乳腺癌患者外周血表皮生长因子受体mRNA与组织中蛋白表达的相关性及其意义

 目的 本研究旨在检测乳腺癌患者外周血EGFRmRNA的表达，评价其作为循环乳腺癌细胞标志的可能性；结合相应肿瘤组织中EGFR蛋白的检测，分析两者的相关性；并结合临床病理资料综合分析，探讨其在乳腺癌患者外周血中表达的意义。方法 以RT-PCR法，分别对40例乳腺癌患者、25例良性乳腺病患者和30例健康人外用静脉血进行EGFRmRNA检测。免疫组化法检测相应肿瘤组织中EGFR蛋白的表达情况。结果 乳腺癌组外周静脉血中EGFRmRNA检出率（32．5％）与健康人组（0）和良性乳腺病患者组（0）间差异均存在统计学意义（P〈0．01）。免疫组化法检测到乳腺癌患者相应肿瘤组织中有25例（62．5％）EGFR蛋白阳性，组织中EGFR蛋白表达级别高的患者其相应外周血EG—FRmRNA的检出率也高。EGFRmRNA阳性表达与临床分期、组织学分级及ER闰存在明显的相关性，而与病理类型、肿瘤大小、腋窝淋巴结转移、PR和HER-2间无明显相关性。结论 RT-PCR法检测乳腺癌患者外周血EGFRmRNA有望成为检测循环乳腺癌细胞的有效方法。结合临床病理资料分析，RT—PCR法检测外周血EGFRmRNA有助于判断乳腺癌的预后，有益于乳腺癌微转移的早期诊断、治疗方案的选择及治疗疗效的监测。     

外周血CK-19mRNA检测对乳腺癌预后的影响

目的:探讨外周血CK-19mRNA对乳腺癌患者预后的影响.方法:对86例初治乳腺癌患者根治手术后的外周血用RT-PCR方法检测了CK-19mRNA的表达,并采用S-P免疫组化法检测了手术标本的c-erbB-2、ER、PR蛋白的表达情况.结果:手术后患者外周血CK-19mRNA阳性率为52.3%;外周血CK-19mRNA状况与临床分期呈显著相关(P＜0.05),而与肿瘤大小、腋淋巴结转移、c-erbB-2、ER、PR蛋白的表达均无显著相关(P＞0.05);外周血CK-19mRNA阴性病例与阳性病例相比,无病生存期和总生存期显著延长(P＜0.05).结论:乳腺癌患者术后外周血CK-19mRNA的检测有助于评估预后.     

Detection of circulating breast tumor cells by differential expression of marker genes.

PURPOSE: We undertook a systematic approach to identify breast cancer (BC) marker genes with molecular assays and evaluated these marker genes for the detection of minimal residual disease in peripheral blood mononuclear cells (PBMCs). EXPERIMENTAL DESIGN: We used serial analysis of gene expression to identify a range of genes that were expressed in BC but absent in the expression profiles of blood and bone marrow cells. Next, we evaluated a panel of four marker genes (p1B, PS2, CK19, and EGP2) by real-time quantitative PCR in 103 PBMC samples from patients with metastatic BC (stage III/IV) and in 96 PBMC samples from healthy females. RESULTS: Increased marker gene expression of at least one marker was seen in 33 of 103 patients. Using quadratic discriminant analysis including all four marker genes, we determined a discriminant value with 29% positivity in the BC patient group that did not yield false positive results among the healthy females. CONCLUSIONS: Real-time PCR for the simultaneous expression of multiple cancer-specific genes may ensure the specificity required for the clinical application of mRNA expression-based assays for occult tumor cells.     

HMGA2 is associated with invasiveness but not a suitable marker for the detection of circulating tumor cells in breast cancer

Previously, the human high mobility group protein member HMGA2 mRNA was reported to be expressed in peripheral blood of patients with breast cancer, but not in healthy individuals. Expression of HMGA2 in blood was suggested to be an independent indicator of poor prognosis in metastatic breast cancer. These very promising findings propose HMGA2 as a potential marker for the detection of circulating tumor cells in peripheral blood. Therefore, we analyzed peripheral blood specimens from healthy controls and patients with breast tumors for HMGA2 expression using TaqMan real-time RT-PCR to test if HMGA2 is a suitable marker for the early detection of breast cancer and monitoring therapy response in peripheral blood. Furthermore, we examined the possible involvement of HMGA2 expression in invasion investigated by an in vitro invasion assay using established breast cell lines. HMGA2 expression was detected in peripheral blood of breast cancer patients as well as of healthy individuals. No significant association of HMGA2 expression with any clinical or histopathological data was apparent. However, there was a significant correlation of HMGA2 expression in invasive and non invasive breast cell lines (p=0.0056). Although, HMGA2 obviously contributes to invasion it is not a specific marker for the detection of circulating tumor cells in peripheral blood.     

肝癌患者围手术期外周血甲胎蛋白mRNA表达与术后复发的关系

背景与目的:甲胎蛋白(alpha-fetoprotein,AFP)mRNA是检测肝癌患者外周血中癌细胞常用的标志物。本研究探讨肝癌患者围手术期的外周血中AFPmRNA表达与术后复发的关系。方法:应用巢式PCR和Taq Man MGB探针法PCR定量技术,检测56例肝细胞癌患者术前和术后外周血、15例良性肝占位性病变合并肝硬化患者外周血以及30例健康志愿者外周血AFP mRNA的表达情况。结果:肝癌患者术前外周血AFP mRNA的阳性率为42.9%(24/56),高于良性肝占位合并肝硬化患者的13.3%(2/15)和健康对照的10.0%(3/30),其差异均有统计学意义(P=0.035,P=0.002)。术前AFP mRNA的表达与肿瘤有无肉眼及镜下血管侵犯的相关性有统计学意义(P=0.029,P<0.001)。AFP mRNA阴性患者1、2、3年生存率及无瘤生存率均比AFP mRNA阳性患者术后高(P=0.003,P=0.039)。Cox多因素分析结果显示术前外周血AFP mRNA阳性是预测术后复发的独立因素(P=0.018)。术后患者外周血AFP mRNA的阳性率为37.5%(21/56),术后AFP mRNA的表达与患者各项临床病理特征、预后以及术前AFP mRNA水平的相关性均无统计学意义。结论:肝癌患者术前外周血中AFP mRNA阳性者比AFP mRNA阳性者肿瘤侵袭性更强,术后可能更易复发。     

ECT2 mRNA及蛋白在胃癌患者外周血中的表达及意义

目的:探讨ECT2 mRNA及蛋白在胃癌患者外周血中的表达水平及其意义。方法:应用逆转录聚合酶链式反应（RT-PCR）及酶联免疫吸附实验（ELISA）测定50例胃癌患者及30例健康志愿者外周血ECT2 mRNA及蛋白的表达水平。结果:胃癌患者外周血中ECT2 mRNA阳性表达率为54%,健康志愿者外周血中ECT2 mRNA均阴性表达(P<0.05)。ECT2蛋白在胃癌患者外周血中的浓度（1 306.389±215.824)ng/L显著高于健康志愿者外周血中的浓度（502.718±69.440)ng/L(P<0.05),并且与TNM分期、淋巴结转移和浸润深度相关(P<0.05)。结论:外周血中ECT2 mRNA及蛋白的表达与胃癌关系密切,可作为反映胃癌发生、发展过程的有效分子指标。     

人乳腺癌外周血中hMAM mRNA表达及临床意义

目的：研究人乳腺癌外周血珠蛋白（hMAM）mRNA表达及临床意义。方法：应用nested-RT-PCR技术检测50例乳腺癌外周血hMAM mRNA的表达和化疗前后外周血hMAM mRNA的变化,并取20例乳腺纤维腺瘤、10例健康志愿者作阴性对照。结果：50例乳腺癌外周血hMAM mRNA的阳性表达率为34.0%;20例乳腺纤维腺瘤及10例健康志愿者外周血中均未检出hMAM mRNA的表达。hMAM mRNA的阳性与淋巴结转移状况、肿瘤TNM分期之间差异有统计学意义;50例乳腺癌化疗前检测17例hMAM mRNA阳性,化疗2～3周期后13例hMAM mRNA转为阴性,转阴率76.5%（13/17）,化疗前后差异非常显著（P=0.001）。结论：乳腺癌外周血中hMAM mRNA表达可以反映术后化疗对乳腺癌血液微转移的影响,有望成为乳腺癌血道微转移标志物和一个重要的预后指标。     

c-erbB-2 protein level in tissue and sera of breast cancer patients: a possibly useful clinical correlation

AIMS AND BACKGROUND: The aims of this study were to assess the clinical utility of circulating preoperative HER-2 extracellular domain p105 detected by enzyme immunoassay (ELISA), to compare the tissue expression of HER-2/neu determined by immunohistochemistry (IHC), to correlate prognostic factors including tumor size, nodal involvement, and hormone receptor status, and to analyze the prognostic significance of the marker in relation to clinical outcome as measured by disease-free and overall survival. METHODS: In this study, we enrolled 108 consecutive patients with breast carcinoma, and obtained serum samples and frozen tumor tissues. We compared them with 57 women with fibroadenoma and 63 healthy women as controls. RESULTS: Univariate ANOVA analysis showed no relationship between HER-2/neu in tissue and serum. Preoperative serum levels of p105 were significantly higher in breast cancer patients than in women with benign disease or healthy women. Concerning the correlation between p105, HER-2/neu tissue expression, and the other prognostic factors, a statistically significant correlation between high serum p105 levels and ER-negative status in breast cancer patients was found. Kaplan-Meier analysis confirmed that patients with positive HER-2/neu tissue expression had a significantly shorter survival than those with negative expression. Analysis with the Cox model demonstrated that tumor size was the only significant independent prognostic factor. CONCLUSIONS: This research failed to demonstrate a relationship between preoperative tissue overexpression and circulating HER-2/neu, suggesting that p105 does not represent a valid alternative to predict a worsened prognosis in breast cancer, but it could be a diagnostic marker to discriminate healthy subjects from breast cancer patients.     

非小细胞肺癌患者外周血CD44V6 mRNA表达与微转移的相关性研究

目的 探讨CD44V6 mRNA表达与非小细胞肺癌(NSCLC)外周血微转移的关系及临床意义,及其与CK19 mRNA在外周血中联合检测的临床意义.方法 应用免疫组化SP法检测56例NSCLC癌组织中CD44V6 mRNA的表达情况,应用逆转录聚合酶链反应(RT-PCR)技术检测外周血中CD44V6 mRNA与CK19 mRNA的表达情况.采集20例肺部良性病变患者正常肺组织和外周血作对照.结果 CD44V6 mRNA在NSCLC癌组织中表达明显高于良性病变正常肺组织(P<0.001),其阳性表达率在各病理分期间、有无淋巴结转移组间的差异具有统计学意义(P<0.05),而在各病理类型间差异无统计学意义(P>0.05).NSCLC患者外周血中CD44V6 mRNA阳性表达率高于对照组(P<0.05),其阳性表达率在各病理分期间、有无淋巴结转移间的差异具有统计学意义(P<0.05).CD44V6 mRNA在NSCLC组织中表达与外周血中表达呈正相关,NSCLC外周血中CD44V6 mRNA与CK19 mRNA表达呈正相关;CD44V6 mRNA与CK19 mRNA联合检测灵敏度为75.0%,优于单基因检测(P<0.05).结论 NSCLC外周血中CD44V6 mRNA的高表达与其侵袭转移有关,CD44V6 mRNA可作为检测NSCLC外周血微转移的分子肿瘤标志物,并有望成为判定NSCLC预后的分子标志物;CD44V6 mRNA与CK19 mRNA联合检测可提高NSCLC血行微转移诊断的敏感度.     

外周血人类斯钙素-1基因表达与乳腺癌微转移的关系

目的:检测乳腺癌患者外周血、肿瘤组织中hSTC-1mRNA的表达,判断外周血微转移状况,探讨微转移与肿瘤恶性程度之间的关系。方法:采用RT-PCR法检测乳腺癌组织、外周血中hSTC-1的mRNA表达,用免疫组化法检测乳腺肿瘤组织中ER、PR及VEGF的表达。结果:hSTC-1mRNA在乳腺癌患者癌组织中的阳性率为93.3%(28/30),在外周血中的阳性率为37.3%(19/51);17例非肿瘤成年女性患者外周血中未检测到hSTC-1的mRNA。血液中的hSTC-1的mRNA表达与多个临床病理因子有相关性(P<0.05),包括:肿瘤大小、腋淋巴结转移、TNM分期、ER、VEGF等;结论:乳腺癌患者外周血检测到hSTC-1基因的表达,说明乳腺癌已发生血行微转移,乳腺癌患者外周血hSTC-1的mRNA表达与多个临床病理因子有关;这些因子决定乳腺癌的预后,因此hSTC-1可以作为检测乳腺癌外周血微转移的一种新的分子标记。     

Overexpression of beta 1,4N-acetylgalactosaminyl- transferase mRNA as a molecular marker for various types of cancers

OBJECTIVE: To determine GalNAcT mRNA expression in human carcinoma cell lines and primary tumor tissues. Assessment of the potential use of GalNAcT mRNA as a molecular marker for detection of metastatic cancer cells in the peripheral blood of patients with hepatocellular carcinoma. METHODS/RESULTS: We investigated GalNAcT mRNA expression in various human cancer cell lines and primary cancer tissues using RT-PCR assay for GalNAcT mRNA. The expression of GalNAcT mRNA was detected in 25 of 26 cancer cell lines tested and in the majority of primary tumors from different organs: 8 of 10 colon cancers, 9 of 9 breast cancers, 11 of 12 esophageal cancers, 14 of 14 gastric cancers, 4 of 18 pancreatic cancers, 6 of 12 biliary tract cancers, 17 of 18 hepatocellular carcinomas and 13 of 14 lung cancers. Semi-quantitative analysis with duplex RT-PCR showed that the amount of the GalNAcT mRNA was enhanced in cancer tissues as compared to the surrounding cancer-free tissues. Blood specimens of 5 of 14 patients with hepatocellular carcinoma were positive for GalNAcT mRNA, all of whom developed recurrent disease in less than 24 months. Peripheral blood samples of 30 normal subjects were negative for GalNAcT mRNA. CONCLUSION: Our results suggest that the RT-PCR assay for GalNAcT mRNA could be a potentially useful molecular marker for detecting cancer dissemination in blood circulation of patients with malignancy.     

Detection of HER-2/neu-positive circulating epithelial cells in prostate cancer patients

HER-2/neu may play a role in prostate carcinogenesis. The aim of this study was to use the expression of HER-2/neu as a molecular marker for the detection of circulating tumour cells (CTCs) in the blood of patients with prostate cancer (PC). Blood samples were collected from 42 patients with PC and nine healthy volunteers. Immunomagnetic beads were used to harvest epithelial cells from peripheral blood mononuclear cells. Total RNA was extracted and reverse transcribed before analysis by real-time PCR with HER-2/neu-specific primers. CTCs were HER-2/neu positive in six out of 11 (54%) patients with metastatic disease and in three out of 31 (9.6%) patients with localised PC (P=0.004). In blood samples from nine healthy volunteers, we detected no expression of HER-2/neu. The present method appears to be minimally invasive, highly sensitive and a specific approach for detecting CTCs in PC. Furthermore, it may help better target HER-2/neu in advanced PC.