CTCs在膀胱癌转移及预后评估中应用的研究

目的 检测膀胱癌患者外周血循环肿瘤细胞(circulating tumor cells, CTCs)的计数及分型,研究其在膀胱癌的转移及预后评估中的临床应用价值. 方法 采用CanPatrolTM二代CTCs检测技术检测外周血CTCs计数及分型.41例经病理确诊的膀胱癌患者纳入研究,患者年龄33~83岁,男37例,女4例.肿瘤TNM分期:T1期15例、T2期12例、T3期9例、T4期5例、N1期6例、M1期3例.肿瘤组织病理分级:高级别膀胱癌30例、低级别膀胱癌11例. 结果 41例膀胱癌患者总CTCs阳性率为68.29%(28/41),6例淋巴结转移患者中检出CTCs阳性2例,3例远处转移患者中检出CTCs阳性1例.本研究共检出CTCs 136个,其中上皮型CTCs 36个(26.5%)、混合型CTCs 70个(51.5%)、间质型CTCs 30个(22.0%);间质化CTCs 100个,占73.5%.30例高级别膀胱癌中CTCs检出20例,占66.7%,11例低级别膀胱癌中CTCs检出7例,占63.6%.13例CTCs阴性患者中,无淋巴结转移10例(69.23%),淋巴结转移3例(30.76%).T1G3患者间质型CTCs阳性率为60.0%,CTCs总数≥3的患者比例为60.0%;T1G1~2患者间质型CTCs阳性率为20.0%,CTCs总数≥3的患者比例为20.0%.41例中29例患者术后获得随访,随访12~41个月,平均(13±2)个月,死亡8例.术后总生存率为72.41%,其中CTCs阳性者生存率为77.77%(14/18),CTCs阴性者生存率为63.60%(7/11). 结论 T1G3患者的间质型CTCs阳性率及CTCs总数≥3的患者比例明显高于T1G1~2患者.CTCs的总数及间质型CTCs的个数能够在一定程度上预测膀胱癌的恶性程度,并指导非肌层浸润性膀胱癌患者的早期治疗.CTCs的阳性检出率与膀胱癌的TNM分期、淋巴结转移、近期生存率之间未发现有明显相关性.     

前列腺癌患者外周血循环肿瘤细胞分型及自噬蛋白的表达对于远处转移的诊断价值

目的远处转移为前列腺癌的主要死亡原因,如何识别高转移风险的前列腺癌具有重要临床意义。本研究旨在通过CanPatrol技术检测前列腺癌外周血循环细胞(CTCs),并探究CTCs上皮间质转化(EMT)情况以及自噬相关蛋白MAP1S表达情况与前列腺癌远处转移等临床病理特征的关系。方法纳入2017年11月至2018年11月中山大学孙逸仙纪念医院泌尿外科确诊且未经治疗的前列腺癌患者。用CanPatrol系统检测外周血循环肿瘤细胞并根据上皮及间质标记物表达情况以及MAP1S表达情况进行分型,分析CTCs分型与前列腺癌临床病理特征的关系。结果根据纳入及排除标准,共纳入前列腺癌病例26例,其中CTCs检出率100%,间质型CTCs检出率92.30%。MAP1S阳性率88.46%,MAP1S阳性CTCs个数3.50(5.25)。分析CTCs分型及临床病理特征的关系发现,MAP1S阳性率高者远处转移风险高,间质型CTCs比例高者远处转移风险高,Gleason评分更高。结论本研究采用一种较新的CTCs检测系统,将间质标志物纳入检测体系,并同时检测了自噬标志物MAP1S表达情况。发现高间质型CTCs比例与远处转移及高Gleason评分相关,高MAP1S表达率与远处转移相关。提示检测外周血循环肿瘤细胞的EMT标志物与自噬标志物或可协助前列腺癌转移的早期诊断。     

循环肿瘤细胞数量及分型检测在肾癌疗效评价中的作用

目的探讨循环肿瘤细胞(CTCs)数量及分型检测在肾癌疗效评价中的作用。方法选择2015年6月至2016年9月我院收治的47例肾癌患者,行根治性肾切除术10例,肾部分切除术37例。记录术前患者一般状况信息,术中肿瘤分级分期等信息。术前及术后每3个月采集患者外周血7.5mL,应用纳米膜过滤、RNA原位杂交法进行CTCs数量及分型检测,并行胸部、上腹部、双肾计算机断层扫描(CT)等检测评估肿瘤有无远处转移。术后监测患者CTCs变化趋势,按以下方案进行分组:①影像学发现转移进展的患者,按进行治疗(包括切除转移灶或靶向治疗)和未予治疗分组;②CTCs呈进展趋势的患者(不论影像学是否发现转移灶),按进行分子靶向治疗和未进行靶向治疗分组。分别比较CTCs数量及间质型CTCs比例在2组患者中的差异。结果入组47例患者在随访期有5例发生病灶转移,其中针对2例个案(1例肺转移和1例双侧肾上腺转移患者)的CTCs实时监测发现,CTCs变化趋势与临床治疗转归一致,且均早于影像学检查发现疾病变化趋势。5例转移患者,3例给予治疗(1例行肾上腺转移灶切除,2例行分子靶向治疗),治疗后CTCs数量及间质型CTCs比例较治疗前均呈下降趋势,2例因患者原因未接受治疗,CTCs数量及间质型CTCs比例均呈进展趋势。CTCs呈进展趋势的患者16例,5例给予索拉菲尼分子靶向治疗(治疗2例,辅助治疗3例),11例未予治疗,靶向治疗组与未治疗组患者比较,CTCs数量及间质型CTCs比例下降。结论①CTCs具有评价肾癌治疗效果的作用,CTCs数量及间质型细胞比例的下降与治疗效果相关;②CTCs分型检测具有早期、灵敏的优势,可以作为实时监测治疗效果及疾病进展的手段;③分子靶向治疗具有降低肾癌CTCs数量及间质型CTCs比例的作用。     

肾细胞癌患者血液循环肿瘤细胞测定及临床意义

目的:采用α-FR的配体探针PCR定量检测肾细胞癌(RCC)患者外周血循环肿瘤细胞(CTCs),并阐释该法测定RCC患者外周血CTCs的初步临床意义。方法:体外ACHN掺血回收实验评估α-FR的配体探针PCR技术检测外周血CTCs的有效性。将2017年2月～2019年10月在我院就诊的50例健康对照者、35例肾脏良性病变者及45例初诊RCC患者纳入本临床试验;RCC患者分为术前、术后两组。采集入组者外周静脉血4 mL(RCC患者于术前1 d、术后7 d采血2次),用上述方法定量检测CTCs。结果:ACHN掺血回收实验提示本方法的检测值同掺入已知数量肿瘤细胞具有很高的相关性(r=0.99),RCC术前组外周血CTCs水平显著高于健康对照组和肾脏良性病变组(P0.01,P0.01),并且RCC患者CTCs水平同肿瘤的TNM分期、临床分期显著相关(P0.01),根治性肾切除术和保留肾单位手术对于T_1、T_2期不伴转移RCC患者术后外周血CTCs水平的影响无差异(P=0.320)。结论:采用α-FR的配体探针PCR能有效检测RCC患者外周血CTCs,本方法定量检测CTCs可弥补肾癌TNM分期的不足,可能在评估RCC患者治疗效果和监测肿瘤进展方面具有一定的价值。     

肾细胞癌VEGF表达和MVD检测的临床意义

目的 探讨肾细胞癌（RCC）组织中血管内皮细胞生长因子（VEGF）的表达与肿瘤间质微血管密度（MVD）检测的临床意义。方法 采用免疫组化方法对65例肾细胞癌（RCC）及10例正常肾组织进行VEGF单克隆抗体及CD34单克隆抗体染色，观察RCC的VEGF表达与MVD之间的相关性。结果 VEGF的表达与肿瘤间质微血管密度之间存在正相关性，二者均与RCC的病理分级、临床分期及远处转移显著相关（P〈0.05，P〈O．001）。结论 VEGF与MVD可客观准确反映RCC的生物学行为，上述二项指标可作为评估RCC恶性程度、转移及预后的重要指标。     

结肠癌患者循环肿瘤细胞与临床病理分期的相关性研究

目的了解结肠癌患者循环肿瘤细胞(CTCs)与临床病理分期之间的相关性。方法应用Cell Search技术检测未经治疗的结肠癌患者外周血中CTCs数量,并观察与临床病理分期的相关性。结果在32例首诊为结肠癌患者中,术前外周血检出CTCs阳性13例,阳性率40.6%;CTCs阳性率与患者年龄、性别、肿瘤大小、神经侵犯无关;与结肠癌浸润深度、TNM分期、脉管侵犯相关(P<0.05),TNM分期高(Ⅲ+Ⅳ)患者的CTCs阳性率显著高于TNM分期低(Ⅰ+Ⅱ)患者(P<0.05)。结论结肠癌患者外周血中检测到CTCs,提示其临床病理分期也可能较差。     

胃癌患者循环肿瘤细胞检测的临床意义

探讨胃癌患者检测循环肿瘤细胞（CTCs）的临床意义。选取2018年1月至2019年1月滨州医学院附属淄博市中心医院胃肠外科收治的胃癌患者60例（观察组）和同期26例良性胃病患者（对照组）作为研究对象。检测两组患者治疗前CTCs水平，同时检测胃癌组患者肿瘤标志物CEA水平。胃癌组患者CTCs阳性率为68.33%(41/60)，明显高于胃良性疾病患者组（7.69%,2/26），差异具有统计学意义（P<0.05）；胃癌组CTCs水平与患者年龄、性别、肿瘤直径大小、分化程度、淋巴结转移无关（P>0.05）；与肿瘤TNM分期、脉管侵犯、远处转移密切相关（P<0.05）。CTCs对胃癌的诊断阳性率为63.33%(38/60),CEA对胃癌的诊断阳性率为31.67%(19/60),CTC联合CEA对胃癌的诊断阳性率为73.33%(44/60)。胃癌患者CTCs水平显著升高；CTCs水平与胃癌临床分期、脉管癌栓、远处转移相关；CTCs联合CEA检测可以提高早期胃癌的阳性率。     

胃癌患者外周血循环肿瘤细胞检测及其临床意义

目的观察胃癌患者外周血循环肿瘤细胞(CTCs)的检出情况,探讨其与胃癌临床病理特征的关系。方法选取2011年9月至2013年9月期间在笔者所在医院经病理学检查证实的60例胃癌患者为研究对象,并与同期40例胃良性疾病对照;采集其静脉血,经Cell Tracks Auto Prep系统检测其CTCs阳性率,并分析CTCs与胃癌临床病理特征的关系。结果胃癌组CTCs检出率为70.0%(42/60),对照组CTCs检出率为7.5%(3/40),胃癌患者外周血组CTCs检出率显著高于胃良性疾病者(P0.05)。胃癌患者外周血CTCs检出率与患者性别、年龄、N分期、远处转移、肿瘤大小及脉管侵犯均无关(P0.05),而与肿瘤TNM分期及分化程度有关(P0.05)。外周血CTCs检测阴性的胃癌患者,其术后12个月及18个月的累积生存率均高于CTCs检测阳性者(P0.05)。结论 CTCs检测方便,胃癌患者外周血中CTCs检出率较高,其外周血CTCs的检出情况可以反应胃癌的进展程度,可作为其预后判断的指标。     

循环肿瘤细胞与胃癌患者临床病理特征及预后的关系研究

目的检测胃癌患者空腹外周静脉血中循环肿瘤细胞(CTCs)水平并探讨其与胃癌患者临床病理特征和预后的关系。方法选取就诊于中国人民解放军联勤保障部队第九四〇医院2015年8月至2016年12月期间经组织病理学证实为胃癌且行胃癌根治术的患者100例(胃癌组),选取同时期就诊于笔者所在医院的胃良性病变患者38例作为对照(胃良性病变组)。抽取2组患者清晨空腹肘正中静脉血7 mL,24 h内采用免疫磁微粒阴性富集法联合免疫荧光原位杂交技术检测CTCs,计算CTCs阳性率并分析其与胃癌患者临床病理特征(肿瘤部位、浸润深度、分化程度、TNM分期、淋巴结转移及有无脉管癌栓)及无进展生存期的关系。结果胃癌组患者外周静脉血中CTCs阳性率为89.0%(89/100),明显高于胃良性病变组的10.5%(4/38),2组间比较差异有统计学意义(P0.001)。胃癌患者外周静脉血中CTCs水平与肿瘤浸润深度(P=0.017)、淋巴结转移(P=0.038)及TNM分期(P=0.016)有关,而与患者性别、年龄、肿瘤部位、分化程度及有无脉管癌栓无关(P0.050)。CTCs对胃癌诊断的预测价值明显优于肿瘤标志物CEA、CA19-9和CA125(受试者操作特征曲线下面积0.935比0.711、0.666、0.551)。胃癌患者中CTCs低表达组术后无进展生存期明显长于高表达组(χ~2=5.172,P=0.023)。结论免疫磁微粒阴性富集法联合免疫荧光原位杂交技术检测患者空腹外周静脉血中CTCs水平对胃癌诊断具有较高的敏感度和诊断价值,其对指导临床肿瘤分期及对胃癌患者预后的预测也有一定的价值。     

NF-κBp65蛋白和自噬相关因子Beclin1及p62在甲状腺癌中的表达及临床意义

目的分析核因子NF-κB(NF-κB)p65和自噬相关蛋白Beclin1及p62在甲腺乳头状癌(PTC)中的表达及其临床意义。方法收集2013年3月至2015年2月期间笔者所在医院收治的160例经病理学检查证实为PTC患者的肿瘤组织标本及其癌旁组织标本,同时收集上述患者中伴有颈部淋巴结转移组织标本90例。采用免疫组化方法检测PTC组织、癌旁组织和转移淋巴结组织中NF-κBp65、Beclin1及p62的表达情况,分析上述指标与PTC患者临床病理特征及预后的关系。结果 NF-κBp65和p62在PTC组织和转移淋巴结组织中的表达阳性率均高于癌旁组织(P0.05),而Beclin1在PTC组织和转移淋巴结组织中的表达阳性率均低于癌旁组织(P0.05)。PTC组织中NF-κBp65的表达与患者的临床病理特征均无关(P0.05);p62的表达随肿瘤分化程度升高而降低(P0.05);Ⅲ+Ⅳ期和有淋巴结转移患者中的Beclin1表达分别低于Ⅰ+Ⅱ期和无淋巴转移者(P0.05),而p62的表达与之相反。Spearman相关分析显示,PTC组织中,Beclin1和p62的表达呈负相关关系(r=–0.656,P0.01),在转移淋巴结组织中,Beclin1和p62的表达也呈负相关关系(r=–0.562,P0.01)。PTC组织中p62及NF-κBp65表达阳性者3年生存率均低于表达阴性者(P0.05),Beclin1表达阳性者3年生存率则高于表达阴性者(P0.05)。TNM分期、淋巴结转移、NF-κBp65及p62为PTC预后的独立危险因素,而Beclin1为保护因素。结论NF-κBp65和p62在PTC组织及淋巴结转移组织中呈高表达,而Beclin1呈低表达,三者可作为PTC患者预后独立预测因素;Beclin1及p62与PTC生物学行为有关,可能成为PTC诊断的潜在指标。     

Prognostic value of circulating tumor cells and immune-inflammatory cells in patients with renal cell carcinoma

PurposeThe relationships among circulating tumor cells (CTCs), inflammatory cells, and platelets in patients with renal cell carcinoma (RCC) are not transparent. We evaluated the correlations among CTCs, blood inflammatory cells, and platelets in patients with RCC and their prognostic value for metastasis-free survival.MethodsCTC and typical tumor cell chip data were collected and analyzed by the GEO database. The baseline data, survival data, CTCs data, and blood test results were statistically analyzed.ResultsBioinformatics analysis showed that the function of the differentially expressed genes between CTCs and normal tumor cells mainly involved platelets and immune inflammation. A total of 82 patients whose follow-up time was 3 to 68 months were included in the analysis. Clinical data of the patients confirmed that there is a correlation between platelets and mesenchymal CTCs. Simultaneously, there was a correlation between immune inflammatory cells and platelets. The univariate Cox proportional hazards model indicated that staging, mesenchymal CTCs, and the monocyte-to-neutrophil ratio (MNR) had prognostic value. The multivariate Cox proportional hazards model indicated that staging and the MNR had prognostic value and high accuracy.ConclusionsBioinformatics analysis showed that CTCs were related to platelets and immune-inflammatory cells. Furthermore, the clinical data confirmed that platelets were correlated with mesenchymal CTCs and immune-inflammatory cells in the blood. By using mesenchymal CTCs, the MNR, or staging respectively, it is possible to predict the risk of postoperative metastasis in RCC patients. As a compound prognostic factor, staging, and the MNR can provide more convenient and accurate condition monitoring.     

A preliminary study on the association between epithelial-mesenchymal transition and circulating tumor cells in renal cell carcinoma

BackgroundCirculating tumor cells (CTCs) are considered useful prognostic factors for various cancers, and in 2014, our research group conducted a comparative experiment of CTC detection in patients with renal cell cancer (RCC). However, the reason for the low detection rate of CTCs in cancer patients using the CellSearch^® system is still unknown, although it has been hypothesized to be attributed to the likelihood that CTCs undergoing epithelial-mesenchymal transition (EMT) do not express the CTC biomarkers cytokeratin (CK)8/18/19 or epithelial cell adhesion molecule (EpCAM). The overall aim of the current study was to investigate the expression levels of CK8/18/19 and EpCAM in relation to the EMT biomarkers vimentin and E-cadherin in patients with RCC.MethodsPatients with RCC who had undergone radical nephrectomy or partial resection between May 2014 and December 2014 were initially recruited.ResultsAmong 34 RCC patients, nine co-expressed EpCAM and CK8/18/19 in primary tumor tissues. The CellSearch^® results showed that CK8/18/19 was expressed in 5 of 6 patients (5/6) and EpCAM was expressed in 6 patients (6/6). However, the isolation by size of tumor cells (ISET) technique showed these were co-expressed in only four of the 10. The expression of CK8/18/19, EpCAM, vimentin, and E-cadherin was distributed unequally in different enumeration groups of CTCs (all P>0.05), and the positive expression of CK8/18/19 was correlated with neutrophil number and tumor size (P<0.05). The positive expression of vimentin was correlated with the Karnofsky Performance Status (KPS) score and clinical stage of renal cancer patients (P<0.05).ConclusionsOur results indirectly proved the occurrence of EMT in the formation of CTCs by comparing and analyzing the expression of CK8/18/19 and EpCAM in renal cancer tissues and the detection results of CTCs.     

Comparison of two detection systems for circulating tumor cells among patients with renal cell carcinoma

Background/aims

Detection of circulating tumor cells (CTCs) in cancer patients has diagnostic and prognostic importance. However, the clinical implications of CTC detection in patients with renal cell carcinoma (RCC) are still unclear. In this study, we investigated the clinical significance of CTCs using two detection systems, the CellSearch system (CSS) and isolation by size of epithelial tumor cells (ISET), among RCC patients.

Methods

We recruited 36 RCC patients and 22 healthy volunteers as controls. Blood was drawn before treatment. Samples were analyzed using the CSS and ISET. We prospectively followed the RCC patients to determine overall and progression-free survival.

Results

We did not detect CTCs in the control group using either the CSS or ISET. CTCs were detected in 7/36 patients (19.4%) using the CSS and in 13/36 patients (36.1%) using ISET, while circulating microemboli (CTMs) were detected in three patients (8.3%). The presence of ISET-detected CTCs correlated with clinical tumor node metastasis (TNM) stages, while the CSS-detected CTCs did not. After 36 months (median), CTCs detected by both methods failed to correlate with overall and progression-free survival among RCC patients.

Conclusion

We discovered that ISET is more suitable than the CSS for detecting CTCs in RCC patients. The presence of CTCs/CTMs in RCC patients correlated with higher TNM stages, suggesting that the presence of CTCs could be a prognostic marker in RCC patients.

     

Correlation Between Postoperative Early Recurrence of Hepatocellular Carcinoma and Mesenchymal Circulating Tumor Cells in Peripheral Blood

Background

Circulating tumor cells (CTCs) have been actively studied for their functions in hepatocellular carcinoma (HCC) recurrence. However, the relationship between circulating tumor cells subtypes and hepatocellular carcinoma recurrence is still unclear.

Methods

CTCs were collected from the peripheral blood of 62 postoperative HCC patients. The CTCs were isolated with a filtration-based method. Multiplex fluorescence in situ hybridization was used to characterize the CTCs based on mRNA expression levels of epithelial and mesenchymal markers.

Results

Of the 62 HCC patients, 26 were diagnosed with early recurrence (ER) and 36 did not experience recurrence. Comparison between the recurrence group and the non-recurrence group showed the total number of CTCs, mesenchymal CTCs, and mixed CTCs in the recurrence group was significantly higher than in the non-recurrence group. Receiver operator characteristic (ROC) curve analysis was performed to define the positive cutoff values as follows: total number of CTCs ≥?4, mesenchymal CTCs ≥?1, and mixed CTCs ≥?3. Analysis showed that portal vein tumor thrombus (hazard ratio [HR] = 2.905, P = 0.023) and mesenchymal CTC positivity (HR = 3.453, P = 0.007) were independent risk factors for ER. The correlation between the presence of mesenchymal CTCs and time to recurrence was further examined, and the results showed significantly shortened postoperative disease-free survival in patients positive for mesenchymal CTCs (P < 0.001).

Conclusions

HCC patients with positive peripheral mesenchymal CTCs have a more serious risk of ER, which could be a potential biomarker in HCC prognosis monitoring.

     

Relationship between Circulating Tumor Cells,Blood Coagulation,and Urokinase‐Plasminogen‐Activator System in Early Breast Cancer Patients

Cancer is a risk factor for venous thromboembolism (VTE) and plasma d‐dimer (DD) and tissue factor (TF) are established VTE associated markers. Circulating tumor cells (CTCs) are associated with the risk of VTE in metastatic breast cancer. This study aimed to correlate CTCs, blood coagulation and the urokinase plasminogen activator (uPA) system in primary breast cancer (PBC) patients. This prospective study included 116 PBC patients treated by primary surgery. CTCs were detected by quantitative RT‐PCR assay for expression of epithelial (CK19) or epithelial‐mesenchymal transition (EMT) genes (TWIST1, SNAIL1, SLUG, ZEB1, FOXC2). Plasma DD, TF, uPA system proteins were detected by enzyme‐linked immunosorbent assays, while expressions of uPA system in surgical specimens were evaluated by immunohistochemistry. CTCs were detected in 27.6% patients. Patients with CTCs had a significantly higher mean plasma DD (ng/mL) than those of patients without CTCs (632.4 versus 365.4, p = 0.000004). There was no association between plasma TF and CTCs. Epithelial CTCs exhibit higher expression of uPA system genes compared to EMT_CTCs. Patients with CTCs had higher plasma uPA proteins than those of patients without CTCs; there was no correlation between tissue expression of uPA system, CTCs, DD or TF levels. In multivariate analysis CTCs and patients age were independent factors associated with plasma DD. We found association between plasma DD and CTCs indicating a potential role for activation of the coagulation cascade in the early metastatic process. CTCs could be directly involved in coagulation activation or increased CTCs could be marker of aggressive disease and increased VTE risk.     

p16蛋白及PCNA在肾癌中的表达及意义

目的探讨肾癌中p16蛋白与PCNA的表达与肾癌组织学类型、临床分期、病理分级及预后的关系。方法采用免疫组织化学SP法检测41例肾癌及18例癌旁正常肾组织中p16蛋白及PC－NA的表达。结果p16蛋白在肾癌及癌旁正常肾组织中的阳性表达率间具有显著性差异（P＜0.05）,p16蛋白表达在肾透明细胞癌及颗粒细胞癌间无显著性差异,随肾癌分级、分期升高而明显降低,p16阳性表达组术后5年生存率明显高于阴性表达组;肾癌及癌旁肾组织中PCNA的阳性表达率增高,与预后有关,与肾癌的组织类型及分期无关。结论p16基因及蛋白的检测可作为肾癌的辅助诊断及预后判断的参考指标;PCNA可作为肿瘤恶性程度的判断及预后的参考指标。     

Molecular characterization of circulating tumor cells in pancreatic ductal adenocarcinoma: potential diagnostic and prognostic significance in clinical practice

BackgroundThe clinical value of heterogeneous sub-populations of circulating tumor cells (CTCs) in pancreatic ductal adenocarcinoma (PDAC) remains unclear.MethodsPeripheral blood samples were obtained from 67 PDAC patients. CTCs were isolated by employing CD45 negative enrichment technique and further characterized for epithelial to mesenchymal transition (EMT) or human equilibrative nucleoside transporter-1 (hENT-1). The relationships between CTCs sub-phenotypes with clinicopathological factors or post-operative recurrence in PDAC patients were analyzed.ResultsEMT related CTCs could be isolated and identified from the 81% of patients (54/67), and both the total count (median: 5 vs. 17/mL, P<0.0001) and M-CTC percentage (median: 0.2 vs. 0.345, P=0.0244) of CTCs could differentiate local/regional with metastatic disease. Multivariate analysis showed that both AJCC stage (P=0.025) and M-CTC percentage (P=0.001) were independent prognostic indicators of recurrence free survival (RFS) in resected patients. Moreover, Kaplan-Meier survival analysis showed that M-CTC after 2 courses of chemotherapy was significantly associated with inferior RFS (49.5 weeks vs. undefined, P=0.0288). No significant correlation in hENT-1 expression was found between CTCs and matched tumor tissues, and further multivariate analysis suggested hENT-1 expression in CTCs as independent prognostic factor for RFS (P=0.016). Patients with low hENT-1 expression in CTCs had decreased RFS (32 weeks vs. undefined, P=0.0337).ConclusionsCTCs could be the promising diagnostic biomarkers in PDAC patients, and phenotypic profiling of CTCs based on EMT or hENT-1 could help establish novel prognostic biomarkers in resected patients undergoing adjuvant gemcitabine-based chemotherapy.KeywordsCirculating tumor cells (CTCs); Pancreatic ductal adenocarcinoma (PDAC); Epithelial to mesenchymal transition (EMT); human equilibrative nucleoside transporter-1 (hENT-1)     

P-gp和MRP在肾癌中的表达及其临床意义

目的　探讨 p -糖蛋白(P- gp)和多药耐药相关蛋白(MRP)在肾癌中表达的临床意义及相互关系。方法　采用免疫组织化学方法对106例肾癌石蜡包埋标本进行P- gp和 MRP检测。结果　P- gp及 MRP在肾癌中的阳性表达率分别为68.8%(73/106)和 56.6%(60/106)。肾癌P- gp的表达与性别、病理类型、临床分期及预后无关,但与病理分级有关。肾癌 MRP的表达与性别、病理类型无关,但MRP的表达随肾癌病理分级、临床分期的升高而增强。MRP阳性表达组 3 年生存率显著低于 MRP阴性表达组。P- gp与MRP的表达具有相关性。结论　P- gp和 MRP过表达对肾癌固有的多药耐药现象可能具有一定的协同作用。MRP参与了肾癌的恶性进展并可作为肾癌生物学行为及预后评估的参考指标。     

间隙连接蛋白43在肾细胞癌中的表达及意义

目的:探讨间隙连接蛋白Cx43在肾细胞癌(RCC)中的表达及其与RCC生物学行为之间的关系.方法:应用S-P免疫组织化学法检测Cx43蛋白在41例RCC,12例癌旁肾组织及10例正常肾组织中的表达情况.结果:Cx43阳性染色主要定位在细胞膜和细胞质上.Cx43蛋白在RCC中的阳性表达率明显低于在癌旁肾及正常肾组织的水平(P＜ 0.01);在透明细胞癌、颗粒细胞癌、梭形细胞癌中,其阳性表达率比较差异无统计学意义(P＞ 0.05);随着临床分期的增高,其阳性率明显下降(P＜ 0.05),并与RCC转移呈负相关(Spearman 等级相关系数r=- 0.483, P＜ 0.01);与肿瘤大小无明显关系(P＞ 0.05).结论:Cx43对RCC发生和转移有明显抑制作用,其表达减弱或消失可能与RCC的发生和发展密切相关.