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PubMed2006

Soluble VCAM-1 and E-selectin in breast cancer: relationship with staging and with the detection of circulating cancer cells.

In breast cancer, the correct evaluation of cancer dissemination is essential to establish prognosis and treatment choices. This study analyses the relationship between circulating levels of soluble VCAM-1 and E-selectin and the presence of circulating cancer cells in breast cancer patients. Plasma levels of VCAM-1 and E-selectin were measured by enzyme-linked immunosorbent assay (ELISA). The presence of circulating cancer cells was diagnosed using a RT nested-PCR assay detecting the cancer specific transcript, epidermal growth factor receptor variant III (EGFRvIII) mRNA. Blood samples were collected from 64 patients divided in three groups: group A of 11 women selected for neoadjuvant chemotherapy; group B of 13 women with metastatic disease and group C, with 40 women having completed their treatment at least one year ago and with no evidence of relapse. The mutant transcript was detected in 45.5% of patients from group A, in 61.5% of patients from group B and in none of the group C patients. For both VCAM-1 and E-selectin, plasma levels increased with disease staging and with the presence of EGFRvIII mRNAin peripheral blood. The differences were statistically significant (p < 0.025) when group C was compared with all patients from group B, with patients from group B with EGFRvIII positive results or with all patients with EGFRvIII positive results. Increased plasma levels of VCAM-1 and E-selectin are associated with advanced stage of breast cancer and with the presence of circulating cancer cells. The combined analysis of these parameters may contribute to a more accurate evaluation of cancer dissemination.

Antineoplastic Agents↗

PubMed1980

A microstereotactic approach to small CNS lesions. Part II; Adjuvant therapy. Clinical potential and toxicity of intraventricular lymph infusion.

A technique is described for adjuvant therapy of small central nervous system tumors after their removal. The feasibility of direct application of lymphocytes from the thoracic duct to the ventricular system of cats has been explored. Reduction of the antigenic mass by techniques described in part I of this paper is probably crucial. Patients with gliomas are known to have circulating tumor-specific lymphocytes which may not have direct access to the CNS. This study demonstrated no toxicity from the chronic intraventricular infusion of thoracic duct lymph in cats.

Animals↗

PubMed1962

A study in isolating circulating cancer cells from blood.

Samples of blood from 49 cancer cases were examined for free cancer cells. Cells identified as cancer cells were found in the blood of four. Cells which looked highly suspicious of being cancer cells were isolated from the blood of nine other cases. No conclusion was possible regarding the type of cells in eight cases. This illustrates the difficulty of cytological diagnosis and shows the need for new methods of identification.

Cell Biology↗

PubMed1995

Association between tumour angiogenesis and tumour cell shedding into effluent venous blood during breast cancer surgery.

Tumour angiogenesis is a powerful prognostic indicator in breast cancer. Shedding of tumour cells into the bloodstream is essential for haematogenous metastasis. The relation between cell shedding and angiogenesis in human cancer is not known. We studied vascular density and cell shedding in 16 women undergoing breast cancer surgery. Circulating cells were found in one patient before, in six during, and none after operation. Vascular density was related to detection of circulating tumour cells during surgery (correlation coefficient for first intraoperative sampling = 0.56, p = 0.024, 95% CI 0.08 to 0.83). Tumour cell shedding during surgery is related to vascular density; the relevance of this relation to the prognostic significance of vascular density in breast cancer merits further study.

Breast Neoplasms↗

PubMed1992

Circulating prostate specific antigen-positive cells correlate with metastatic prostate cancer.

Analytical flow cytometry was used to study circulating prostate specific antigen (PSA)-positive cells in 40 consecutive patients with newly diagnosed, untreated prostate cancer; 25 patients (63%) had metastatic disease confirmed by a positive bone scan. Cell suspensions were prepared for each patient from both the primary tumour and peripheral blood samples. The cells were stained with a monoclonal antibody against PSA, and analysed by flow cytometry; PSA-positive cells were sorted according to their immunofluorescence and light scatter properties. The cellular deoxyribonucleic acid (DNA) content of each specimen was also analysed to establish ploidy status. PSA-positive cells were detected in the peripheral blood of 33 patients (83%). The presence of these cells in the circulation showed a higher degree of sensitivity and specificity in predicting positive bone scans than did serum PSA levels. Circulating PSA-positive cells may represent either a subpopulation of tumour cells with distinct metastatic properties or, alternatively, host immunocytes which take up PSA in an active or passive manner.

Aged↗

PubMed2006

Prognostic value of RT-PCR tyrosinase detection in peripheral blood of melanoma patients.

Malignant melanoma (MM) prognosis has been related to tumour thickness and clinical stage and metastasis risk has been associated with presence of tumour cells in peripheral blood. The aim of this study was to determine the relationship between presence of tyrosinase in peripheral blood of MM patients and their clinical prognosis. Blood samples from 58 MM patients (stage I-IV) were analysed, using RT-PCR assay to detect tyrosinase mRNA. The results showed that positive RT-PCR assay for tyrosinase were significantly associated with clinical status and tumour thickness. After a median follow-up of 24 months, RT-PCR results were found to be significant correlated with recurrence (p<0.05) and clinical stage III (p<0.05). Separate analysis of stage III tumours to determine the prognostic value of tyrosinase presence in peripheral blood showed an overall 24-month survival rate of 70% in the RT-PCR negative group versus 10% in the positive group (p<0.02). These results suggest that detection of circulating melanoma cells may be especially relevant in stage III patients, in whom RT-PCR positivity defines a subpopulation at high risk of recurrence.

Adult↗

PubMed1997

Detection of circulating melanoma cells by specific amplification of tyrosinase complementary DNA is not a reliable tumor marker in melanoma patients: a clinical two-center study.

PURPOSE: Recently, the reverse-transcription polymerase chain reaction (RT-PCR) of tyrosinase messenger RNA (mRNA) was reported to be a useful tool for the detection of circulating tumor cells in the peripheral blood of melanoma patients. Our aim was to evaluate critically the diagnostic value of this marker by investigating a significant number of patients in different stages of the disease in a two-center study. PATIENTS AND METHODS: Different techniques of blood collection, RNA isolation, and RT-PCR were compared, and the detectability of tyrosinase mRNA was tested using nine different melanoma cell lines. The sensitivity of the method was confirmed by blood spiking experiments and the specificity by restriction enzyme analysis. Subsequently, a total of 153 blood samples from 137 individuals (30 healthy subjects, five basal cell carcinoma, and 102 melanoma patients) were investigated. RESULTS: The detection level of melanoma cells differed between the cell lines tested. However, we could reproducibly detect single melanoma cells by spiking whole blood samples from healthy volunteers. One of 43 patients with primary melanoma (2.3%), zero of 15 patients with regional metastasis (0%), and 12 of 44 patients with advanced disease (27.3%) were found to be RT-PCR positive. All blood samples obtained from controls and patients with basal cell carcinoma were tyrosinose mRNA negative. CONCLUSION: Our data support the recent doubts that the detection of circulating tumor cells in melanoma patients using the tyrosinase mRNA RT-PCR is not sensitive enough to be used either as a melanoma progression marker in early stages of the disease or to monitor therapy in advanced stages of the disease.

Biomarkers, Tumor↗

PubMed1998

Detection of germ cell tumor cells in apheresis products using polymerase chain reaction.

The contamination of apheresis products with tumor cells was evaluated in patients undergoing autologous peripheral blood stem cell transplantation for germ cell tumors. A blinded, retrospective analysis was performed on 63 apheresis products from 28 patients using the PCR and primers for beta human chorionic gonadotropin (beta-HCG). Of the 20 patients with beta-HCG-secreting tumors, 8 apheresis products from 7 patients were PCR positive. PCR was negative in the 8 patients whose tumors did not secrete beta-HCG. Twenty-two apheresis products from patients with lymphoma and breast cancer were negative for beta-HCG expression. Evaluating the 20 patients with beta-HCG-secreting tumors, 100% of PCR-positive patients had elevated serum beta-HCG at the time of apheresis compared to 46.2% of PCR-negative patients (P = 0.04). A positive PCR was also associated with a higher serum beta-HCG at diagnosis (P = 0.03). Patients receiving a PCR-positive product had a higher relapse rate (85.7 versus 61.5%) and were more likely to have visceral metastasis (100 versus 61.5%), although the numbers did not reach statistical significance (P = 0.35 and 0.11, respectively). The finding of beta-HCG mRNA in apheresis products strongly suggests the presence of circulating tumor cells in a significant number of germ cell patients undergoing autologous transplantation. This assay may be useful in monitoring attempts at tumor cell depletion and in developing improved prognostic models for assessing risk of relapse after transplantation.

Blood Component Removal↗

PubMed2005

Lymphangiogenesis-mediated shedding of LAM cell clusters as a mechanism for dissemination in lymphangioleiomyomatosis.

Lymphangioleiomyomatosis (LAM) affects exclusively women of reproductive age, involves the lungs and axial lymphatic system, and is frequently complicated with renal angiomyolipomas. LAM lesions are generated by the proliferation of LAM cells with mutations of one of the tuberous sclerosis complex (TSC) genes. Recent studies indicate that LAM cells can migrate or metastasize to form new lesions in multiple organs, although they show a morphologically benign appearance. In the previous study, we reported LAM-associated lymphangiogenesis and implicated its role in the progression of LAM. In this study, we further focused on the lymphatic abnormalities in LAM: LAM-associated chylous fluid (5 pleural effusion and 2 ascites), surgically resected diaphragm (1 patient), and axial lymphatic system including the thoracic duct, lymph nodes at various regions, and diaphragmatic lymphatic system (5 autopsy cases). We demonstrated that LAM cell clusters enveloped by lymphatic endothelial cells (LCC) in all chylous fluid examined. We identified LAM lesion in the diaphragm (2 of 5 autopy cases and one surgical specimen), thoracic duct (5 of 5), and lymph nodes (retroperitoneal (5 of 5), mediastinal (4 of 5), left venous angle (5 of 5) with total positive rate of 68% to 88% at each region of the lymph node, but less frequent or none at remote lymph nodes located away from the axial lymph trunk (cervical [1 of 5] and axillary [0 of 5]). LCCs were identified in intra-LAM lesional lymphatic channels where LAM cells proliferate along lymphatic system. In in vitro culture system, LCC can fragment into each proliferating LAM cell. These findings suggest that LAM-associated lymphangiogenesis demarcates LAM lesion into bundle- or fascicle-like structure and eventually shed LCC into the lymphatic circulation and that LCCs play a central role in the dissemination of LAM lesion.

Adult↗

PubMed2006

Perioperative cyclooxygenase 2 inhibition to reduce tumor cell adhesion and metastatic potential of circulating tumor cells in non-small cell lung cancer.

OBJECTIVE: Surgical manipulation of lung cancers may increase circulating tumor cells and contribute to metastatic recurrence after resection. Cyclooxygenase 2 is overexpressed in most non-small cell lung cancer and upregulates the cell adhesion receptor CD44. Our goal was to examine the effects of perioperative cyclooxygenase blockade on the metastatic potential of circulating tumor cells, CD44 expression, and adhesion of cancer cells to extracellular matrix. METHODS: Human non-small cell lung cancer cells (A549) were injected through the lateral tail vein in an in vivo murine model of tumor metastasis with three random treatment groups: no treatment, perioperative selective cyclooxygenase 2 inhibition (celecoxib) only, and continuous celecoxib. Lung metastases were assessed at 6 weeks by a blinded observer. For in vitro experiments, cells were treated with celecoxib, and expression of CD44 was determined by Western blotting. Extracellular matrix adhesion was assessed by Matrigel (BD Labware, Bedford, Mass) assay. RESULTS: In vivo lung metastases were significantly decreased relative to control by both perioperative and continuous celecoxib (P = .0135). There was no significant difference in number of metastases between continuous and perioperative treatment groups. In vitro adhesion to the extracellular matrix was significantly inhibited by celecoxib in a dose-dependent manner (P < .01). A549 cells expressed high levels of CD44, upregulated by interleukin 1beta and downregulated by celecoxib. CONCLUSION: Celecoxib significantly reduced establishment of metastases by circulating tumor cells in a murine model. It also inhibited CD44 expression and extracellular matrix adhesion in vitro. Perioperative modulation of cyclooxygenase 2 may be a novel strategy to minimize metastases from circulating tumor cells during this high-risk period.

Adenocarcinoma↗

PubMed1983

[Role of blood platelets and prostaglandins in blood-borne metastases].

Coagulation-fibrinolysis system, especially blood platelets, plays an important role in the stage of lodgement of circulating tumor cells. In this paper, we discuss that some tumor cells induce platelet aggregation in vitro and several anti-platelet drugs inhibit not only platelet aggregation in vitro but also tumor blood-borne metastases in vivo. For the therapy against cancer metastases, the role of blood platelets in blood-borne metastases should be clarified.

Animals↗

PubMed1998

[Biologic detection methods in the comparison of circulating tumor cells and micrometastases].

Early studies could not prove any diagnostic or prognostic value of the presence of tumor cells in the circulation. Recent knowledge in the field of molecular and cellular pathology provided better understanding of mechanisms of metastasis formation therefore advanced detection of circulating cancer cells has been suggested as a supplementary method of staging metastatic cancer. Beside the widely used immunocytochemical methods the reverse transcriptase-polymerase chain reaction (RT-PCR) is now the most relevant technique in studying micrometastases of solid tumors. Magnetic activated cell sorting (MACS) is a recently developed method for the enrichment of different cells from suspensions by magnetic labelling of their surface antigens. RT-PCR seems to be the most sensitive to detect circulating cancer cells or micrometastases, but it is possible by MACS to purify cells for further immunological, biochemical or genetic analysis. The aim of this review is to give a brief summary of recently used methods and to discuss the clinical relevance of the attainable results.

Biomarkers, Tumor↗

PubMedDate not supplied

Evaluation of circulating tumour cells expressing CD44 variants in the blood of gastric cancer patients by flow cytometry.

The occurrence of circulating tumour cells in the blood of 51 patients with gastric cancer (stages I-IV) was studied using flow cytometry, cell sorting, immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). The lysed whole blood samples were stained with monoclonal antibodies against common leukocyte antigen (CD45), epithelial membrane antigen (EMA), tumour associated glycoprotein 72kD (TAG72), CD44 variants (v5 and v6) and analysed by flow cytometry within ungated or CD45-gated populations. The frequency of detection of TAG72+, CD44v5+ and v6+ cells within CD45- gate was considerably increased in comparison to the ungated population. Furthermore, the presence of tumour cells was directly demonstrated by immunostaining for cytokeratin 18 of sorted CD45- population. The presence of CD44v5+, v6- cells and CD44v-mRNA in the blood was compared to their expression in the primary tumour. The occurrence of circulating CD44v+ cells was associated with their presence in the primary tumour and CD44v-mRNA in the blood. The method described may provide a sensitive tumour marker-independent tool for detection of circulating tumour cells in cancer patients.

Adult↗

PubMed2002

The relationship between gastric cancer cells circulating in the blood and microsatellite instability positive gastric carcinomas.

BACKGROUND: Cancers characterized by microsatellite instability may be biologically different from their counterparts with stable microsatellite sequences. Circulating cancers cell present in blood prior to surgery may constitute an adverse prognostic finding. AIM: To correlate these two phenomena with morphological features and survival in advanced gastric cancer. METHODS: We examined 76 cases of resected sporadic, advanced gastric cancer by means of routine morphology and a panel of microsatellite markers. Sixty-six cases were screened for presence of cancer cells circulating in blood prior to the surgery using combined morphological and immunocytochemical approach. RESULTS: Twenty-one (27.6%) cases demonstrated microsatellite instability in at least one locus. Among them 11 (14.5%) showed microsatellite instability in more than 30% (4/12) examined loci, and they were therefore designated as replication error positive (RER+). Circulating cancer cells were detected in 2/19 microsatellite instability and in 11/47 remaining cases (difference not significant). The survival of the microsatellite instability cases was significantly better. The presence of circulating cancer cells did not correlate with survival. CONCLUSION: It is possible that the microsatellite instability status, but not circulating cancer cells, constitutes a prognostic predictive factor in advanced gastric carcinoma. Confirmation of this hypothesis requires continuation of patient follow-up.

Adult↗

PubMed2001

Lack of prognostic influence of circulating tumor cells in peripheral blood of patients with colorectal cancer.

BACKGROUND AND AIMS: Circulating tumor cells in peripheral blood may be detected using high-sensitivity molecular techniques in several types of solid neoplasms, but their significance in colorectal cancer is controversial. The aim of this study was to assess the prognostic value of carcinoembryonic antigen (CEA) messenger RNA (mRNA) detection in peripheral blood samples from patients with colorectal cancer. METHODS: Peripheral vein blood samples from 95 consecutive patients with histologically confirmed colorectal carcinoma were obtained immediately before surgery to determine the presence of circulating tumor cells by use of a reverse-transcription polymerase chain reaction targeting CEA mRNA. Endpoints of the study were disease-free and overall survival. Results are referred to the whole series and, more importantly, to the 68 patients who underwent surgery for cure. RESULTS: After a median follow-up of 42 months, 19 of 68 patients (28%) operated on for cure had tumor relapse. In addition, 50 of 68 patients (73%) were alive. The probability of disease-free and overall survival was dependent on lymph node metastases and degree of differentiation, but not on the presence of circulating tumor cells (disease-free survival: relative risk, 1.00; 95% confidence interval [CI], 0.39-2.22, P = 0.99; overall survival: relative risk, 0.91, 95% CI, 0.34-2.43; P = 0.84). Similar results were obtained when all 95 patients with colorectal cancer were analyzed (disease-free survival: relative risk, 1.11; 95% CI, 0.63-1.95; P = 0.71; overall survival: relative risk, 1.21; 95% CI, 0.63-2.30, P = 0.55). CONCLUSIONS: Preoperative detection of blood circulating tumor cells by means of reverse-transcription polymerase chain reaction of CEA does not have prognostic significance in patients with colorectal cancer.

Aged↗

PubMed1996

Detection of ras gene mutations in perioperative peripheral blood with pancreatic adenocarcinoma.

Surgeons wish to know of any correlation between an operation and the incidence of metastasis. In perioperative periods, pancreatic cancer cells were identified by detecting mutant K-ras gene by two-step PCR and RFLP analysis in blood samples taken from peripheral blood. In no case was K-ras point mutation detected in blood before operation, although the mutant hand was observed in all cases at the time the lesion was resected. Surprisingly, in five of ten cases, positive bands were identified just after laparotomy, before we had reached the primary lesion. In almost all cases, mutant K-ras was detected until the fourteenth postoperative day. These findings suggest that cancer cells exist in the circulation, and have a potential for hematogenous metastasis during the perioperative period. In conclusion, surgical stress causes hematogenous dissemination of pancreatic cancer cells, and surgeons should employ the appropriate anti-metastasis therapy in the perioperative period.

Adenocarcinoma↗

PubMed1975

The significance of abnormal circulating cells in patients with Hodgkin's disease.

Buffy coat preparations of peripheral blood from two patients with Stage IVB Hodgkin's disease, 33 patients with Hodgkin's disease undergoing complete staging including laparotomy, 12 normal controls and eight patients with viral upper respiratory infections were examined for the presence of abnormal cells. Primitive histiocytic cells were seen in the two patients with IVB disease and in two patients with IIIB disease all of whom proved to be refractory to chemotherapy. Reed-Sternberg cells were seen in one IVB patient. Large cells with basophilic, agranular cytoplasm and moderately convoluted nuclei often with a perinuclear halo and nucleoli were detected in 30 of 35 patients with Hodgkin's disease, five of eight patients with viral infections but in none of 12 normal controls. Other abnormal large cells with round nuclei, occasional prominent nucleoli, grey to lightly eosinophilic cytoplasm and diffuse granulation were seen almost exclusively in Hodgkin's disease. There was no correlation between histology, stage, splenic involvement, skin test positivity and response to therapy with the presence or frequency of either of these cell types. This observation suggests that these cells are more likely to be 'reactive' rather than malignant. It is unusual for abnormal histiocytic cells to circulate in Hodgkin's disease and their presence may be a poor prognostic sign. The presence of other types of large atypical cells is not indicative of either haematogenous spread of Hodgkin's disease or of poor prognosis after treatment with radiotherapy alone.

Drug Therapy, Combination↗

PubMed1994

Detection of circulating tumor cells in men with localized prostate cancer.

PURPOSE: Using prostate-specific antigen (PSA) mRNA as a marker for prostatic epithelial cells, we have developed a sensitive technique that involves reverse transcription and polymerase chain reaction (RT-PCR) to detect circulating tumor cells in the peripheral blood of men with prostatic carcinoma (CaP). PATIENTS AND METHODS: A sensitive RT-PCR assay was used to evaluate the peripheral blood of 135 men with a history of CaP. Fourteen men with benign prostate disease, many of whom had elevated serum PSA levels, were used as a control group. RESULTS: All patients with benign prostate disease had a negative result in the RT-PCR assay. Of particular interest was a subgroup of 65 patients with clinically localized CaP evaluated before definitive local therapy. Five of these patients had detectable PSA mRNA by RT-PCR, suggesting circulating tumor cells. Within this group, systemic disease was detected by RT-PCR in some men with PSA levels less than 10 ng/mL and clinical stage B disease. Blood from men with hormone-refractory and progressive CaP demonstrated a higher frequency of PSA mRNA detectable by RT-PCR (10 of 20 patients). In contrast, none of seven patients with newly diagnosed metastatic prostate cancer and only one of seven patients with metastatic, hormone-responsive disease had blood that was positive for PSA mRNA by RT-PCR. CONCLUSION: Circulating tumor cells can be detected in the blood of a subset of patients with clinically localized CaP and a larger subset of patients with progressive metastatic disease.

Base Sequence↗