Breast cancer micrometastases: Different interactions of carcinoma cells with normal and cancer patients' bone marrow stromata

The apparently dormant breast cancer micrometastases in haemopoietic marrow are correlated with distant metastatic carcinoma dissemination. We studied in vitro interactions of carcinoma cells with adjacent stromata, using connective tissue cell cultures from breast and bone marrow samples of normal donors, comparing them to the pericancerous breast tissue and bone marrows of 12 selected patients with invasive breast carcinomas. Cancer cells were detected by immunocytochemistry and RT-PCR in all the bone marrows and in most blood samples of the studied patients. We monitored the growth and interaction of carcinoma MCF-7 cells with the stromata. The normal breast stroma sustained typical massive cancer growth. The pericancerous breast stroma induced the invasive mesenchymal pattern of growth. Normal bone marrow stroma induced the same conversion and was highly adhesive, retaining the cells in the stroma, but carcinoma patients' bone marrow stromata underwent low adhesive interactions with cancer cells, releasing them potentially into the circulation. The semi-quantitative RT-PCR indicated an enhanced expression of the hepatocyte growth factor and its receptor c-met in breast and bone marrow stromata of cancer patients. The input of cancer cells into the normal bone marrow may induce modifications of the local microenvironment, favourable for growth and release of carcinoma cells into the systemic circulation, which correlate with the poor prognosis of patients with bone marrow micrometastases. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chemokine receptor CXCR4 expression in breast cancer as a potential predictive marker of isolated tumor cells in bone marrow

Interactions between the CXCR4 chemokine receptor in breast cancer cells and the ligand CXCL12/SDF-1α are thought to play an important role in breast cancer metastases. In this pilot study, CXCR4 expression along with other biomarkers including HER2-neu and EGFR, were measured in primary tumor samples of patients with operable breast cancer to test whether any of these biomarkers alone and in combination could indicate breast cancer with high likelihood of metastasizing to bone marrow. Cytokeratin (CK) positive cells in bone marrow were identified by flow-cytometry following enrichment with CK 7/8 antibody-coupled magnetic beads. Primary tumors (n = 18) were stained with specific antibodies for CXCR4, HER2-neu, EGFR, and PCNA using an indirect avidin–biotin horseradish peroxidase method. The majority of the patients had T2/T3 tumors (72%), or lymph node involvement (67%) as pathologic characteristics that were more indicative of high-risk breast cancer. High CXCR4 cytoplasmic expression was found in 7 of 18 patients (39%), whereas 6 of 18 patients (33%) were found to have CK positivity in bone marrow. The median number of CK⁺ cells was 236 (range, 20–847) per 5 × 10⁴ enriched BM cells. The presence of CK⁺ cells in bone marrow was found to be associated with increased expression of CXCR4 alone or in addition to EGFR and/or HER2-neu expression (P = 0.013, P = 0.005, and P = 0.025, respectively) in primary tumors. Furthermore, three patients with high CK positivity (>236 CK⁺ per 5 × 10⁴ enriched bone marrow cells) in bone marrow exclusively expressed high levels of CXCR4 with EGFR/HER2-neu (P = 0.001). Our data suggest that high CXCR4 expression in breast cancer may be a potential marker in predicting isolated tumor cells in bone marrow. CXCR4 coexpression with EGFR/HER2-neu might further predict a particular subset of patients with high CK positivity in bone marrow.     

乳腺癌骨髓微转移状况与ER PR VEGF 及Cath-D等因子的关系

目的:检测乳腺癌骨髓组织CK19 mRNA的表达,判断骨髓微转移状况,探讨其与各因子的关系.方法:采用RT-PCR法(一步法)检测骨髓组织中CK19 mRNA表达;用免疫组化SP法检测乳腺癌组织中ER、PR、Cath-D、C-erbB-2及VEGF的表达.结果:102例乳腺癌患者骨髓组织CK19 mRNA表达阳性率56.9%.骨髓CK19 mRNA表达随乳腺癌组织中ER及PR蛋白表达增强而降低(P＜0.05,P＜0.01);随Cath-D、VEGF蛋白表达增强而呈增高趋势(P＜0.005,P＜0.05);但与C-erbB-2表达状况无关.结论:乳腺癌骨髓微转移与癌组织中某些生物学因子表达有关;这些因子决定癌细胞的生物学行为.     

非小细胞肺癌患者淋巴结、外周血、骨髓微转移的MUC1基因诊断及临床意义

目的 :探讨非小细胞肺癌 (NSCLC)患者淋巴结、外周血、骨髓微转移的MUC1基因诊断的临床意义 ,以及三者微转移之间的相关性。方法 :应用逆转录聚合酶链反应 (RT PCR)技术 ,联合检测 31例肺癌患者和 10例肺良性病变患者的淋巴结、外周血和骨髓中MUC1基因mRNA表达。结果 :本实验建立的巢式RT PCR技术的敏感性达 10 -6。术前 10例患者外周血、7例患者骨髓检测到肺癌微转移。手术取 119枚淋巴结中 6 5枚检测到肺癌微转移。肺癌患者淋巴结、外周血、骨髓微转移阳性检出率分别为 5 4 .6 %、32 .3%、2 2 .6 % ,三者之间存在正相关(P <0 .0 5 )。结论 :RT PCR法是一种特异性、敏感性均较高的肿瘤微转移检测方法 ;MUC1基因mRNA可能是检测肺癌微转移的一个有价值的指标 ,为制定治疗方案和评估预后提供重要参考依据。     

Bone marrow micrometastases in 109 breast cancer patients: Correlations with clinical and pathological features and prognosis

Background: The presence in bone marrow of cells which react with monoclonal antibodies against tumor-associated antigens has been proposed over the last few years as a new prognostic factor in breast cancer patients. Patients and methods: Bone marrow aspirates were obtained from 109 stage I and II breast cancer patients during or 2–4 weeks after primary surgery. The samples were processed for leukocyte separation on a Ficoll-Hypaque gradient and then used to prepare cytospin slides for immunocytochemical analysis. The slides were stained with a pool of monoclonal antibodies (MoAbs) which recognize tumor associated antigens, using the alkaline phosphatase anti-alkaline phosphatase method. The median follow-up was 36 months (range 15–62); 22 patients relapsed and 7 died. Results: Thirty-four of the 109 patients (31.1%) had MoAb positive bone marrow cells. The bone marrow was positive in 28/74 (37.9%) patients who had the aspirate taken during surgery and in 6/35 (17.1%) who had it taken after surgery (p = 0.055). No association was found between bone marrow positivity and tumour size, nodal status, menopausal status, estrogen receptor positivity or the proliferative index. No association was found between bone marrow and prognosis: the log-rank test was 0.291 (p > 0.5) for OS and 0.023 for DFS; the hazard ratio (positive vs negative) was 1.51 for OS (95% CI: 0.33–6.86) and 0.93 for DFS (95% CI: 0.35–2.45). Conclusions: In our series, bone marrow positivity did not correlate with prognostic parameters or prognosis. Of interest is the relative excess of positivity when the bone marrow was obtained during surgery.     

骨髓HuD-mRNA表达在小细胞肺癌微转移检测中的意义

目的 :应用逆转录聚合酶链反应 (RT PCR)法检测确诊小细胞肺癌 (SCLC)患者骨髓、外周血中HuD mRNA的表达 ,分析其临床意义 ,探讨其在检测骨髓微转移中的价值。　方法 :选择病理学确诊为SCLC的初治患者 14例 ,采集骨髓及外周静脉血标本 ,另以 15例非小细胞肺癌 (NSCLC)患者及 12例良性病变患者的骨髓及血标本为对照组。 7例SCLC患者在接受 1～ 2个疗程化疗后再次取骨髓及外周血标本 ,用RT PCR法检测所有标本中HuD mRNA的表达。　结果 :SCLC患者骨髓HuD阳性率 (5 7.1% )显著高于非肿瘤对照组 (16 .7% ) ,P <0 .0 5 ;但与NSCLC对照组差异不明显 (33.3% ) ,P >0 .0 5。SCLC广泛期骨髓HuD阳性率 (85 .7% )高于局限期 (2 8.6 % ) ,P <0 .0 5。外周血标本 :SCLC外周血HuD阳性率 (4 2 .9% )与非肿瘤对照组 (16 .7% )、NSCLC对照组 (33.3% )比较差异均不显著 (均为P >0 .0 5 )。　结论 :SCLC微转移是普遍存在的 ,骨髓微转移的存在与患者的临床分期密切相关。     

Detection of Lymphatic Micrometastases in Patients With Stages I and II Colorectal Cancer: Impact on Five-Year Survival

PURPOSE: Despite having removed the whole macroscopic disease (curative intent surgery), one of five patients with Stages I and II colorectal cancer will develop recurrence. Lymphatic micrometastases detected by immunohistochemistry could be one of explanation for recurrence and cancer-related death in patients without lymph node involvement at light microscopy. However, the biologic importance of micrometastases remains unclear. This study was designed to determine the impact of micrometastases in five-year survival in patients with Stages I and II colorectal cancer.METHODS: This retrospective study included patients operated on between May 1989 and January 1999 for colorectal cancer without histopathologic lymph node involvement. Patients who received any adjuvant therapy were excluded. Immunohistochemical staining of the lymph nodes was performed with antipancytokeratin antibodies. Follow-up data were obtained from the clinical database and death certificates. Survival was estimated by the Kaplan-Meier method and compared by the log-rank test.RESULTS: Micrometastases were observed in 26 of 90 patients (28.9 percent). The mean follow-up time was 90.7 (range, 11–160) months. Seventeen cancer-related deaths occurred during follow-up (18.9 percent), 6 of them in patients with micrometastases (23.1 percent) and 11 in patients without micrometastases (17.2 percent; P = 0.559). Cancer-specific five-year survival was 87 percent in the whole group and 81 percent in patients positive for micrometastases vs. 90 percent in negative patients (P = 0.489).CONCLUSIONS: The presence of micrometastases in patients with Stages I and II colorectal cancer seems not to have any impact on cancer-specific survival.Supported by the Apertus Research Program (Andromaco Pharmaceutical Company) and by The National Public Grant (FONDECYT #1000556).     

Lymph node micrometastases is a poor prognostic factor for patients in pN0 gastric cancer: a meta-analysis of observational studies

Background

There is no consensus as to the impact of lymph node micrometastases (LNMM) on survival of patients with gastric cancer. The aim of this analysis was to investigate the prognostic significance of LNMM in patients with histologic node-negative gastric cancer.

Methods

We searched relevant studies from PubMed, Embase, and the Cochrane Library (1966–2013.5), used software STATA 12.0 to pool the outcomes of each study. Mantel-Haenszel and Inverse Variance methods were used in a fixed effect model and a random effect model, respectively. The hazard ratios (HR) and odds risk (OR) at their 95% confidence intervals (CIs) were used as measures to investigate the prognostic importance of LNMM, by searching for a correlation between the clinical pathologic features and LNMM.

Results

Our analysis of 18 eligible studies revealed that patients with LNMM had an increased likelihood of having a worse 5-y survival rate (HR 2.81; 95% CI：1.96–4.02). Subgroup analyses showed a more significant result for patients in pT1-2N0 (HR 3.52; 95% CI 1.88–6.62). The analyses also revealed that (OR 1.32; 95% CI 1.17–1.48), lymphatic invasion (OR 2.21; 95% CI 1.42–3.44) and venous invasion (OR 1.41; 95% CI 1.08–1.85) were associated with the occurrence of LNMM.

Conclusions

There is a positive correlation between LNMM and an unfavorable surgical outcome in gastric cancer. Undifferentiated histologic findings, lymphatic invasion, and venous invasion are high risk factors for the occurrence of LNMM.     

Detection and Significance of Occult Axillary Metastatic Disease in Breast Cancer Patients

▪ Abstract: After clinical staging, the single most important prognostic factor for patients with newly diagnosed primary breast cancer is the presence or absence of detectable metastases to axillary lymph nodes when examined by conventional light microscopy. More sensitive methods of determination of lymph node status, such as evaluation of serial sections, immunohistochemical staining, and use of molecular biological assays increase the rate of detection of micrometastases. Although the feasibility of enhanced detection of occult axillary metastatic disease is well established, the prognostic significance of such detection is only recently starting to emerge. Furthermore, the enormous recent interest in the application of sentinel lymph node biopsy as an alternative to the evaluation of the entire axilla in patients with breast cancer makes the first-time detailed evaluation for micrometastases practically feasible. In this review the different methods of detecting micrometastatic disease in the axilla and the significance of such findings are discussed. ▪     

A、B期直肠癌淋巴结微转移的研究

目的　研究A、B期直肠癌淋巴结微转移的特点 ,分析微转移与各种临床病理因素及预后的关系。方法　选用鼠抗人细胞角蛋白 2 0 (CK2 0 )单克隆抗体 ,运用免疫组化方法 ,检测 5 7例A、B期直肠癌根治标本的直肠周围淋巴结 183个 ,并进行统计学处理。结果　 2 5例 ( 4 3 .9% )直肠癌的 48个淋巴结 ( 2 6.2 % )出现微转移。淋巴结微转移与原发肿瘤的大小、分化程度、肿瘤的部位有关 ,但与年龄、性别及原发肿瘤侵入的深度无关。随访平均 42个月结束后 ,有微转移的直肠癌患者生存率 ( 74.9% )与无微转移的直肠癌患者 ( 78.8% )无明显差异。结论　直肠癌微转移可能增加局部复发或远处转移的发生 ,但对预后无明显关系