Prostate cancer: Flow cytometric methods for detection of bone marrow micrometastases

Hussain, Maha; KuKuruga, Mark; Biggar, Sandra; Sakr, Wael; Cummings, Glenn; Ensley, John F.

doi:10.1002/(sici)1097-0320(19960315)26:1<40::aid-cyto6>3.0.co;2-k

Cited by 15 publications

(4 citation statements)

References 20 publications

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“…This cell line was de- rived from a grade IV prostatic adenocarcinoma [16]. The PC-3 cell line has been shown, to be positive for cytokeratin 8 and 18 but negative for PSMA [10,18]. LNCaP cells were purchased from ATCC.…”

Section: Methodsmentioning

confidence: 99%

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Method for identifying prostate cells in semen using flow cytometry

Barren¹,

Holmes²,

Boynton³

et al. 1998

Prostate

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BACKGROUND Prostate‐specific antigen (PSA) cannot differentiate benign prostatic hyperplasia (BPH), from prostatitis, or prostate cancer in the range of 4.0–10 ng/ml. An accurate cytologic or histologic assessment is necessary to confirm the proper diagnosis. The nature of a biopsy tends to make it a selective test not frequently repeated. We are reporting a technique employing semen as a source for the differential diagnosis of prostate epithelial cells. METHODS Eleven vasectomized and nonvasectomized prostate cancer patients provided semen samples (stage T1 to T2). Two patients provided repeat samples. In addition, 15 vasectomized or nonvasectomized individuals without evidence of disease provided semen samples. Three million cells fixed with 50% ethanol were stained by an antibody (7E11.C5) to prostate‐specific membrane antigen (PSMA), Hybritech Antibody (399) to PSA, and cytokeratin 8 and 18. In addition to the antibodies described, a DNA stain To‐Pro 3 was used to identify 2n‐4n DNA containing cells. A dual laser, Becton Dickinson FACSCaliber cytometer, was used to analyze the samples. RESULTS All semen specimens contained diploid, cytokeratin 18–positive epithelial cells regardless of disease status. A clear difference between prostate cancer and normal prostate cell samples was observed using staining with 7E11.C5. The ratio of prostatic cells in the total epithelial cell population (PSMA:cytokeratin ratios) was calculated for each specimen. A retrospective study of sixteen semen samples from 11 prostate cancer patients had a mean PSMA:cytokeratin ratio of 0.57, whereas the samples from 15 patients without evidence of cancer had a mean PSMA:cytokeratin ratio of 0.11. This difference was significant. PSA staining was variable and inconsistent. CONCLUSIONS This report demonstrates that human semen contains prostate cells that can be characterized and used in the clinical diagnosis of prostate cancer. Prostate 36:181–188, 1998. © 1998 Wiley‐Liss, Inc.

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Section: Methodsmentioning

confidence: 99%

“…LNCaP is a cell line developed from a secondary tumor extracted from the lymph node of a hormone-refractory prostate cancer patient [19]. The LNCaP cell line has been shown to be positive for both cytokeratin 8 and 18 and PSMA [10,18]. Cells were grown in monolayer to confluence and harvested with 0.02% ethylenediaminetetraacetic acid in PBS.…”

Section: Methodsmentioning

confidence: 99%

Method for identifying prostate cells in semen using flow cytometry

Barren¹,

Holmes²,

Boynton³

et al. 1998

Prostate

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“…These include DNA ploidy [12], differentiation markers [13], and cytokeratin-positive/CD45 À cells [14]. The same principle has been used in a Food and Drug Administration-licensed test, which first immunomagnetically preseparates circulating cells positive for an epithelial cell marker before detecting CD45 À cytokeratin þ cells by automated fluorescence microscopy [15,16].…”

mentioning

confidence: 99%

Rare-Event Analysis in Flow Cytometry

Donnenberg

2007

Clinics in Laboratory Medicine

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“…Cell sorting is easy to do and enables a high throughput of samples, quantification of results, and isolation of subpopulations of cells. The feasibility of using FACS assays for detecting micrometastases was reported in several cancers [12,13,20,23]. Compared with normal individuals there are significantly more CECs identified by FACS analysis in patients with prostate cancer.…”

Section: Introductionmentioning

confidence: 99%

Evaluation and significance of circulating epithelial cells in patients with hormone‐refractory prostate cancer

García¹,

Rosenberg²,

Weinberg

et al. 2007

BJU International

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The urological oncology section is relatively long this month, and this reflects the many high‐quality manuscripts we receive. When you consider our relatively high rejection rate, you will understand just how many papers on this topic are submitted. The high quality of oncology papers is clear in this month’s section. You will also notice that all but one of them are on prostate cancer, and the reason for this is similar to that mentioned above, as this topic is, as might be expected, the most commonly submitted in this section. However, I am only too happy to reassure readers, and those primarily interested in other types of urological cancer, that the imbalance in this month’s section is not a permanent fixture. OBJECTIVE To determine the feasibility of using flow cytometry fluorescence‐activated cell sorting (FACS) analysis for detecting circulating epithelial cells (CECs) in patients with hormone‐refractory prostate cancer (HRPC), and to determine whether CECs can be used to predict survival in these patients. PATIENTS AND METHODS Several prognostic models that include routinely used clinical and laboratory variables for predicting survival in men with HRPC have been reported; the presence of CECs measured by reverse transcriptase‐polymerase chain reaction for prostate‐specific antigen (PSA) in patients with HRPC is an independent prognostic factor for survival. CECs detected by FACS analysis correlate with advanced stage and poor survival outcome. A retrospective study was conducted to assess the presence of CECs by FACS analysis in metastatic HRPC patients initiating systemic chemotherapy with a taxane‐based regimen. The association between clinical variables previously described and the presence of CECs along with the effect of the magnitude of CECs on survival was calculated, in 41 patients with HRPC, all of whom had peripheral blood collected for FACS analysis. RESULTS Except for four patients, all those with metastatic HRPC had detectable CECs. Among these patients, the number of CECs/mL was correlated with age, serum PSA level and serum alkaline phosphatase (ALP). Higher serum levels of PSA and ALP predicted a poor survival outcome. Similarly, patients with ≤1.8 CECs/mL had a significantly longer survival than those with more CECs/mL (P = 0.02). With a median follow‐up of 15.4 months, the median overall survival for all patients was 18.4 months. CONCLUSIONS The presence of more CECs in patients with metastatic HRPC was associated with a poorer survival outcome; levels of ≥1.8 CECs/mL were associated with a shorter survival in patients with metastatic HRPC.

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Prostate cancer: Flow cytometric methods for detection of bone marrow micrometastases

Cited by 15 publications

References 20 publications

Method for identifying prostate cells in semen using flow cytometry

Method for identifying prostate cells in semen using flow cytometry

Rare-Event Analysis in Flow Cytometry

Evaluation and significance of circulating epithelial cells in patients with hormone‐refractory prostate cancer

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