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Detection of circulating breast cancer cells by reverse transcriptase polymerase chain reaction (RT-PCR).

The confounding problem in treatment of breast cancer is the metastasis of breast tumour. Reverse transcriptase polymerase chain reaction (RT-PCR) has been recently used in the detection of circulating breast cancer cells. This review reports on the development of this assay as well as its advantages and disadvantages. We feel that cytokeratin 20 and beta -human chorionic gonadotropin (hCG) mRNA are the best markers for the detection of circulating breast cancer cells. We suggest that the multiple RNA marker RT-PCR assay can help to increase both sensitivity and specificity of detection, and that quantitative RT-PCR assay is more effective than the qualitative assay in the detection of circulating breast cancer cells.

Biomarkers, Tumor↗

Metastatic carcinoma with carcinocythemia mimicking leukemia.

Carcinoma metastatic to the bone marrow may present with peripheral blood and bone marrow changes and a clinical picture difficult to distinguish from leukemia. It is important to make an accurate diagnosis since the management is different. We have described a patient with circulating primitive cells and diffuse marrow involvement by similar cells. The pathologic process may be identified without cell markers by careful attention to the pattern of distribution, as well as the cytologic features of the neoplastic cells in the bone marrow biopsy.

Biopsy↗

Extrahepatic collateral supply of hepatocellular carcinoma by the intercostal arteries.

PURPOSE: To evaluate computed tomographic (CT) and angiographic findings of extrahepatic collateral supply of hepatocellular carcinoma (HCC) by the intercostal artery (ICA) and the efficacy of transcatheter arterial chemoembolization (TACE) in the ICA. MATERIALS AND METHODS: The CT and angiographic findings of 30 ICA collateral supplies of HCC in 19 patients were evaluated. TACE of the ICA collaterals was performed in 10 patients. The clinical outcomes and complications were evaluated. RESULTS: ICA collaterals were found at the first to 18th sessions of TACE of HCC. The CT findings were: large HCC (mean diameter, 10.3 cm), subcapsular location (94.7%), defect in iodized oil retention or progression of HCC at subcapsular region (31.6%), HCC abutting the abdominal wall in a broad area with or without abdominal wall invasion (63.2%), hypertrophied ICA (31.6%), and branching collateral vessels coursing from the abdominal wall to the HCC (26.3%). On angiograms, all ICA collaterals originated from the right side at levels of T8 (6.7%), T9 (30.0%), T10 (46.7%), or T11 (16.7%). Twelve sessions of TACE of the ICA collaterals were performed in 10 patients. Follow-up angiography was performed in six patients and showed persistent obliteration in one, recanalization in three, and progression in two. Complications were shoulder pain (n = 2), itching sensation (n = 1), erythema of skin (n = 1), and skin necrosis (n = 1). CONCLUSION: ICA collateral supply of HCC usually occurs in advanced HCC or after multiple sessions of TACE. When there are suggestive CT findings, ICA collaterals should be sought when TACE is performed in the management of HCC.

Abdominal Wall↗

Surgical approach to renal cell carcinoma extending into the right atrium through the inferior vena cava.

Successful management of a patient with an intracardiac tumor thrombus of renal carcinoma is described. This case and a few others in the literature have led us to consider the clinical signs of cavo-atrial obstruction, frequently silent and unspecific; the diagnostic methodology, especially based upon CAT scan and cavography, and the type of surgery and surgical technique called for, especially as regards the approach and the possible use of extracorporeal circulation (ECC).

Adenocarcinoma↗

Detection of circulating albumin-mRNA by RT-PCR does not indicate metastasizing hepatocellular carcinoma.

Hepatocellular carcinomas (HCC) frequently recur after partial liver resection or orthotopic liver transplantation, possibly because of the presence of a small number of hepatoma cells in the peripheral blood. Detection of circulating HCC cells might improve therapeutic options and could predict disease recurrence resulting from a metastasizing disease. In the present study, human albumin-mRNA was detected by RT-PCR in the peripheral blood of patients with hepatocellular carcinoma. Circulating albumin-specific PCR products were detected in each patient with HCC, but also in healthy volunteers. It is concluded that albumin-mRNA is not specific to circulating hepatoma cells and therefore does not indicate metastasizing disease.

Carcinoma, Hepatocellular↗

The detection of circulating breast cancer cells in peripheral blood by reverse transcriptase-polymerase chain reaction.

Some circulating cancer cells in the blood play a central role in the metastatic process and may have a major influence on patient progress. Their numbers can be very small and techniques for their detection need to be both sensitive and specific. Polymerase chain reaction (PCR) has been successfully used to detect small numbers of tumor cells in cancer. We used a reverse transcriptase-polymerase chain reaction (RT-PCR) to detect circulating breast cancer cells in venous blood samples before operations and assessed cytokeratin-19 (CK-19) and cytokeratin-20 (CK-20) as target mRNA markers in the blood of healthy donors (n=6) and breast cancer patients (n=30) with American Joint Committee on Cancer stages 0 to IIIa. CK-19 mRNA was expressed in all blood samples of healthy donors and patients. But CK-20 was the only mRNA marker not detected in the blood from healthy donors. Seven of 30 (23%) venous blood isolates of breast cancer patients yielded a CK-20 mRNA with positive results. There was no correlating CK-20 mRNA expression with stage and axillary lymph node status. In conclusion, CK-19 showed no diagnostic value as a mRNA marker in the detection of circulating cancer cells by RT-PCR assay because this was expressed in the blood of healthy donors. CK-20 mRNA was an useful marker to detect circulating cancer cells in breast cancers.

Breast Neoplasms↗

Utility of tests for circulating melanoma cells in identifying patients who develop recurrent melanoma.

Prospective studies were carried out on 186 patients with AJCC stage I (13), II (76) and III (97 patients) melanoma before and after surgical removal of their tumour. The goal was to determine if PCR tests on blood samples for MART-1 and tyrosinase were predictive of recurrence of melanoma in a 2-year follow-up period. (PCR assays for MUC-18, p97 and gp100 were positive in blood samples from normal subjects and excluded from the study.) PCR tests for MART-1 and tyrosinase were most commonly positive in the first 3 months following surgical removal of melanoma, and three tests over this period gave maximum sensitivity in the identification of patients who subsequently developed recurrences. Positive tests for the first time in the second year of follow-up had similar predictive power. The tests identified 68.5% of patients who developed recurrences in the 2-year follow-up period. Assays for MART-1 were mainly positive in patients with locoregional recurrences, whereas tyrosinase was detected in blood samples from patients with both locoregional and disseminated recurrences. (Positivity rate for tyrosinase in 48 patients with disseminated melanoma was 60.4% compared to 14.6% for MART-1.) Tests for MART-1 and tyrosinase were strongly predictive of disease-free survival (DFS) and were more powerful predictors of DFS than lymph node status or thickness of the primary melanoma. (Hazard ratios by Cox analysis were 2.97 in patients with disseminated recurrences and 2.93 in those with locoregional recurrences.) These results indicate that PCR tests for MART-1 and tyrosinase are powerful prognostic indicators, but their practical utility for selecting patients for adjuvant therapy is limited by the high false-negative rate of approximately 30%. Intermittent shedding of melanoma cells into the circulation would appear to be the most likely explanation for the latter.

Antigens, Neoplasm↗

Effects of whole body hyperthermia (41.8 degrees C) on the frequency of tumor cells in the peripheral blood of patients with advanced malignancies.

PURPOSE: Combining heat with antineoplastic drugs has produced evidence of antitumor synergism. An increasing number of trials are investigating whole body hyperthermia (WBH) in combination with chemotherapy in patients with advanced malignancies. Here we investigated whether the hyperdynamic state of the circulation as a consequence of WBH may stimulate dissemination of malignant cells. EXPERIMENTAL DESIGN: WBH in combination with chemotherapy was administered by a radiant heat device to 20 consecutive patients with advanced epithelial malignancies. One WBH session lasted for approximately 4 h (90 min heating time, 60 min plateau at 41.8 degrees C, and 60-80 min cooling). Peripheral blood was drawn before WBH treatment (baseline), at the end of the plateau (1 h), and 24 h and 48 h thereafter. After removal of leukocytes using anti-CD45 magnetic beads, circulating tumor cells were detected immunocytochemically using the monoclonal antibody A45-B/B3, which binds to a common epitope present on various cytokeratins. RESULTS: The method used to detect tumor cells in the peripheral blood proved to be specific and very sensitive (detection limit 1 tumor cell per 1.7 x 10(5) peripheral blood mononuclear cell). Before WBH, 6 of 20 patients had cyto-keratin-positive cells in their blood. A treatment-induced increase in the number of circulating tumor cells became statistically significant at 24 h after WBH (P = 0.043) and was detected in a total of 9 patients, 5 of whom had no detectable malignant cells at baseline. There was no evidence of a correlation between an increase in the number of circulating tumor cells and increased metastasis frequency. CONCLUSIONS: Our findings suggest that WBH might induce a temporary release of tumor cells into the circulation, but this spread appears to be clinically not significant in patients with advanced malignancies.

Adult↗