Frequent Detection and Immunophenotyping of Prostate-Derived Cell Clusters in the Peripheral Blood of Prostate Cancer Patients

2004 ◽  
Vol 19 (2) ◽  
pp. 93-99 ◽  
Author(s):  
H. Schmidt ◽  
G. De Angelis ◽  
O. Bettendorf ◽  
E. Eltze ◽  
A. Semjonow ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Protein Expression ◽  
Cancer Cells ◽  
Cancer Patients ◽  
Peripheral Blood ◽  
Preoperative Staging ◽  
Specific Antigen ◽  
Cell Clusters ◽  
Cancer Aggressiveness ◽  
Associated Proteins

Background Recent scientific studies have failed to determine parameters for the assessment of prostate cancer aggressiveness. The present study deals with the detection of blood-borne cancer cells based on polymerase chain reaction (PCR) and cell enrichment methods. The contradictory results reported in the literature have called into question the clinical usefulness of this diagnostic method in the preoperative staging of clinically localized prostate cancer. Methods We established a combined method of density gradient centrifugation and immunomagnetic separation using epithelium-specific antibodies, i.e. cytokeratins, to isolate prostate-derived circulating cells from the peripheral blood of patients with prostate cancer. Isolated cells were characterized by DNA staining and immunocytochemistry using antibodies for the detection of prostate-specific antigen (PSA), proliferation-associated proteins (MIB-1, H1 and H3) and apoptosis-associated proteins (M30, c-FasR). Results We applied these methods to 68 prostate cancer patients and were able to isolate cell clusters in 98%. Immunophenotypic and morphological characterization of PSA-positive prostate-derived cell clusters found in the peripheral blood of prostate cancer patients showed two main populations: 1) in 35% of the investigated prostate cancer patients we detected rounded cell aggregates of probable cancer cells expressing proliferation-associated proteins and lacking apoptosis-associated protein expression; 2) in all cases there was a high frequency of circulating dysmorphic cell clusters positive for apoptosis-associated protein expression. Conclusion Our results demonstrate the existence of at least two different types of blood-borne prostate-derived circulating cell clusters. Of these, only the less frequent, round, small cell clusters harbor features that are probably necessary for the cells to survive for metastatic spread.

Blood-Borne Cancer Cells – Quo Vadis?

2000 ◽  
Vol 15 (1) ◽  
pp. 111-113 ◽  
Author(s):  
B. Brandt ◽  
H. Schmitt ◽  
J.C. Feldner ◽  
R.J. Lellé ◽  
A. Semjonow ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cancer Patients ◽  
Peripheral Blood ◽  
Cancer Metastasis ◽  
Specific Antigen ◽  
Clinical Staging ◽  
Breast Cancer Patients ◽  
Cell Clusters ◽  
The Impact

The detection of blood-borne cancer cells may help in clinical staging and further understanding of cancer metastasis. We developed a cytokeratin-based immunomagnetic method to isolate epithelium-derived cells from the circulating blood of patients. The number of cell clusters positive for cytokeratin/prostate-specific antigen (PSA) from the peripheral blood of prostate cancer patients and cytokeratin/p185c-erbB-2 from the peripheral blood of breast cancer patients has been related to stage of the disease. Breast cancer patients who presented cytokeratin/p185c-erbB-2-positive cell clusters showed a decrease in such cells under adriamycin adjuvant therapy with Further molecular characterization by a highly sensitive microsatellite multiplex-PCR enabled reproducible detection of microsatellite alterations. The impact of these individually targeted results may contribute to an individual diagnostic and therapeutic strategy.

scholarly journals Glycidamide Promotes the Growth and Migratory Ability of Prostate Cancer Cells by Changing the Protein Expression of Cell Cycle Regulators and Epithelial-to-Mesenchymal Transition (EMT)-Associated Proteins with Prognostic Relevance

2019 ◽  
Vol 20 (9) ◽  
pp. 2199
Author(s):  
Titus Ime Ekanem ◽  
Chi-Chen Huang ◽  
Ming-Heng Wu ◽  
Ding-Yen Lin ◽  
Wen-Fu T. Lai ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Cycle ◽  
Protein Expression ◽  
Cancer Cells ◽  
Cancer Patients ◽  
Epithelial To Mesenchymal Transition ◽  
Gene Signature ◽  
Prostate Cancer Cells ◽  
Mesenchymal Transition ◽  
Prognostic Relevance

Acrylamide (AA) and glycidamide (GA) can be produced in carbohydrate-rich food when heated at a high temperature, which can induce a malignant transformation. It has been demonstrated that GA is more mutagenic than AA. It has been shown that the proliferation rate of some cancer cells are increased by treatment with GA; however, the exact genes that are induced by GA in most cancer cells are not clear. In the present study, we demonstrated that GA promotes the growth of prostate cancer cells through induced protein expression of the cell cycle regulator. In addition, we also found that GA promoted the migratory ability of prostate cancer cells through induced epithelial-to-mesenchymal transition (EMT)-associated protein expression. In order to understand the potential prognostic relevance of GA-mediated regulators of the cell cycle and EMT, we present a three-gene signature to evaluate the prognosis of prostate cancer patients. Further investigations suggested that the three-gene signature (CDK4, TWIST1 and SNAI2) predicted the chances of survival better than any of the three genes alone for the first time. In conclusion, we suggested that the three-gene signature model can act as marker of GA exposure. Hence, this multi-gene panel may serve as a promising outcome predictor and potential therapeutic target in prostate cancer patients.

Ex vivo propagation and characterization of circulating tumor cell clusters from late stage prostate cancer patients.

2016 ◽  
Vol 34 (2_suppl) ◽  
pp. 314-314 ◽  
Author(s):  
James Lim ◽  
Charles J. Ryan ◽  
Bruce Adams ◽  
Jeff Hough ◽  
Tianna Chow
Keyword(s):  
Prostate Cancer ◽  
T Cells ◽  
Dendritic Cells ◽  
Tumor Cell ◽  
Cancer Patients ◽  
Peripheral Blood ◽  
Biomarker Discovery ◽  
Ex Vivo ◽  
Circulating Tumor Cell ◽  
Cell Clusters

314 Background: Cells shed by a tumor into the bloodstream upon metastasis present an opportunity to study the properties of these cells for biomarker discovery, drug development, and personalized therapy. Enrichment and expansion of circulating tumor cells (CTCs) in culture enables molecular characterization not readily available for rare cells. Here, we demonstrate the successful identification and culture of rare circulating tumor cell clusters from peripheral blood of metastatic castration resistant prostate cancer patients (mCRPC). Methods: The approach to isolate CTC clusters relies on nucleated cell enrichment via Ficoll-based centrifugation from 15 mL of peripheral blood. A collagen-based cell-binding substrate was used to capture cell clusters with preferential affinity for collagen (i.e. epithelial-like cells). Hypoxic and pressurized culturing conditions were used to maintain and propagate adhered CTC clusters. After 7 days in culture, immunofluorescence imaging, qPCR and/or mRNAseq were performed on CTC clusters that had formed colonies. Results: Blood samples processed within 3 hours from time of draw yielded a 25% success rate (4/16) for identifying PSMA+/EpCAM+/CD45- CTC colonies. Morphologically distinct CTC colonies cultivated at 1% oxygen and increased hydrostatic pressure (2 PSI) were positive for PSMA and EpCAM staining as observed via confocal microscopy, with individual colonies ranging from approximately 50 to 1000 in cell number. Colonies consistently revealed white blood cell contaminants (T-cells and dendritic cells, 10 to 30% of colony) that did not appear to perturb colony growth. CTC colonies were collected for qPCR analysis and mRNAseq via laser-capture microdissection and/or micropipetting. Genomic analysis revealed differential gene expression patterns between CTC colonies cultured from the same patient sample, with a subset expressing neuroendocrine like signatures (AURKA), stem cell markers (SOX2, OCT4), and immuno-oncology relevant markers (PD-L1, CTLA4). Conclusions: In conclusion, we show that viable CTCs are amenable to ex vivo culture, providing both functional and molecular insights into a sub-population of CTCs with propagating potential. The presence of T-cells and dendritic cells on CTC colonies warrants further investigation, and may provide unique insights into immune-tumor interactions. The culturing platform is currently being evaluated as a research tool for biomarker discovery and as a clinical tool for disease monitoring and treatment decision-making.

Cytokeratin 19 Expression by Reverse Transcriptase-Polymerase Chain Reaction in the Peripheral Blood of Prostate Cancer Patients

2005 ◽  
Vol 91 (3) ◽  
pp. 248-252 ◽  
Author(s):  
Marcos Tobias-Machado ◽  
Fernando Fonseca ◽  
Ana Paula Fantinato ◽  
Israel Bendit ◽  
Marcelo Langer Wroclawski ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Cancer Patients ◽  
Peripheral Blood ◽  
Specific Antigen ◽  
Cytokeratin 19 ◽  
Chain Reaction ◽  
Peripheral Blood Mononuclear ◽  
Polymerase Chain

Aims and background Sensitive reverse transcriptase-polymerase chain reaction-based techniques for detection of epithelial antigen expression, such as cytokeratin 19, in the peripheral blood mononuclear fraction of prostate cancer patients may allow the detection of tumor progression at A molecular level. Methods We studied cytokeratin 19 expression by reverse transcriptase-polymerase chain reaction in peripheral blood mononuclear cell samples of 10 control men and serially in 44 patients with prostate cancer every three months for 18 months. Results None of the 10 normal control men expressed cytokeratin 19 in their peripheral blood mononuclear fraction. In the patients, cytokeratin 19 positivity at entry was not associated with age, Gleason score, clinical stage, prostate-specific antigen or alkaline phosphatase. Interestingly, having at least one positive cytokeratin 19 result during follow-up correlated significantly with time to prostate-specific antigen progression (P = 0.049), especially in the subgroup of metastatic patients (P = 0.032). Conclusions We conclude that cytokeratin 19 expression by reverse transcriptase-polymerase chain reaction in the peripheral blood mononuclear cell fraction of prostate cancer patients correlates with time to prostate-specific antigen progression. Further studies are needed to confirm these findings.

scholarly journals α-Particle-induced DNA damage tracks in peripheral blood mononuclear cells of [223Ra]RaCl2-treated prostate cancer patients

2021 ◽  
Author(s):  
S. Schumann ◽  
U. Eberlein ◽  
C. Lapa ◽  
J. Müller ◽  
S. Serfling ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Dna Damage ◽  
Cancer Patients ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Absorbed Dose ◽  
Peripheral Blood Mononuclear ◽  
Absorbed Doses ◽  
Blood Mononuclear Cells

Abstract Purpose One therapy option for prostate cancer patients with bone metastases is the use of [223Ra]RaCl2. The α-emitter 223Ra creates DNA damage tracks along α-particle trajectories (α-tracks) in exposed cells that can be revealed by immunofluorescent staining of γ-H2AX+53BP1 DNA double-strand break markers. We investigated the time- and absorbed dose-dependency of the number of α-tracks in peripheral blood mononuclear cells (PBMCs) of patients undergoing their first therapy with [223Ra]RaCl2. Methods Multiple blood samples from nine prostate cancer patients were collected before and after administration of [223Ra]RaCl2, up to 4 weeks after treatment. γ-H2AX- and 53BP1-positive α-tracks were microscopically quantified in isolated and immuno-stained PBMCs. Results The absorbed doses to the blood were less than 6 mGy up to 4 h after administration and maximally 16 mGy in total. Up to 4 h after administration, the α-track frequency was significantly increased relative to baseline and correlated with the absorbed dose to the blood in the dose range < 3 mGy. In most of the late samples (24 h – 4 weeks after administration), the α-track frequency remained elevated. Conclusion The γ-H2AX+53BP1 assay is a potent method for detection of α-particle-induced DNA damages during treatment with or after accidental incorporation of radionuclides even at low absorbed doses. It may serve as a biomarker discriminating α- from β-emitters based on damage geometry.

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