Detection Of Bone Marrow Micrometastases Research Articles

Bone marrow mammaglobin-1 (SCGB2A2) immunohistochemistry expression as a breast cancer specific marker for early detection of bone marrow micrometastases

Despite all the advances in the management of breast cancer (BC), patients with distance metastasis are still considered incurable with poor prognosis. For that reason, early detection of the metastatic lesions is crucial to improve patients’ life span as well as quality of life. Many markers were proposed to be used as biomarkers for metastatic BC lesions, however many of them lack organ specificity. This highlights the need for novel markers that are more specific in detecting disseminated BC lesions. Here, we investigated mammaglobin-1 expression as a potential and specific marker for metastatic BC lesions using our patient cohort consisting of 30 newly diagnosed BC patients. For all patients, bone marrow (BM) aspiration, BM biopsy stained by H&E and BM immunohistochemically stained for mammaglobin-1 were performed. In addition, the CA15-3 in both serum and bone marrow plasma was also evaluated for each patient. Indeed, mammaglobin-1 immuno-staining was able to detect BM micrometastases in 16/30 patients (53.3%) compared to only 5/30 patients (16.7%) in BM biopsy stained by H&E and no cases detected by BM aspirate (0%). In addition, our results showed a trend of association between mammaglobin-1 immunoreactivity and the serum and BM plasma CA15-3. Further validation was done using large publicly available databases. Our results showed that mammaglobin-1 gene expression to be specifically upregulated in BC patients’ samples compared to normal tissue as well as samples from other cancers. Moreover, our findings also showed mammaglobin-1 expression to be a marker of tumour progression presented as lymph nodes involvement and distant metastasis. These results provide an initial evidence for the use of mammaglobin-1 (SCGB2A2) immunostaining in bone marrow as a tool to investigate early BM micrometastases in breast cancer.

Role of Immunohistochemistry in Early Detection of Bone Marrow Micrometastases in Breast Cancer

BackgroundIn Egypt, breast cancer (BC) accounted for 37.7% as reported from WHO in 2008. Mammaglobin (MAM) is BC specific marker, which is almost expressed in breast epithelial cells. It is over expressed in subset of 70–80% of primary and metastatic BC tissues and can be used for identification of BC cells in bone marrow (BM).ObjectivesTo evaluate the significance of immunohistochemical (IHC) method applied on bone marrow trephine biopsy (BMB) in increasing the detection capacity for BC micrometastases in apparently non stage‐IV patients. To compare these results with those obtained by cytological examination of BM aspirate and the different biochemical marrow investigations.Materials and MethodsThirty prospective cases, presented with different stages (0‐III) of BC and who received no chemotherapy, were selected from Alexandria Armed Forces Hospital, Egypt, during the period from March 2014 to December 2015. All patient were subjected to the following work up: Reviewing patients' medical records for all clinical and demographic data. BM cytological examination (at least 6–8 Leishman's‐stained BMA smears) was meticulously scanned for any sheets of non‐haemopoietic malignant cells. Histological examination of BM core biopsy (at least 2.5 cm long); minimum of 2 H&E stained sections were thoroughly examined by two histopathologists for any sheets of nonhaemopoietic malignant cells. Morphological interpretation was guided by the standardized morphological criteria. IHC staining was performed on the BMB sections using MAM monoclonal antibody. CA15‐3 serum and BMA plasma levels were assessed using the fully automated Abbott “Axsym” system. Results 22 patients were diagnosed with infiltrating duct carcinoma (IDC) (73.3%), while the rest (26.7%) were diagnosed with infiltrating lobular carcinoma (ILC). Three patients with IDC were classified as grade I (13.6%), 10 as grade II (45.5%), and 9 as grade III (40.9%). Regarding the stage, 2 patients (6.7%) presented with stage 0, 6 patients (20%) with stage I, 14 patients (46.7%) with stage II and 8 patients (26.7%) with stage III (Figure 1). Examination of the BM aspirates of the 30 patients were completely negative (100%), while examination of the H&E stained sections from the BMB were as follows: 5 patients (16.7%) were positive for BM micrometastases showing hypercellularity, fibrosis, inflammatory infiltration (plasma cells, macrophage and eosinophils), angiogenesis and osteoclastic hyperplasia (Figure 2), while 25 patients (83.3%) were negative. 16/30 cases (53.3%) yielded positive results for the MAM IHC staining on BMB (Figures 3 and 4). There was statistical significant differences between using MAM IHC staining and BMB H&E examination (P=0.0013). By measuring CA15.3 in both serum and BM plasma, there was no statistical significant difference regarding serum and BM plasma CA15.3. Conclusions Superiority of BMB over BMA in detection of BM micrometastatic disseminated tumor cells (DTCs) in patients diagnosed with primary BC. Combining BMB with IHC staining using MAM significantly increases the detection capacity for DTCs in the BM when diagnosing patients with primary BC. In spite of the correlation present between BM plasma and serum levels of CA15‐3, neither CA15‐3 serum levels nor BM plasma levels could be reliable markers for diagnosing or detecting early occult micrometastatic DTCs in the BM at initial diagnosis of primary BC This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Comparative Analysis of Tumor Cell Dissemination to the Sentinel Lymph Nodes and to the Bone Marrow in Patients With Nonmetastasized Colon Cancer

Small nodal tumor infiltrates (SNTI; isolated tumor cells and micrometastases) in sentinel lymph nodes and bone marrow micrometastases (BMM) were independently described as prognostic factors in patients with colon cancer. To examine the association between the occurrence of SNTI and BMM as well as their prognostic relevance. This prospective study was conducted at 3 university-affiliated institutions in Switzerland between May 2000 and December 2006. Statistical analyses were performed in October 2016. A total of 122 patients with stage I to III colon cancer were included. Follow-up time exceeded 6 years, with no patients lost to follow-up. Bone marrow aspiration from the iliac crests and in vivo sentinel lymph node mapping were performed during open standard oncological resection. Bone marrow aspirates were stained with the pancytokeratin marker A45-B/B3. All sentinel lymph nodes underwent multilevel sectioning and were stained with hematoxylin-eosin and the pancytokeratin marker AE1/AE3. Association of SNTI in sentinel lymph nodes and BMM in patients with stage I to III colon cancer and the prognostic effect on disease-free survival (DFS) and overall survival (OS). Of the 122 patients, 63 (51.6%) were female, with a mean (SD) age of 71.2 (11.7) years. Small nodal tumor infiltrates and BMM were found in a total of 21 patients (17.2%) and 46 patients (37.7%), respectively. The occurrence of BMM was not associated with the presence of SNTI by standard correlation (κ, -0.07; 95% CI, -0.29 to 0.14; P = .49) nor by univariate logistic regression analysis (odds ratio, 0.64; 95% CI, 0.22-1.67; P = .37) or multivariate logistic regression analysis (odds ratio, 1.09; 95% CI, 0.34-3.28; P = .88). The presence of SNTI was an independent negative prognostic factor for DFS (hazard ratio [HR], 2.93; 95% CI, 1.24-6.93; P = .02) and OS (HR, 4.04; 95% CI, 1.56-10.45; P = .005), as was BMM (HR, 2.07; 95% CI, 1.06-4.06; P = .04; and HR, 2.68; 95% CI, 1.26-5.70; P = .01; respectively). The combined detection of BMM and SNTI demonstrated the poorest DFS (HR, 6.73; 95% CI, 2.29-19.76; P = .006) and OS (HR, 5.96; 95% CI, 1.66-21.49; P = .03). This study demonstrates no association between the occurrence of SNTI and BMM in patients with stage I to III colon cancer. However, both SNTI and BMM are independent negative prognostic factors regarding DFS and OS, and the occurrence of both is associated with significantly worse prognosis compared with either one of them. clinicaltrials.gov Identifier: NCT00826579.

Prognostic Value of Bone Marrow Micrometastasis in Patients with Operable Esophageal Squamous Cell Carcinoma: A Long-Term Follow-Up Study

Detection of bone marrow micrometastasis (BMM) has been focused on as a prognostic parameter in various malignant neoplasms recently. This study was designed to evaluate the prognostic significance of BMM detection in patients with operable esophageal squamous cell carcinoma (ESCC) after long-term follow-up. In 61 consecutive patients with ESCC who had undergone radical surgical resection, BMM was detected through reverse transcriptase-polymerase chain reaction (RT-PCR). Correlation between BMM detection and prognosis of the 61 patients was analyzed. BMM was found in 13 patients (21.3%). No significant correlation between BMM detection and tumor, node, metastasis (TNM) stage was found. The median survival time, 5-year overall survival rate, 5-year disease-free survival rate, and 5-year distant disease-free survival rate for cases with positive BMM were 13.0 months, 15.4%, 7.7%, and 34.2%, respectively, compared with that of 66.0 months, 59.7%, 49.1%, and 60.6% for cases with negative BMM (p < 0.05). In multivariate analysis, BMM were found to be an independent factor in the prediction of overall survival (odds ratio [OR] 3.928, p = 0.001), disease-free survival (OR 4.285, p < 0.001), and distant disease-free survival (OR 3.270, p = 0.013). BMM is an independent prognostic factor in the prediction of the subsequent development of metastatic disease and disease outcome for operable ESCC patients, and may be a useful adjunct to conventional tumor staging. Further studies are required to evaluate the value of neoadjuvant or adjuvant systemic therapy in ESCC patients with BMM.

Bone marrow micrometastases in esophageal carcinoma: a 10-year follow-up study

Detection of bone marrow micrometastases (BMMs) in patients with esophageal carcinoma may indicate a metastatic phenotype. We assessed if the presence of BMMs had adverse prognostic significance in a 10-year follow-up study. Patients undergoing surgery for esophageal cancer were prospectively recruited between February 1999 and August 2000. Bone marrow aspirates were obtained from the iliac crest of patients under general anesthesia at the time of surgery. Immunocytochemical analysis using anticytokeratin antibodies CAM 5.2 and AE1/AE3 was undertaken to determine the presence of BMMs. Union International Contre le Cancer staging was recorded for all patients. Patient follow-up was completed over a 10-year period through analysis of the Northern Ireland Cancer Registry. Forty-two patients (male = 35) were included, with a mean age of 67.2 years (range 39-83). BMMs were detected in 19 patients (45.2%). International Contre le Cancer tumor staging was stage I = 6, stage II = 10, stage III = 24, and stage IV = 2. BMMs were associated with lymphovascular invasion (P= 0.02) and advanced T stage (P= 0.02). Overall, 10-year survival was 21.4% (n= 9), with a median follow-up of 877.5 days (interquartile range 391.5-2546.3). There was no statistically significant difference between the survival of patients with or without BMMs (1451.4 vs. 1431.6 days, P= 0.99). Univariate analysis demonstrated a trend toward decreased survival for patients with positive lymph nodes (P= 0.07), an increased T stage (P= 0.06), and lymphovascular invasion (P= 0.07). Multivariate analysis demonstrated that none of the variables were significant predictors of mortality. Although the presence of BMMs correlates with recognized adverse tumor characteristics in patients with esophageal cancer, micrometastases detected in the bone marrow at time of surgery does not influence long-term survival.

Flow cytometric detection of micrometastases to rib marrow in patients with non-small lung cancer

Objective To evaluate the detection value of rib bone marrow micrometastasis ( BMM ) in patients with non-small lung cancer ( NSCLC ) by using flow cytometry ( FCM ) and determine the relationship between BMM and clinicopathological parameters. Methods 158 patients undergoing surgical treatment for NSCLC during April 2004 to May 2007 were prospectively analyzed. Bone marrow aspirates were obtained intraoperatively and the expression rate of the postive cells was determined by using flow cytometry. Results 59of the 158 ( 37.34 ％ ) patients occurred, 18 ( 30.51％ ) of whom developed distant metastases. 14 of the 99 ( 13.9％ ) patients without BMM developed distant metastases.BMM was correlated with tumor size and tumor stage, but not with age, sex, pathological types, differentiation degrees. The bigger the tumor size and the higher the tumor stage, the higher the frequency of BMM.Patients with BMM had a higher metastatic rate than in those without BMM ( P ＜ 0.05 ). Conclusions Non-small lung cancer can occur early bone marrow micrometastasis. BMM is correlated with some clinic pathological parameters. Detection of BMM by flow cytometry can be used as an indicator for instruction of treatment and prediction of prognosis. Key words: Flow cytometry; Non-small lung cancer; Bone marrow micrometastasis

Bone marrow micrometastases in advanced stage non-small cell lung carcinoma patients

The clinical relevance of bone marrow micrometastases in non-small cell lung cancer (NSCLC) is undetermined, and the value of such analyses in advanced stage patients has not been assessed previously. Immunomagnetic selection with the MOC31 (anti-EpCam) antibody was performed to isolate and detect tumor cells in bone marrow aspirates obtained from 196 patients with NSCLC, and the patients were subjected to follow-up for the assessment of survival. Repeated bone marrow samples, 2-7 samples per patient, were obtained from 13 long-term survivors. MOC31 positive tumor cells were detected in 107 of 196 (55%) samples, a frequency similar to results reported in low-stage patients prior to curative surgical resection for NSCLC. No association was found between the presence of bone marrow micrometastases and disease stage or histological subgroup, and survival was similar in patients with and without detectable tumor cells in bone marrow. Repeated bone marrow analysis revealed, for the first time in this group of patients, the continued presence of tumor cells regardless of the given therapy and treatment response. The immunomagnetic method utilized is a feasible strategy for the detection of bone marrow micrometastases in NSCLC. In advanced stage patients, the presence of MOC31 positive cells in bone marrow does not predict survival. Repeated analyses of bone marrow samples from long-term survivors revealed tumor cells in bone marrow which may represent dormant cells of particular interest for further characterization and follow-up.

American Society of Clinical Oncology 2007 Update of Recommendations for the Use of Tumor Markers in Breast Cancer

To update the recommendations for the use of tumor marker tests in the prevention, screening, treatment, and surveillance of breast cancer. For the 2007 update, an Update Committee composed of members from the full Panel was formed to complete the review and analysis of data published since 1999. Computerized literature searches of MEDLINE and the Cochrane Collaboration Library were performed. The Update Committee's literature review focused attention on available systematic reviews and meta-analyses of published tumor marker studies. In general, significant health outcomes (overall survival, disease-free survival, quality of life, lesser toxicity, and cost-effectiveness) were used for making recommendations. Recommendations and Thirteen categories of breast tumor markers were considered, six of which were new for the guideline. The following categories showed evidence of clinical utility and were recommended for use in practice: CA 15-3, CA 27.29, carcinoembryonic antigen, estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2, urokinase plasminogen activator, plasminogen activator inhibitor 1, and certain multiparameter gene expression assays. Not all applications for these markers were supported, however. The following categories demonstrated insufficient evidence to support routine use in clinical practice: DNA/ploidy by flow cytometry, p53, cathepsin D, cyclin E, proteomics, certain multiparameter assays, detection of bone marrow micrometastases, and circulating tumor cells.

Correlation of bone marrow micrometastases with nodal status in gastrointestinal tumors

14577 Background: Presence of bone marrow micrometastases (BMM) is an important prognostic factor for patients (pts) with solid tumors. Sentinel lymph node (SLN) mapping (M) has been found to upstage pts with gastrointestinal (GI) malignancies. However, a direct correlation between the presence BMM and nodal metastases (mets) is lacking. Hence, a retrospective study was undertaken to determine the relationship between BMM and nodal status in various GI malignancies. Methods: A total of 159 consecutive pts with GI malignancies were analyzed. Of these, 6 pts with non-epithelial tumors were excluded. All pts underwent bilateral posterior-superior-iliac spine bone marrow (BM) aspiration. BM samples were examined for micrometastases by Automated Cellular Imaging System using ChromaVision Cytokeratin Detection Kit to detect cells expressing CK8 as defined by CAM 5.2 monoclonal antibody. When feasible, SLNM was performed followed by standard oncologic resection (n=105). The SLN were examined by H&amp;E and immunohistochemical (IHC) staining. Results: A total of 153 pts were analyzed in the study. Of these 13.72% (21/153) were found to have BMM (CI 8.70–20.21, p=0.05). When analyzed by sites, the incidence of BMM was 33.3% (4/12) for stomach cancer, 9.1% (1/11) for pancreatic cancer, 12.36% (11/89) for colon cancer and 16.13% (5/31) for rectal cancer. No BMM were detected in pts with cancer of esophagus (n=7), small bowel (n=2) and liver (n=1). The BMM was positive bilaterally in 61.9% (13/21), and unilaterally in 38.1% (8/21). In pts who underwent SLNM (n=105), BMM were detected in 11.11% of SLN positive pts vs. 10.14% for SLN negative pts (p=0.87). For a subgroup of pts with colorectal cancer who underwent SLNM (n=97), BMM were detected in 9.67% of SLN positive pts vs. 9.09% for SLN negative pts (p=0.92) Conclusions: BMM did not correlate with nodal status for GI malignancies suggesting a possible different mechanism for metastases. Detection of BMM may have a significant clinical value in SLN negative pts who may benefit from adjuvant therapy. [Table: see text] No significant financial relationships to disclose.

Recent advances in the detection of bone marrow micrometastases: A promising area for research or just another false hope? A review of the literature

The presence of early disseminated tumor cells (DTC), otherwise termed micrometastases or minimal residual disease, in the bone marrow (BM), or other secondary compartments, such as the blood and the lymph nodes, is the main reason for recurrence of patients with early stage epithelial cancers after "curative" resection of the primary tumor. There is increasing evidence, that the detection of DTC in BM aspirates may provide additional and independent prognostic information and aid in the stratification of these patients for adjuvant clinical treatment. However, the clinical relevance of micrometastases has not yet been firmly established. In addition, the molecular events and interactions that prevail in early metastatic disease and determine the formation or not of overt metastases remain poorly understood. The methods currently used for the detection of micrometastatic cells include extremely sensitive immunocytochemical and molecular assays, often in conjunction with enrichment techniques for the purification of tumor cells and additional increase of their sensitivity. Nevertheless, the specificity of these methods is mostly inadequate. After the impressive advances of molecular cytogenetics, a highly accurate and global assessment of the genetic status of tumors is now possible. Therefore, the greatest challenge of our time is the application of these novel technologies for the clarification of the key molecular events that initiate metastatic spread. This will further enable us to identify the highly specific and sensitive diagnostic and prognostic markers as well as the therapeutic targets which are so urgently needed.

Detection of bone marrow micrometastasis and microcirculating disease in rhabdomyosarcoma by a real-time RT-PCR assay

To assess if molecular detection of minimal disseminated disease by real-time reverse transcription and polymerase chain reaction (RT-PCR) could contribute to a better treatment stratification in patients with rhabdomyosarcoma (RMS). Relative quantification of the tumor-mRNA present in serial samples of bone marrow (BM) and peripheral blood (PB) from 16 patients with RMS (7 alveolar and 9 embryonal) was performed by a real-time RT-PCR assay. Expression of MyoD1 and acetylcholine receptor (AChR) was analyzed in all samples, along with PAX3/7-FKHR in samples from alveolar tumors. A good correlation was found between the expression of PAX3/7-FKHR and AChR, while MyoD1 was more sensitive but less specific. In this study, patients with positive PB at the end of treatment showed a poorer prognosis than patients with negative PB. Moreover, in this patient cohort, metastatic relapses were preceded by the detection of microcirculating disease in all cases. The detection of minimal circulating and micrometastatic disease by real-time RT-PCR, based on the expression of multiple genes, yields highly reproducible results. Patients with positive PB after treatment show poorer survival than patients without microcirculating disease.

The value of bone marrow aspirates culture for the detection of bone marrow micrometastasis in breast cancer

Background. - Detection of micrometastasis is an important problem of clinical significance for a better understanding and control of tumor progression, which will improve patients' survival time. Tumor cells in bone marrow (BM) aspirates are indicative of the general disseminative metastasis in patients with early breast cancer and characterization of breast cancers by various tumor markers which are appropriate for the identification of high risk groups. Materials and methods. - Bone marrow aspirates were obtained from 44 breast cancer patients at the time of surgery. To identify micrometastases in bone marrow, an immunocytochemical assay for epithelial cytokeratin (CK) was performed at the second passage after selective culture. Cytokeratin-positive bone marrow disseminated cancer cells were observed in more than 90% of the patients. This high incidence needs further investigation with bigger sample size to confirm. However, these results indicate that this technique can be used as an early diagnostic technique of bone marrow micrometastases in the patient with breast cancer thereby promoting the development of therapeutic strategy. High incidences need further investigation with bigger samples to confirm.

Detección de micrometástasis en médula ósea de pacientes con cáncer de vesícula biliar

There is a very strong documented correlation between the appearance of cancer cells in blood and occurrence of metastasis in gastrointestinal cancer. To determine MUC1, CK19, CK20 and CEA mRNA expression in bone marrow of patients with gallbladder cancer and evaluate its clinical significance. Sixty eight samples were analyzed, 38 bone marrow samples of gallbladder cancer patients, 20 healthy donors, and 10 frozen samples of gallbladder cancer. Nested reverse transcriptase-polymerase chain reaction (nested RT-PCR) was used to analyze mRNA expression. All frozen tumors were positive for CEA, CK19, and MUC1 mRNA and 70% were positive for CK20. Seventeen of 20 donor samples were positive for MUC1 and only one sample from donors was positive for both CK20 and CK19 mRNA. Among the 38 blood and bone marrow samples of gallbladder cancer patients, the expression of MUC1, CK19, CK20, and CEA, mRNA was 60.5% (23/38), 31.6% (12/38), 7.9% (3/38), and 7.9% (3/38), respectively. Disregarding the MUC1 results. 37% (14/38), 13% (5/38) and 5% (2/38) were positive for one, two and three markers respectively. Not significant differences were found in survival with a follow up to 12 months. Our results indicate that the molecular detection of tumor cells in bone marrow in patients with gallbladder carcinoma is technically possible, being CEA, CK19 and CK20 gene expression the best markers. The MUC1 gene expression marker was highly unspecific and it should not been considered. The detection of bone marrow micrometastasis might be helpful in prognosis and the selection of clinical treatment but a larger series with a longer follow-up should be studied.

Bone Marrow Micrometastases and Markers of Angiogenesis in Esophageal Cancer

Background Tumor angiogenesis is critical for metastasis development. The detection of bone marrow micrometastases may indicate a metastatic phenotype. We aim to establish if the detection of bone marrow micrometastases associates with elevated markers of angiogenesis and adverse histopathologic features of esophageal cancer. Methods Bone marrow aspirates from 49 patients with esophageal cancer were assessed and assigned to be positive or negative for micrometastases. Routine histologic assessment of the primary tumor was also undertaken. Circulating and tumor levels of the angiogenic cytokine vascular endothelial growth factor were determined in plasma and tumor homogenate. Intratumor microvessel density was evaluated by counting anti-CD34 positive neovessels. Results Twenty-two patients were positive for bone marrow micrometastases (44.9%). The detection of micrometastases was associated with advanced T stage (T3/4 vs T1/2; p = 0.023), circumferential margin involvement ( p = 0.002) and lymphovascular invasion ( p = 0.024). Plasma vascular endothelial growth factor was significantly more elevated in micrometastatic-positive patients than in those without micrometastases ( p = 0.018). No difference was noted in tumor vascular endothelial growth factor expression. For adenocarcinomas alone, intratumor microvessel density was significantly higher in micrometastatic positive cases ( p = 0.03). This was not the case for squamous cell carcinomas. Conclusions The detection of bone marrow micrometastases is associated with esophageal tumors of advanced T stage and specifically for adenocarcinomas with tumor vascularity. Plasma vascular endothelial growth factor is elevated in micrometastatic positive cases and might be derived from sources other than the primary tumor.

Sentinel Lymph Node Biopsy or Detection of Micrometastasis in Bone Marrow: Which Might Be an Alternative to Axillary Lymph Node Dissection in Breast Cancer Patients?

Axillary lymph node status has limited prognostic significance in breast cancer patients and much improvement can be made. Sentinel lymph node biopsy is emerging as an alternative to axillary lymph node dissection for staging, but its prognostic relevance is still uncertain. Detection of micrometastases in sentinel nodes and bone marrow may provide more information, but the clinical significance still needs to be confirmed by ongoing large trials. In this review, we focus on the possibility of sentinel lymph node biopsy or detection of bone marrow micrometastasis replacing traditional axillary lymph node dissection.

Detection of bone marrow micrometastasis in esophageal cancer patients by immunomagnetic separation

Detection of micrometastasis to the bone marrow can predict widespread disease and a poor prognosis of cancer patients. The purpose of this study was to evaluate the clinical significance of detecting micrometastasis in the bone marrow of esophageal cancer patients. Bone marrow and peripheral blood samples were obtained from 52 squamous esophageal cancer patients at the time of surgery. These samples were enriched by immunomagnetic separation and immunostained with an anti-cytokeratin antibody. Expression of vascular endothelial growth factor (VEGF) and cyclin D1 was examined in the primary tumors. Cytokeratin-positive cancer cells were observed in the bone marrow of 13 (25%) out of 52 patients. Among them, three patients also had cancer cells in the peripheral blood. The presence of bone marrow micrometastasis was correlated with lymph node metastasis (pN) but not associated with depth of tumors (pT). Detection of bone marrow micrometastasis was significantly correlated with VEGF expression of the primary tumors. Cumulative survival of patients with bone marrow micrometastasis was significantly lower than that of patients without it (p=0.0008). Hematogenous recurrence was more frequent in patients with bone marrow micrometastasis than in those without it (p=0.0004). Three patients who had local or dissemination recurrence did not have bone marrow micrometastasis. Detection of cancer cells in the bone marrow might be an indicator of early hematogenous metastasis in esophageal cancer patients. Intensive postoperative chemotherapy seems to be indicated for these patients.

Detection of bone marrow micrometastasis in gastric cancer patients by immunomagnetic separation.

Micrometastasis to the bone marrow can predict widespread disease and a poor prognosis of cancer patients after surgery. The purpose of this study was to evaluate the clinical significance of detecting micrometastasis in the bone marrow of gastric cancer patients. Bone marrow and peripheral blood samples were obtained from 53 gastric cancer patients at the time of surgery. These samples were enriched by immunomagnetic separation and immunostained with an anti-cytokeratin antibody. Expression of vascular endothelial growth factor and erbB-2/HER2 was examined in the primary tumors. Cytokeratin-positive cancer cells were observed in the bone marrow of 16 (30%) of 53 patients. Among them, two patients also had cancer cells in the peripheral blood. The presence of bone marrow micrometastasis was correlated with the depth of invasion and lymph node metastasis but was not associated with peritoneal dissemination. Detection of bone marrow micrometastasis was not correlated with vascular endothelial growth factor or HER2 expression in the primary tumors. Four patients with micrometastasis had recurrence in the liver or lungs, but this did not occur in patients without micrometastasis. Detection of cancer cells in the bone marrow might be an indicator of postoperative hematogenous metastasis in gastric cancer patients.

Isolation of Circulating Cancer Cells From Whole Blood By Immunomagnetic Cell Enrichment and Unenriched Immunocytochemistry In Vitro

Isolation of Circulating Cancer Cells From Whole Blood By Immunomagnetic Cell Enrichment and Unenriched Immunocytochemistry In Vitro

Clinical study on the bone marrow micrometastases in patients with non-small cell lung cancer by immunohistochemical techniques

To investigate the immunohistochemical detected method and the clinical incidence of the bone marrow micrometastases (BMM) in patients with non-small cell lung cancer (NSCLC) and to analyze the sensitivity and specificity and clinical application value. Bone marrow samples were collected from the anterior superior iliac spines or posterior superior iliac spines of 53 patients with NSCLC in clinical stage I to III and 15 patients in stage IV, and the BMM was detected by immunohistochemical techniques (IHC) using monoclonal antibodies AE1/AE3 against cytokeratin. Chi-square test was used statistically. The IHC sensitivity could be 10⁻⁵. The BMM positive rate was 22.6% (12/53) in stage I to III and 53.3% (8/15) in stage IV, and there was a significant difference in the BMM positive rate between stage I to III and stage IV (P < 0.05). No correlation was observed between BMM and sex, age, KPS, pathology classification and cancer cell differentiation. The detection of BMM by IHC is convenient, sensitive, and specific. It might be helpful to diagnose bone marrow micrometastasis in patients with NSCLC.

Comparison of detection of micrometostases in rib marrow aspirates compared to iliac crest aspirates

Iliac crest marrow remains the standard sampling site for detection of bone marrow micrometastases in breast cancer patients.