鼻咽癌患者外周血循环肿瘤细胞的检测及其与预后的相关性

目的:利用抗体标记和流式细胞仪联合方法检测鼻咽癌外周血中的循环肿瘤细胞(circulating tumor cells,CTCs)数量,分析评价CTCs与患者临床特征和预后之间的相关性.方法:针对福建省肿瘤医院2012年7月至2014年5月收治的67例鼻咽癌首诊患者以及10例健康志愿者,抽取外周血10ml,提取其中的单个核细胞.依次应用抗Epcam、抗CD45以及抗CK单克隆抗体对上述提取的单个核细胞进行抗体标记,将Epcam阳性、CD45阴性以及CK阳性的细胞作为鼻咽癌患者的CTCs,应用流式细胞仪检测其在外周血中的存在情况,分析其与患者临床特征及预后之间的关系.结果:鼻咽癌组外周血中的CTCs明显高于健康志愿者组,差异具有统计学意义(P<0.05).单因素分析结果表明,鼻咽癌患者外周血CTCs水平与进展情况(P=0.04)、N分期(P=0.001)以及临床分期(P=0.035)均显著相关.Logistic多因素回归分析结果则显示,仅患者的N分期与CTCs水平具有相关性(P=0.003),CTCs阳性患者的N分期更晚.Kaplan-Meier生存分析显示,CTCs阳性患者的无进展生存期(progression free survival,PFS)要显著短于CTCs阴性患者(P=0.029).结论:鼻咽癌患者外周血中CTCs阳性与患者N分期相关,且提示预后不良,可能成为鼻咽癌患者预后评估的一个辅助指标.     

晚期卵巢癌患者外周血循环肿瘤细胞的检测及其临床意义

目的:探讨晚期卵巢癌患者外周血中循环肿瘤细胞(CTC)的检测及其临床意义.方法:选择2014年6月至2016年6月永州市中心医院收治的60例晚期卵巢癌患者作为观察组,收集同期本院的30例健康志愿者作为对照组.分别抽取外周血7.5 ml,采用流式细胞仪检测卵巢癌患者一线方案化疗2周期前后及对照组外周血中EpCAM、CK19同时(+)且CD45(-)的CTC的变化情况,同时分析其与临床特征及化学疗效的关系以及比较化疗前后CTC数目评价与RECIST疗效评价标准之间的关系,并随访患者的无进展生存期(PFS),分析患者治疗过程中CTC数目的变化与患者PFS的关系.结果:观察组中42例检测到CTC,阳性率70%,对照组未检测到CTC,两组比较,差异具有统计学意义(P<0.05).CTC阳性表达情况与卵巢癌患者的分化程度及CA125表达相关(P<0.05),与患者的年龄、肿瘤直径、腹水情况均无相关性(P>0.05).观察组中化疗前CTC阳性表达42例,阳性率为70%,化疗2周期后卵巢癌患者CTC阳性表达18例,阳性率为30%,两组差异具有统计学意义(P<0.05).化疗2周期后,CTC数目评价与RECIST疗效评价标准之间差异无统计学意义(P>0.05),且两种方法的吻合度较高(k=0.479).运用相关分析显示,CTC表达与化学疗效呈负相关(r=-0.223,P=0.009).化疗2周期后外周血中循环肿瘤细胞数目<5,其无进展生存期为13.2(11.0~15.1)个月,外周血中循环肿瘤细胞数目≥5,其无进展生存期为6.4(3.8~9.2)个月,两组差异具有统计学意义(P<0.05).结论:CTC阳性表达与卵巢癌患者的分化程度及CA125表达相关,与化疗疗效呈负相关,CTC的表达情况可以预测卵巢癌患者的化疗疗效及预后.     

Detection of breast cancer cells in the peripheral blood is positively correlated with estrogen-receptor status and predicts for poor prognosis

We investigated whether detection of cytokeratin-positive (CK+) cells in the peripheral blood (PB) of breast cancer patients before chemotherapy could be a prognostic factor. Blood from a total of 92 breast cancer patients was evaluated for the presence of CK+ cells. Blood samples were collected before chemotherapy. Patients entered in the study included: neoadjuvant (n = 25), adjuvant (n = 42) and metastatic (n = 25). Blood samples (10 ml) were centrifuged using a double density-gradient to recovering the mononuclear cell (MNC) and granulocyte cell (GC) fractions. Subsequently, positive immunomagnetic cell separation was carried out to isolating CK+ cells. The enriched cell fraction was cytocentrifuged and then immunocytochemically labeled using an anti-cytokeratin antibody. Our results indicated that breast tumor cells sediment with both MNC and GC fractions. We therefore recommend examination of both fractions in all enrichment protocols. CK+ cells in PB were identified in 57 of 92 (62%) patients when MNC and GC fractions were assessed (range = 1-61 cells, median = 8). No CK+ cells were detected in blood samples of 16 healthy donors. There were significant differences in the presence of CK+ cells according to estrogen receptor expression (p = 0.049), and lymph node status (p = 0.033), but not to the age, menopausal status, type of patient (neoadjuvant, adjuvant or metastatic), TNM stage, histological type, progesterone receptor expression, c-erbB2 expression, p53 expression or Ki67 expression. Regarding the relationship between tumor size (T) and the presence of CK+ cells, a borderline significant trend was observed (p = 0.07). The median follow-up of the patients was 21 months and statistical analysis (Kaplan-Meier analysis) showed that using the method we present, the detection of CK+ cells in PB before starting the chemotherapy in breast cancer patients was significantly correlated with both progression-free survival (p = 0.058) and overall survival (p = 0.003). In conclusion, the present study suggests that detection of CK+ cells in PB before chemotherapy might identify breast cancer patients with poor prognosis.     

肺癌患者外周血循环内皮细胞和VEGF检测临床意义的探讨

目的:探讨肺癌患者外周血循环内皮细胞(CECs)包括活性循环内皮细胞(aCECs)与VEGF之间的关系及其在肺癌中的临床意义。方法:用流式细胞仪对70例肺癌患者(NSCLC64例)化疗前后及30例健康人外周血CECs及aCECs进行检测,同时采用ELISA方法检测肺癌患者化疗前后VEGF水平。结果:NSCLC患者外周血CECs及aCECs的数量明显高于健康人群(P=0.000)。不同病理类型肺癌之间CECs及aCECs的数量差异无统计学意义(NSCLC与SCLC,P=0.878、0.936;鳞癌与腺癌,P=0.786、0.658)。CECs及aCECs的数量与VEGF之间无明显相关性(P=0.697、0.631)。治疗后不同疗效间肺癌患者VEGF水平比较,PD均高于CR+PR及SD(P=0.009、0.031)。NSCLC及SCLC化疗后aCECs的数量高于化疗前(P=0.031、0.029);SD及PD患者CECs、aCECs的数量高于化疗前(P=0.024、0.014;P=0.036、0.028);对治疗后不同疗效肺癌患者CECs及aCECs水平比较,aCECs数量PD、SD均高于CR+PR(P=0.022、0.009)。结论:VEGF联合CECs及aCECs检测可以作为很好的预测性指标评估肺癌的血管生成状况及判断预后。     

High‐resolution imaging for the detection and characterisation of circulating tumour cells from patients with oesophageal,hepatocellular, thyroid and ovarian cancers

Interest has increased in the potential role of circulating tumour cells in cancer management. Most cell‐based studies have been designed to determine the number of circulating tumour cells in a given volume of blood. Ability to understand the biology of the cancer cells would increase the clinical potential. The purpose of this study was to develop and validate a novel, widely applicable method for detection and characterisation of circulating tumour cells. Cells were imaged with an ImageStream^X imaging flow cytometer which allows detection of expression of multiple biomarkers on each cell and produces high‐resolution images. Depletion of haematopoietic cells was by red cell lysis, leukocyte common antigen CD45 depletion and differential centrifugation. Expression of epithelial cell adhesion molecule, cytokeratins, tumour‐type‐specific biomarkers and CD45 was detected by immunofluorescence. Nuclei were identified with DAPI or DRAQ5 and brightfield images of cells were collected. The method is notable for the dearth of cell damage, recoveries greater than 50%, speed and absence of reliance on the expression of a single biomarker by the tumour cells. The high‐quality images obtained ensure confidence in the specificity of the method. Validation of the methodology on samples from patients with oesophageal, hepatocellular, thyroid and ovarian cancers confirms its utility and specificity. Importantly, this adaptable method is applicable to all tumour types including those of nonepithelial origin. The ability to measure simultaneously the expression of multiple biomarkers will facilitate analysis of the cancer cell biology of individual circulating tumour cells.     

乳腺癌患者外周血循环肿瘤细胞 MiR -21检测的临床意义

目的：研究乳腺癌患者血浆中的 MiR -21表达水平，初步探讨其在乳腺癌中作为循环肿瘤细胞（cir-culating tumor cells，CTCs）的临床意义。方法：采用基于 SYBR Green I 染料荧光分析的茎环实时定量逆转录聚合酶链式反应（qRT - PCR）分别检测40例乳腺癌患者及15例正常对照者外周血中 MiR -21的表达水平，了解 MiR -21表达与临床病理指标的关系。结果：乳腺癌患者血浆中的 MiR -21表达水平明显高于正常对照者（P ﹤0.01）。MiR -21表达与患者月经状况、肿瘤大小、雌激素受体（ER）、孕激素受体（PR）和人类表皮生长因子受体2（HER -2）表达均无明显关系（P ﹥0.05），与 TNM 分期、淋巴结转移相关（P ﹤0.05）。结论：茎环 qRT - PCR 是一种快速、准确、定量检测 MiR -21的方法，MiR -21是一个能有效辅助乳腺癌诊断和监测 CTCs 的潜在标志物。     

分化型甲状腺癌NIS表达与外周血CT C的相关性研究

目的：探讨分化型甲状腺癌钠／碘同向转运体（sodium／iodide symporter,NIS）与外周血循环肿瘤细胞（circulating tumor cell,CTC）的相关性。方法收集分化型甲状腺癌172例。通过免疫组织化学SP法及流式细胞术,对甲状腺癌组织NIS表达及外周血CTC 阳性率进行检测分析。结果分化型甲状腺癌组织NIS 表达76例（44．2％）,外周血CTC阳性63例（36．6％）。淋巴结 N0组 NIS 阳性表达较 N1组多,差异有统计学意义（χ2＝6.015,P＝0.014）,N0组CTC阳性率低于N1组,差异亦有统计学差异（χ2＝14.035,P＝0.001）。在N0组及N1组中NIS表达与CTC阳性率均呈负相关（r＝－0.383,r＝－0.610,均P＜0.01）。各病理亚型之间NIS表达高分化型多于中间分化型,差异有统计学意义（χ2＝7.897,P＝0.005）,CTC阳性率高分化型较中间分化型低,差异无统计学意义（χ2＝1.455,P＝0.228）。高分化型及中间分化型中 NIS 表达与 CTC 阳性率均呈负相关（r＝－0.591,r＝－0.443,均P＜0.01）。结论分化型甲状腺癌组织NIS表达与外周血CTC阳性率呈负相关。     

膀胱癌患者外周血中循环肿瘤细胞的监测及临床意义研究

目的研究膀胱癌患者外周血中循环肿瘤细胞(CTC)数目,探讨其对膀胱癌复发和转移的早期监测、临床疗效评价及预后判断的意义。方法选取87例膀胱癌患者及40例同期膀胱良性病变患者作为研究对象,根据肿瘤浸润深度将87例膀胱癌患者进一步分为非肌层浸润组(55例)和肌层浸润组(32例),膀胱良性病变患者作为对照组(40例)。采用上皮细胞黏附分子(EpCAM)芯片分选法检测外周血中CTC,比较各组治疗前及治疗过程中的CTC数量,比较不同CTC数量患者治疗后1年复发率。结果治疗前,对照组患者外周血中未检出CTC,非肌层浸润组患者CTC检出数为(9.38±11.35)/10 ml,明显少于肌层浸润组患者的(23.28±11.74)/10 ml,差异有统计学意义(P﹤0.01)。治疗过程中两组膀胱癌患者CTC检出数均逐渐减少,差异均有统计学意义(P﹤0.01)。未检出CTC的非肌层浸润组患者1年复发率低于检出CTC的非肌层浸润组患者,差异有统计学意义(P﹤0.05);CTC检出数为(0～20)/10 ml及≥21/10 ml的肌层浸润组患者的1年复发率比较,差异无统计学意义(P﹥0.05)。结论外周血中的CTC是膀胱癌预后不良的重要标志,对膀胱癌临床疗效评价、预后判断有重要价值。     

外周血循环肿瘤细胞检测的研究进展

Metastasis usually presents in the different courses of tumor diseases and associates with poor prognosis. Circulating tumor cells (CTC) are considered to be essential for conforming metastasis and can be detected in tumor patients’ peripheral blood. CTC are not easily detected by conventional cytology methods because of their low frequency in peripheral blood. Our article reviews the recent research results on this subject, and also discusses the problems and prospects in this area.     

Clinical implications of microRNAs in liver cancer stem cells

The prognosis of patients diagnosed with hepatocellular carcinoma (HCC) is often dismal, mainly due to late presentation, high recurrence rate, and frequent resistance to chemotherapy and radiotherapy. Accumulating evidence on the differential microRNA (miRNA) expression patterns between non-tumor and HCC tissues or between liver cancer stem cells (CSCs) and non-CSC subsets and the significant clinical implications of these differences suggest that miRNAs are a promising, non-invasive marker for the prognosis and diagnosis of the disease. This perspective article summarizes the current knowledge of miRNAs in liver CSCs and highlights the need for further investigations of the role of miRNAs in regulating liver CSC subsets for possible future clinical applications.     

左右侧肺癌患者外周血循环肿瘤DNA突变特征分析

目的:探究左右侧肺癌患者外周血循环肿瘤DNA(ctDNA)突变特征及导致治疗响应与预后差异产生的潜在分子机制.方法:本研究对14例肺癌患者的外周血标本ctDNA行高通量测序,并利用生物信息学方法分析左右侧肺癌组间ctDNA突变特征、蛋白质结构与功能及信号调控网络等的变化.结果:发现左侧肺癌组中检出突变种类及多重突变数量...     

分化型甲状腺癌碘131治疗后的外周血细胞改变

甲状腺癌是近五年来发病率增长最为快速的肿瘤之一,中国每年新发甲状腺癌患者将近50 000名,并呈上升趋势。核医学作为非手术治疗方式,在普外科应用广泛,且绝大部分病人治疗效果反应好,不良反应发生率低。有研究显示,长期大剂量的碘辐射会诱发骨髓抑制,导致全血细胞下降,甚至诱发白血病。本文旨在总结近年来分化型甲状腺癌术后患者在接受放射性碘131治疗后,红细胞、白细胞、淋巴细胞及血小板等外周血细胞以及细胞因子数量的变化,以探究其是否具有危害性。结果表明甲状腺癌切除术后,应用放射性碘131治疗,是一种安全有效的治疗方式。     

乳腺癌患者甲状腺功能的相关研究

目的：探讨乳腺癌患者的甲状腺功能情况及与发病间的关系,乳腺癌患者自身免疫与非自身免疫甲状腺疾病的发病情况。方法：收集2008年1月至2013年1月我院诊治的乳腺癌病例100例、乳腺良性疾病患者100例,正常对照病例100例。以放射免疫检测法测血清 FT3、FT4、TSH、TG －Ab、TPO －Ab 水平;通过临床检查、B 型超声、甲状腺素及抗体水平检查、细针穿刺活组织检查等对比甲状腺疾病的发病情况,并分析与乳腺癌的关系。结果：乳腺癌组患者血清 FT4水平（16．56±0．45）pmol／L 和 TPO －Ab 水平（51．44±22．59） U ／ml 虽均在正常范围,但其 FT4水平显著低于乳腺良性疾病组和正常对照组（P ＝0．023,P ＝0．017）,其 TPO－Ab 水平高于乳腺良性疾病组和正常对照组（P ＝0．013,P ＝0．002）。乳腺癌组非自身免疫与自身免疫甲状腺疾病的发病率高于对照组[36％（36／100）vs 12％（12／100）,P ＝0．004;22％（22／100）vs 14％（14／100）, P ＝0．001],乳腺良性疾病组与对照组相近。乳腺癌 FT4降低与乳腺癌的肿块大小、肿瘤分期、淋巴结转移有关联（P ＜0．05）。与年龄、病理类型、ER、PR 表达无关联（P ＞0．05）;乳腺癌 TPO －Ab 高表达与乳腺癌的肿块大小、肿瘤分期、淋巴结转移、ER、PR 表达有关联（P ＜0．05）,与年龄、病理类型无关联（P ＞0．05）。追踪条件匹配的40例乳腺癌放化疗及内分泌治疗,发现与放射肿瘤及内分泌治疗有关联（P ＜0．05）。结论：甲状腺功能变化和乳腺癌的发生发展有一定联系。甲状腺激素和自身抗体水平有助于临床监测和预后判断,乳腺癌患者的非自身免疫与自身免疫甲状腺疾病的发病率高于非乳腺癌者。乳腺癌患者 FT4水平降低及 TPO －Ab 高表达可反映乳腺癌的严重程度。     

肺癌患者外周血循环中癌细胞的流式细胞仪分析

目的：用流式细仪分析肺癌患者外周血循环中的癌细胞,方法：外周血经Ficoll梯度离心分离单核细胞组分,后者用CD45、细胞角蛋白（CK）及肺癌特异性抗体（2F／S5A）染色后,应用流式细胞仪检测CD45^-CK^ /S5A^ 细胞。结果检测了165例肺癌患者外周血,发现50例（30．3％）患者外周血癌细胞呈阳性。其中非小细胞肺癌阳性率为30．67％（45／150）,小细胞肺癌为33．33％,阳性率为与患者的病理分期有显著相关性（P＜0．05）。结论：利用流式细胞仪分析检测癌患者外周血循环中的癌细胞不驻有助于临床病理分期,而且对预测肺癌转移潜能具有应用价值。