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PubMed2004

In vivo flow cytometry: a new method for enumerating circulating cancer cells.

The fate of circulating tumor cells is an important determinant of their ability to form distant metastasis. Here, we demonstrate the use of in vivo flow cytometry as a powerful new method for detecting quantitatively circulating cancer cells. We specifically examine the circulation kinetics of two prostate cancer cell lines with different metastatic potential in mice and rats. We find that the cell line and the host environment affect the circulation kinetics of prostate cancer cells, with the intrinsic cell line properties determining the initial rate of cell depletion from the circulation and the host affecting cell circulation at later time points.

Animals↗
PubMedDate not supplied

Cavernous transformation of the portal vein secondary to tumor thrombosis of hepatocellular carcinoma: spiral CT visualization of the collateral vessels.

BACKGROUND: We investigated the constituting collateral vessels in cavernous transformation of the portal vein (CTPV) caused by tumor thrombosis of hepatocellular carcinoma (HCC) by using contrast-enhanced spiral computed tomographic (CT) examination. METHODS: Fifty-four histopathologically proven HCC patients with tumor thrombosis-induced CTPV were retrospectively included and assigned to cirrhosis negative (n = 31) and positive (n = 23) groups. Another 15 cirrhotic patients with portal hypertension but no HCC and CTPV were used for comparison. Standardized dual-phase contrast-enhanced spiral CT was performed for all patients. CT appearances of the collateral vessels of CTPV were observed, and their visualization rates were analyzed. RESULTS: Biliary (cystic and paracholedochal veins) and gastric (left and right gastric veins) branches of the portal vein were the most frequently visualized collateral vessels of CTPV. There was a marked difference in CT visualization rates for biliary branches between patients with and without CTPV (83-94% vs. 0). No difference existed in visualization rates for gastric branches across the three groups (77-87% for left gastric, 58-61% for right gastric vein). CONCLUSIONS: Biliary and gastric branches of the portal vein are the major collateral vessels of CTPV. The intergroup differences in CT visualization rates may provide clues to the roles that they might play in the hemodynamic adaptation process of CTPV.

Adult↗
PubMed1996

Induction of tumour cell shedding into effluent venous blood breast cancer surgery.

Surgeons have long been concerned that cancer may be disseminated by shedding of tumour cells into the bloodstream during surgery. Early claims that cancer operations induced an increase in the number of tumour cells shed into the circulation were subsequently discredited, and the issue has remained unresolved. We used immunocytochemistry for cytokeratins to detect tumour cells in effluent blood from breast carcinomas in 18 patients undergoing surgery. Tumour cells were detectable in 6/18 patients during surgery, in only one patient before operation and in none post-operatively (P = 0.025). Circulating cells were associated with vascular invasion within the primary tumour (P = 0.032). No cytokeratin-positive cells were found in blood from ten normal volunteers or four patients undergoing surgery for other breast conditions. These results confirm that cancer surgery in humans results in an increase in the shedding of tumour cells into the circulation. The implications for prognosis and practice should be determined by larger prospective studies.

Breast Neoplasms↗
PubMed1976

Similarities of surface characteristics of neoplastic well-differentiated lymphocytes from solid tissues and from peripheral blood.

Cells from spleens and lymph nodes infiltrated with neoplastic well-differentiated lymphocytes (WDLT) were investigated for the presence of receptors for unsensitized sheep red blood cells, complement (IgM EAC), and antigen-antibody complexes, as well as for the presence of surface immunoglobulins. In addition, the cells were examined by scanning electron microscopy (SEM). The surface markers and ultrastructure of WDLT cells were compared with those of cells obtained from peripheral blood of patients with chronic lymphocytic leukemia (CLL). Like the CLL populations, most WDLT samples showed a low percentage of unsensitized sheep red blood cell-binding cells and a concomitantly high percentage of IgM EAC-binding cells. Furthermore, a substantial number of WDLT cells in several instances showed a lower binding affinity for IgM EAC than normal lymphocytes, as demonstrated by the poor attachment of the IgM EAC reagent to the WDLT cells in suspension and to frozen tissue sections. Weak IgM EAC binding capacity was demonstrated in the CLL cells as well. WDLT and CLL cells bound the IgG EA complex only when certain sensitive techniques were applied (pelleting). A variable percentage of cells with surface immunoglobulins was found among WDLT and DLL populations. However, accurate counting of surface immunoglobulin-bearing neoplastic cells was very difficult in most cases due to the extremely faint surface fluorescence as compared to normal. SEM did not reveal specific changes in WDLT or CLL cells that would permit their identification as neoplastic lymphocytes. No major differences were found between WDLT and CLL cells and no correlation could be established between SEM appearances and surface markers. As previously observed with other cell populations, the conditions under which the cells were manipulated before fixation (in particular the environmental temperature) seemed to play a major role in the final SEM appearance of both WDLT and CLL cells. The results of our studies indicate that WDLT cells represent a population of B-cells whose surface properties are very similar to those of circulating CLL cells. The poor capacity of WDLT cells and CLL cells to bind IGM EAC or to stain with fluorescein-conjugated antihuman immunoglobulin suggests low density or poor affinity of the surface receptors. In this regard, these neoplastic cells are different from those of follicular lymphomas, also of B-cell origin.

Adult↗
PubMed2000

The detection of prostate cells by the reverse transcription-polymerase chain reaction in the circulation of patients undergoing transurethral resection of the prostate.

OBJECTIVE: To determine whether prostate cells are disseminated into the circulation of patients after transurethral resection of the prostate (TURP), as assessed by a reverse transcription-polymerase chain reaction (RT-PCR) assay for prostate specific antigen (PSA) mRNA. PATIENTS AND METHODS: Fifty-one patients, comprising 34 with benign prostatic hyperplasia (BPH) and 17 with prostate cancer who were undergoing routine TURP, had blood samples taken before and 30 min after surgery. The blood mononuclear cell layer was isolated by density-gradient centrifugation and total RNA extracted. Complementary DNA was synthesized by RT of the RNA. The target PSA sequence was amplified by PCR with specific PSA primers and the product detected on agarose-gel electrophoresis with ethidium bromide staining. RESULTS: Five patients (all with prostate cancer) were positive on PSA RT-PCR before surgery and remained positive after TURP. Of the remaining 46 patients, five (11%; three with BPH and two with cancer) changed from a negative to a positive result after TURP. CONCLUSION: In a proportion of patients TURP causes the dissemination of prostate cells into the circulation which are then detectable by RT-PCR of PSA mRNA.

Aged↗
PubMed2000

Significant detection of circulating cancer cells in the blood by reverse transcriptase-polymerase chain reaction during colorectal cancer resection.

OBJECTIVE: To analyze the clinical value of reverse transcriptase-polymerase chain reaction (RT-PCR) recognition of mRNA coding for carcinoembryonic antigen (CEA) and cytokeratin 20 in blood obtained from patients with colorectal carcinoma. SUMMARY BACKGROUND DATA: RT-PCR has been applied to identify very small numbers of tumor cells. Molecular detection is thought to provide useful information for the clinical management of perioperative prophylaxis of tumor cell implantation or postoperative adjuvant therapy regimens. METHODS: From 52 patients with colorectal cancer, peripheral blood specimens were obtained before and after surgical manipulation; also, a specimen of mesenteric venous blood draining the colorectal tumor was obtained just before tumor resection. Using cDNA primers specific for CEA and cytokeratin 20, RT-PCR was performed to detect tumor cells. Subsequently, the 52 patients were divided into two groups, a group positive for both CEA and cytokeratin 20 and a group negative for CEA, cytokeratin 20, or both. RESULTS: On the basis of 450 days of follow-up data, the PCR-positive group had a significantly shorter overall survival than the PCR-negative group only with the mesenteric venous blood specimens. Multivariate analysis indicated that detection of the simultaneous presence of CEA and cytokeratin 20 mRNA in mesenteric venous blood is a potent prognostic factor independent of the traditional pathologic parameters. Of the eight peripheral blood specimens found to be PCR-positive, five showed a change of PCR from negative to positive during surgery, and liver metastases developed 11 months later in one of these five patients. CONCLUSIONS: Molecular detection of both CEA and cytokeratin 20 mRNA in mesenteric venous blood may be of prognostic value for patients with colorectal carcinoma. Molecular detection in the peripheral blood at surgery suggests that hematogenic tumor cell dissemination is a common and early event and that surgical manipulation enhances this release of tumor cells into the circulation.

Adult↗
PubMed1996

Role of Kupffer cells in arresting circulating tumor cells and controlling metastatic growth in the liver.

Metastasis to the liver is a common event in clinical oncology. Blood-borne tumor cells (TCs) arriving to the liver sinusoids run into a special vascular bed. The lining of liver sinusoids is shared by Kupffer cells (KCs) and endothelial cells. KCs, liver-fixed macrophages, are responsible for detection and removal of "non-self" particles. To investigate their role in arresting blood-borne TCs and controlling tumor growth, we injected a syngeneic colon carcinoma cell line into a mesenteric vein of two groups of rats; one group was without Kupffer cells and the other normal controls. We removed the liver of these animals at different time intervals and performed immunohistochemical analysis with monoclonal antibodies (MoAbs) against our tumor cell line, three macrophage subpopulations, natural killer cells, and B and T lymphocytes. Additionally, we showed in vitro spontaneous cytotoxicity of KCs against our tumor cell line. Results suggest that KCs play a relevant role in arresting circulating TCs at the liver sinusoid, although it is limited to a small number of malignant cells. They also seem to play a major role in clearing neoplastic cells from the liver parenchyma, in controlling tumor growth in the very early stages of metastatic development, and in modulating the host immune response to cancer cells.

Animals↗