例GBM患者中26例为CDKN2A纯合性缺失型,39例为CDKN2A表达型。2.生存分析(1)CDKN2A缺失组CDKN2A纯合性缺失组26例,MGMT甲基化患者11例,MGMT未甲基化患者15例。MGMT未甲基化的中位PFS为7个月,MGMT甲基化中位PFS为8个月,差异无统计学意义(P=0.076);MGMT未甲基化的中位OS为10个月,MGMT甲基化中位OS为13个月,差异无统计学意义(P=0.201)。(2)CDKN2A表达组CDKN2A未缺失患者39例,其中MGMT甲基化22例,MGMT未甲基化17例。MGMT未甲基化的中位PFS为12个月,MGMT甲基化中位PFS为26个月,差异有统计学意义(P=0.006);MGMT未甲基化的中位OS为18个月,MGMT甲基化中位OS为29个月,差有统计学意义(P=0.0027)。结论MGMT甲基化状态对IDH野生型GBM患者预后的影响与CDKN2A基因状态显著相关。更多还原

【Abstract】 Section 1 Effect of B7-H3 expression on prognosis of IDH-wild type GBM Objectives1.To explore the effect of B7-H3 expression on the chemoradiosensitivity of patients with IDH-wild type glioblastoma(GBM).2.To investigate the effect of B7-H3 expression level combined with MGMT promoter methylation status on prognosis of IDH-wild type GBM.Method1.We conducted a retrospective review of the clinical and pathologic characteristics of 65 adult patients with IDH‐wild type GBM who were treated in Affiliated Cancer Hospital of Zhengzhou University from January 2015 to December2019.All patients accepted Stupp’s regimen as standard treatment,which involved surgical resection followed by concurrent radiotherapy and adjuvant chemotherapy based on temozolomide(TMZ).2.Immunohistochemistry(IHC)was used to detect B7-H3 expression level in glioma tissue.Image-Pro-Plus(6.0)was applied to analysis of the fluorescence intensity,which indicated with mean optical density(MOD).3.Pre-and post-concurrent chemoradiotherapy MRI data of 65 patients were retrospectively analyzed.3D Slicer visualization software(4.11.20210226)was used to access tumor volumes and evaluate the short-term outcomes.Patients were categorized into complete remission(CR),partial remission(PR),stable disease(SD),and progressive disease(PD)based on Mac Donald’s criteria.The sensitive group were defined as(CR+PR)and the resistant group were defined as(CD+SD).4.Receiver operating characteristics(ROC)curves were applied to estimate the accuracy of the express intensity of B7-H3 in predicting the early response of concurrent chemoradiotherapy and detect the cut-off values.5.These select patients were divided into high B7-H3 expression group and low B7-H3 expression group according to IHC results.Analysis the effect of B7-H3 expression level combined with MGMT promoter methylation status on prognosis of IDH-wild type GBM.6.Statistical analyses were performed using the SPSS(26.0)software.Chi‐square test or Fisher’s test was applied to evaluate clinical characteristics.Using Kaplan–Meier method to evaluate OS and PFS,and Log‐rank test was employed to compare differences in distributions.p < 0.05 was considered statistical significance.7.Follow up:MRI examinations were performed four weeks after concurrent chemoradiotherapy.Thereafter,patients were followed up every 3 months until December 30,2020.Results1.The MOD value of B7-H3 IHC ranged from 0.0347 to 0.2355(with median MOD of 0.1262).In 65 patients were examined,and 35 of which were B7-H3 low expressed and 30 of which were highly expressed.2.The short-term efficacy was evaluated after one month after chemoradiotherapy,with 19,28,10,8 cases of CR,PR,SD,PD respectively.Values of MOD of resistant group(SD+PD)appeared higher than that of sensitive group(CR+PR)significantly,0.1648±0.0426 vs.0.1164±0.439,(p = 0.001).3.According to the ROC analysis result,a cut-off value of 0.088 for the MOD achieved the highest area under the curve(AUC)at 0.778 and generated the best combination of sensitivity(75.0%)and specificity(77.6%)for distinguishing resistant group and sensitive group.4.Survival analysis showed that the median PFS and median OS in MGMT methylated and unmethylated groups showed no significant difference in the B7-H3 high expression group.(PFS: 7 vs.8 months;OS: 14 vs.15 months,p > 0.05).In contrast,in the B7-H3 low expression group,the median PFS and median OS of MGMT methylated group were significantly higher than that in the non-methylated group(PFS:20vs.10 months;OS: 30 vs.18 months,with both p < 0.05),the difference was statistically significant.ConclusionB7-H3 expression level significant correlates with radiochemosensitivity for patients with IDH-wildtype GBM.Patients with methylated MGMT status and low B7-H3 expression level had good prognosis.B7-H3 could be candidate molecule biomarker for predicating prognosis of GBM.Section 2 Effect of CDKN2 A and MGMT promoter methylation status on prognosis on IDH-wild type GBM Objective1.To detect the status of cyclin-dependent kinase inhibitor 2a(CDKN2A)gene in65 patients with IDH-wild type GBM.2.To investigate the effect of CDKN2 A status combined with MGMT promoter methylation on prognosis of IDH-wild type GBM.Method1.Fluorescence in situ hybridization(FISH)test was used to detect the(CDKN2A)gene status in 65 IDH-wildtype GBM.2.Kaplan-Meier method was adopted to compute the survival time and the logrank test was followed to compare the survival curves.Cox regression model was used to detect prognostic factors.p < 0.05 was considered statistical significance.Result1.CDKN2 A homozygous deletion was noted in 26(40.0%)GBM patients and 39(60.0%)patients were not.2.Survival analysis(1)Patients with CDKN2 A homozygous deletionAmong the 26 patients developed CDKN2 A homozygous deletion,11 cases had MGMT promoter methylated.The median PFS in patients with and without methylated MGMT was 8 and 7 months,respectively,the difference was not significant(p=0.076).The median OS inpatientswith and without methylated MGMT was 13 and10 months,respectively,the difference was not significant(p=0.201).(2)Patients without CDKN2 A homozygous deletionAmong the patients of 39 patients without homozygous deletion of CDKN2 A,22 cases had methylated MGMT status and 17 were not.In patients without CDKN2 A homozygous deletion,there was a significant difference in median PFS between patients with and without methylated MGMT(26 and 12 months,respectively;p=0.006).Accordingly,OS of these patients was significantly different between the two groups.The median OS in patients with and without methylated MGMT was 29 and18 months,respectively,the difference was significant(p=0.0027)ConclusionThe effect of MGMT status on prognosis in patients with IDH-wildtype GBM dependent on CDKN2 A status.更多还原