循环肿瘤DNA监测早期肺癌术后微小残留病灶的研究进展

循环肿瘤DNA(circulating tumor DNA,ctDNA)已经应用于监测转移性非小细胞肺癌（non-small cell lung cancer,NSCLC）的治疗。近来研究者开始关注ctDNA预测早期NSCLC患者根治术后微小残留病灶(minimal residual disease,MRD)的价值。全文总结目前常用的ctDNA检测方法,并介绍了其在早期肺癌术后评估MRD和复发预警方面的应用进展。     

基于循环肿瘤DNA的微小残留病灶检测在结直肠癌预后中的应用进展

辅助化疗已经被广泛用于治疗Ⅱ/Ⅲ期结直肠癌患者,然而是否对某些特定患者进行辅助化疗仍然存在争议。基于TNM分期系统评估患者个体复发风险的准确性有待提高。研究显示患者术后体内的循环肿瘤DNA(ctDNA)状态是一个非常有效的预后标志物。术后ctDNA反映了微小残留病灶的存在,并将从根本上改变了复发风险和辅助治疗疗效的评估方式。本文主要介绍基于ctDNA的微小残留病灶检测在结直肠癌预后中的临床应用和未来发展方向。     

转移性乳腺癌循环肿瘤细胞的上皮间质转化水平对化疗疗效的预测意义

目的 检测转移性乳腺癌患者外周血循环肿瘤细胞（CTCs）不同亚型相关基因的表达,探讨其对化疗疗效的预测意义。方法 应用免疫磁性分选（MACS）技术联合逆转录 聚合酶链反应（RT-PCR）法检测58例转移性乳腺癌患者和10例健康人外周血中CTCs上皮型标志物上皮角蛋白（CK18、CK19）和间质型标志物（波形蛋白、纤连蛋白）mRNA的表达。分析上皮型标志物及间质型标志物表达与不同亚型乳腺癌之间的关系,并分别评估具有不同表型CTCs患者之间疗效的差异。结果 在10例健康志愿者的血液样本中,未检测出CK18、CK19、波形蛋白和纤连蛋白mRNA的表达。在58例转移性乳腺癌患者的血液样本中,检测出36例（62.1％）上皮角蛋白表达,19例（32.8％）间质型标志物表达。Luminal A组和HER-2阳性组的上皮型标志物阳性表达率高于三阴性乳腺癌组（P=0.008）,而间质型标志物阳性表达率则低于三阴性乳腺癌组（P＜0.001）。根据不同标志物的表达情况,将患者分为CKs+／EMT-组、CKs-／EMT-组、CKs+／EMT+组和CKs-／EMT+组,4组有效率依次为76.7％、55.6％、33.3％和15.4％,差异有统计学意义（P=0.002）。间质型标志物阴性者的化疗有效率高于间质型标志物阳性者（71.8% vs. 15.8%,P=0.000）。结论 转移性乳腺癌患者中部分CTCs将发生上皮间质转化而丢失上皮型细胞的表型,获得间质型细胞表型。间质型CTCs因具有更强的抵抗化疗药物的能力而存活,这可能是三阴性乳腺癌疗效不佳的原因之一。     

循环肿瘤细胞的异质性在转移性乳腺癌中的作用研究进展

循环肿瘤细胞（CTC）的计数可以预测转移性乳腺癌的预后,但其改善患者预后的能力在临床试验中尚未得到证实。目前研究专注于CTC的分子表征,作为肿瘤组织的“替代物”以非侵入性地方式评估癌症基因组表达及其在治疗过程中的演变。CTC中存在上皮-间质转化过程（EMT）,其特点为上皮标志物的缺失。EMT过程可以存在于侵袭性及耐药性较强的细胞,其计数和表征,能够引起肿瘤的复发和进展,具有较高的临床价值。本文深入探讨循环肿瘤细胞的异质性及在转移性乳腺癌上皮-间质转化过程中的作用。使其成为乳腺癌患者监测转移和预后的常规的检测指标,并有助于明确转移的机制,更有望发现乳腺癌转移治疗的新靶点。     

PCR检测降钙素基因高甲基化在白血病微小残留病灶诊断中的价值

为探讨甲基化分析在白血病微小残留病灶（ＭＲＤ）中的价值，用酶切加ＰＣＲ技术研究３１例初诊期急性髓细胞白血病（ＡＭＬ），１４例对照者以及在１８个月中动态观察５例ＡＭＬ治疗前后降钙素（ＣＴ）基因高甲基化的变化。３１例初诊期ＡＭＬ中２５例，１４例对照者中的０例呈现ＣＴ基因高甲基化（Ｐ〈０．０１）。动态观察的５例ＡＭＬ，全部初诊期及完全缓解时的４例呈现ＣＴ基因高甲基化，其中３例分别于完全缓解后３个月、５个     

乳腺癌循环肿瘤细胞研究及其检测进展

乳腺癌是女性发生率最高的恶性肿瘤之一,其引起死亡的主要原因是乳腺癌的复发转移.转移灶的形成是一个连续复杂的过程,从原发灶生长的早期即已开始.肿瘤细胞进入血液循环是乳腺癌发生转移的基础.经过多种复杂机制最终形成转移灶.因此,检测循环肿瘤细胞(CTC)对预测转移有重要作用.由于外周血中CTC数量稀少,增加了其检测的难度.目前对CTC的检测主要分为基于抗体的技术、核酸技术以及联合检测、系统检测等方式,随着试验方法的改进和新技术的不断出现,CTC检测的敏感度和特异性有较大提高.然而缺乏高特异性的标志物仍是CTC检测面临的最大问题.乳腺癌患者CTC水平与肿瘤的分期相关,其水平的升高提示有更高转移的可能.预示患者的预后较差.与传统的组织学及影像学检查相比,CTC检测有更高的可重复性和敏感度,并能更早地提供预后信息.与骨髓微转移肿瘤细胞检测相比,CTC检测具有创伤小、可连续多次检测等特点.CTC检测更重要的意义是能够帮助指导临床个体化治疗方案的制定.通过对CTC连续的检测可以及早地评估治疗效果,据此及时转变为更有效的治疗方案,提高患者的生存率.本文对乳腺癌循环肿瘤细胞的研究、检测技术和临床应用的新进展进行综述.     

乳腺癌患者循环肿瘤细胞检测研究进展

0引言肿瘤转移是乳腺癌治疗失败的首要原因。乳腺癌患者循环血中的微转移经常规病理检查和影像检查如X线、CT等均难以发现。早在1999年,Ghos-     

局部晚期非小细胞肺癌微小残留病灶的研究进展北大核心CSCD

局部晚期不可手术非小细胞肺癌(NSCLC)的标准治疗为同步放化疗(CCRT)后巩固免疫治疗,但后续可能复发。影像学检查只能被动识别复发、无法提前预测风险,基于循环肿瘤DNA(ctDNA)检测的微小残留病灶(MRD)则成为新兴的肿瘤标志物。研究表明,ctDNA在NSCLC放(化)疗后先升后降,不仅可以预测复发,还可能指导巩固免疫治疗、评估疗效和判断预后,甚至有助于探索其他有效的治疗方案。目前将ctDNA-MRD用于指导巩固治疗相关的临床研究较少、样本量较小、证据等级较低,其临床价值仍需前瞻性随机研究进一步验证。     

Minimal residual disease and circulating tumor cells in breast cancer

Tumor cell dissemination in bone marrow or other organs is thought to represent an important step in the metastatic process. The detection of bone marrow disseminated tumor cells is associated with worse outcome in early breast cancer. Moreover, the detection of peripheral blood circulating tumor cells is an adverse prognostic factor in metastatic breast cancer, and emerging data suggest that this is also true for early disease. Beyond enumeration, the characterization of these cells has the potential to improve risk assessment, treatment selection and monitoring, and the development of novel therapeutic agents, and to advance our understanding of the biology of metastasis.     

The minimal residual disease (MRD) in hematological malignancies]

Molecular genetic and cytoimmunological markers have been applied for the detection of minimal residual disease (MRD) in hematological malignancies. These markers include surface markers or rearranged T-cell receptor and immunoglobulin genes in the lymphoid malignancies and fused genes associated with chromosomal translocations such as BCR-ABL in t(9;22) or PML-RAR alpha in t(15;17) in myeloid malignancies. The expression of the WT1 gene is recognized as the universal tumor marker for a wide variety of hematological malignancies. Using these sensitive markers, a tumor cell in 10,000 to 1,000,000 normal cells can be detected. By examining a large number of patients, it has been shown that the MRD in the early phase of chemotherapy has a correlation with the prognosis of childhood ALL. Based on these observations, a new strategy of chemotherapy in which the post-remission therapy is modified based on the MRD results has begun. The amount of tumor cells contaminated in the autologous stem cell grafts in ALL patients might be related to the prognosis. The diagnosis of MRD will be used as an important routine examination in chemotherapy for leukemia/lymphoma patients.     

乳腺癌循环肿瘤细胞检测及其研究进展

循环肿瘤细胞(CTC)为肿瘤发生转移的重要原因.检测CTC能早期发现肿瘤的转移及复发.在乳腺癌中,CTC检测与其他检测方式相比有较高的可重复性和敏感性,并能更早地提供预后信息.     

Detection of minimal residual disease (MRD) after bone marrow transplantation (BMT) by multi-parameter flow cytometry (MPFC)

Multi-parameter flow cytometry (MPFC) was used to detect minimal residual disease (MRD) following bone marrow transplantation (BMT) in 21 patients. Bone marrow (BM)was analyzed pre-transplant and 3-4 months post-BMT while the patients were in clinical and morphological remission. MRD was detected by identifying cells with aberrant antigen expression and/or leukemia-associated phenotype (LAP) using MPFC. Prior to BMT, 8 out of 21 patients exhibited normal antigen expression based on normal BM samples while 13 BM aspirates had abnormal MPFC. Pre-BMT MPFC was abnormal in all 10 patients who were not in complete remission (CR) (>5% blasts in BM) as well as 3 patients acute lymphoblastic leukemia (ALL) who were in CR. In BM from ALL patients, an abnormal uniform B cell population was observed however antigen expression patterns varied greatly between patients. BM from acute myeloblastic leukemia (AML) patients showed an abnormal distribution of CD34+ cells. In addition, a correlation was observed between pre-BMT cytogenetics and MPFC. Only 2 out of 8 (25%) patients with normal MPFC pre-autologous bone marrow transplantation (ABMT) relapsed (AML), while 6 out of 13 (46%) patients with abnormal pre-BMT MPFC relapsed including 2 out of 3 patients who were transplanted in clinical CR. Pre-BMT MPFC may thus be an effective tool for detection of MRD by detection of a pre-transplant MPFC abnormality.     

循环肿瘤细胞及循环肿瘤DNA检测在乳腺癌中的研究进展

乳腺癌是中国女性最常见的癌症,其早期检测、个体化治疗及实时疾病监测一直是科研及临床工作者关注的重点。癌症患者血液中循环肿瘤细胞(CTC)和循环肿瘤DNA(ctDNA)的检测微创、简便,易实时获取信息。目前,有关CTC富集鉴定方法多种多样,CTC及ctDNA检测的敏感度和特异度较前有所提高,未来研究面临的最大挑战为肿瘤细胞特异性标志物的开发以及ctDNA临床意义的研究。CTC及ctDNA在基础研究、检测方法、CTC培养、早期肿瘤检测、个体化治疗及临床预后等方面的研究均有开展。笔者对乳腺癌CTC及ctDNA的研究进展作一总结。     

CD49f-based selection of circulating tumor cells (CTCs) improves detection across breast cancer subtypes

Circulating tumor cells (CTCs) can be enumerated using CellSearch, but not all breast cancer subtypes, specifically those with epithelial-mesenchymal transition (EMT) characteristics, sufficiently express the enrichment (EpCAM) and selection (CK8/18/19) markers used in this method. While CD146 can detect EpCAM-negative CTCs, we here evaluated the value of various cytokeratins and CD49f to detect CK8/18/19-negative CTCs. The tested cytokeratins provided no substantial benefit, but adding CD49f to CK8/18/19 as a selection marker resulted in improved recovery of normal-like cell lines.     

CD49f-based selection of circulating tumor cells (CTCs) improves detection across breast cancer subtypes

Dibenzo[def,p]chrysene (DBC) is a transplacental carcinogen in mice (15 mg/kg; gestation day (GD) 17). To mimic residual exposure throughout pregnancy, dams received four smaller doses of DBC (3.75 mg/kg) on GD 5, 9, 13 and 17. This regimen alleviated the previously established carcinogenic responses in the thymus, lung, and liver. However, there was a marked increase in ovarian tumors (females) and hyperplastic testes (males). [¹⁴C]-DBC (GD 17) dosing revealed transplacental distribution to fetal tissues at 10-fold lower concentrations than in paired maternal tissue and residual [¹⁴C] 3 weeks post-dose. This study highlights the importance of developmental stage in susceptibility to environmental carcinogens.