1、液态活检未来肿瘤领域的液态活检未来肿瘤领域的新星新星内容概括内容概括液态活检的概念液态活检的概念循环肿瘤细胞及其核酸循环肿瘤细胞及其核酸游离肿瘤核酸游离肿瘤核酸液态活检在胶质瘤中的应用液态活检在胶质瘤中的应用循环肿瘤细胞（循环肿瘤细胞（CTCCTC）循环肿瘤细胞循环肿瘤细胞DNADNA（ctDNActDNA）液态活检在胶质瘤中的应用液态活检在胶质瘤中的应用实例：实例：micro-RNAmicro-RNA结果：胶质瘤患者CSF样本中miR-15b、miR-21表达量明显低于对照组。筛选指标：通过检测患者CSF中micro-RNA表达量筛选出胶质瘤特异性指标Micro-RNA实例：实例：micro
2、-RNAmicro-RNA结果：胶质瘤患者CSF中miR-15b、miR-21的相对表达量（Relative expression levels ）均显著低于Control组反向验证：通过检测CSF中miR-15b、miR-21表达量来诊断胶质瘤Micro-RNA实例：实例：micro-RNAmicro-RNA反向验证：针对miR-15b指标ROC分析结果： miR-15b指标曲线下面积AUC=0.96； miR-15b指标REL=0.4作为cut-off值，其敏感性为90%，特异性为94.9% 联合miR-15b、miR-21两指标，诊断准确性更优Micro-RNA实例：实例：micro-R
3、NAmicro-RNA文章NEW FINDINGS： miR-15b富含于胶质瘤患者CSF中，是神经功能紊乱患者组的25倍，PCNSL组的5倍，转移瘤组的17倍，为此，它可作为可靠的胶质瘤诊断指标；miR-15b通过调节细胞周期，参与胶质瘤的发生过程。它可能是一个抑癌基因，在细胞周期G1期作用于Cyclin E1(CCNE1)；miR-21同样高表达于胶质瘤患者CSF中，它是一个肿瘤基因，可以抑制肿瘤细胞凋亡；它可以靶向作用于一些抑癌基因，如PTEN、PDCD4等；它可以直接下调TIMP3，增加肿瘤细胞侵袭性；Micro-RNA实例：实例：ProteinsProteins 方法：从PubMed
4、 MEDLINE上文献检索相关研究蛋白； 标本：收集29例胶质瘤患者23份CSF样本，3份肿瘤囊液； Proteins检测方法：ELISA；Proteins实例：实例：ProteinsProteins 14.7% were signaling molecule, 8.8% were cell adhesion molecule, and 8.8% were lyase)；Proteins实例：实例：ProteinsProteins 40.7% binding, 25.9% catalytic activity, and 14.8% structural molecule activity),
5、and 7 molecular function groups；Proteins实例：实例：ProteinsProteins 16.7 % cellular process, 14.8 % developmental process, and 14.8 % metabolic process， 12 biological process groups； Proteins实例：实例：ProteinsProteins It is not surprising to find that most proteins were located at the extracellular space (52
6、 %) ；Proteins实例：实例：ProteinsProteins GBM囊液中蛋白含量显著高于其他胶质瘤CSF中蛋白含量； LGG中CSF蛋白含量低于对照组、AA/AO组、GBM组；Proteins实例：实例：ProteinsProteins GBM囊液中GAL蛋白含量显著低于对照组、LGG组、AA/AO组、GBM组的CSF中的含量，但LGG组、AA/AO组、GBM组间无显著差异；实例：实例：ProteinsProteins GBM囊液中WNT4蛋白含量显著低于对照组、LGG组、AA/AO组、GBM组的CSF中的含量，但LGG组、AA/AO组、GBM组间无显著差异；Proteins实例：
7、实例：ProteinsProteins GBM囊液中HSPA5蛋白含量显著低于对照组、LGG组、AA/AO组、GBM组的CSF中的含量，但LGG组、AA/AO组、GBM组间无显著差异；Proteins实例：实例：ProteinsProteins GBM囊液中IL-6蛋白含量显著高于于对照组、LGG组、AA/AO组、GBM组的CSF中的含量； 在CSF中IL-6含量随胶质瘤级别增高，含量增加；Proteins实例：实例：ProteinsProteins文章NEW FINDINGS： IL-6可能是GBM患者CSF中特异性分子标记物；IL-6参与免疫应答、急性炎症反应、造血等过程，它可能在机体防御
8、过程中起重要作用；有证据表明IL-6是致瘤性活动中重要的炎症介质；IL-6通过炎症信号通路，如JAK2-STAT3 and Jagged-Notch等，可增加GBM肿瘤细胞侵袭、增殖能力，并减少GBM肿瘤细胞的凋亡；为此，IL-6或许可以成为潜在的药物设计靶点；ProteinsMMP2 and MMP9对原发或转移脑肿瘤（其中包括29名胶质瘤患者）实验组和对照组总共66名患者的脑脊液样本进行电泳，检测MMP2和MMP9的前体形式（pMMP2/9)和活化形式（MMP2/9),统计阳性百分比MMP2 and MMP9健康人细胞学检查阴性的多形性胶质瘤患者细胞学检查阳性的多形性胶质瘤患者多形性成胶质
9、细胞瘤软脑膜肿瘤患者MMP9的衍生物MMP9MMP9和TIMP复合物MMP2脑脊液电泳结果文章发现l 电泳结果显示,所有66名被检测者MMP2前体均阳性，而MMP9前体只在脑肿瘤（无论原发还是转移）患者脑脊液中发现，而MMP9衍生物（250kDa gelatinase）仅在胶质瘤和部分脑转移瘤患者中被发现；l 高级别胶质瘤患者脑脊液中MMP2和MMP9水平明显增加；l MMPs可用作中枢神经系统肿瘤诊断和监测进展的标志物。VEGF and FGF-生存期高级别和低级别胶质瘤以及脑积水病人脑脊液中VEGF 和FGF-的水平以及生存曲线文章发现l 高级别胶质瘤患者脑脊液中VEGF and FGF-
10、水平明显高于低级别胶质瘤或脑水肿的病人；l 胶质瘤患者脑脊液中VEGF 和FGF-水平与肿瘤血管化有关，并与患者的生存期成负相关；l 胶质瘤患者脑脊液中VEGF 和FGF-水平可以用做判断肿瘤血管化的辅助标志物，从而帮助预测患者的生存期；实例：实例：ProteinsProteinsCTCs实例：实例：ProteinsProteinsCTCs The dogma that GBM spread is restricted to the brain was challenged by reports on extracranial metastases after organ transplant
11、ation from GBM donors. We identified circulating tumor cells (CTCs) in peripheral blood (PB) from 29 of 141 (20.6%) . GBM patients by immunostaining of enriched mononuclear cells with antibodies directed against glial fibrillary acidic protein (GFAP). Tumor cell spread was not significantly enhanced
12、 by surgical intervention. The tumor nature of GFAP-positive cells was supported by the absence of those cells in healthy volunteers and the presence of tumorspecific aberrations such as EGFR gene amplification and gains and losses in genomic regions of chromosomes 7 and 10. Release of CTCs was asso
13、ciated with EGFR gene amplification, suggesting a growth potential of these cells. We demonstrate that hematogenous GBM spread is an intrinsic feature of GBM biology.实例：实例：ProteinsProteinsCTCs实例：实例：ProteinsProteinsCTCs 用来鉴别假性进展与肿瘤复发； 评价放射治疗效果；实例：实例：ProteinsProteinsCTCs实例：实例：ProteinsProteinsCTCsGBMs
14、are locally invasive within the brain but rarely metastasize to distant organs, exemplifying the debate over “seed” versus “soil.” We demonstrate that GBMs shed CTCs with invasive mesenchymal characteristics into the circulation. Rare metastatic GBM lesions are primarily mesenchymal and show additional mutations absent in the primary tumor.
