Specific detection of prostate cancer cells in urine by multiplex immunofluorescence cytology

Fujita, Kazutoshi; Pavlovich, Christian P.; Netto, George J.; Konishi, Yuko; Isaacs, William B.; Ali, Syed; De Marzo, Angelo; Meeker, Alan K.

doi:10.1016/j.humpath.2009.01.004

« Previous Next »Human Pathology
Volume 40, Issue 7 , Pages 924-933, July 2009

Specific detection of prostate cancer cells in urine by multiplex immunofluorescence cytology☆

Kazutoshi Fujita, MD
- Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Department of Urology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
Christian P. Pavlovich, MD
- Department of Urology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
George J. Netto, MD
- Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Department of Urology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Department of Oncology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
Yuko Konishi, BS
- Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
William B. Isaacs, PhD
- Department of Urology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Department of Oncology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
Syed Ali, MBBS
- Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
Angelo De Marzo, MD, PhD
- Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Department of Urology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Department of Oncology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
Alan K. Meeker, PhD
- Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Department of Urology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA
- Corresponding author. Division of Genitourinary Pathology, Department of Pathology, The Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA.

Received 10 November 2008; received in revised form 7 January 2009; accepted 9 January 2009. published online 15 April 2009.

Summary

Prostate cancer biomarkers are enriched in urine after prostatic manipulation, suggesting that whole cells might also be detectable for diagnosis. We tested multiplex staining of urinary sediments as a minimally invasive method to detect prostate cancer. Urine samples were collected from 35 men who had prostatic massage (attentive digital rectal examination) in a urology clinic and from 15 control men without urologic disease and without massage, for a total of 50 specimens (27 cancer-positive cases and 23 cancer-negative cases). LNCaP prostate cancer cells spiked into urine were used for initial marker optimization. Urine sediments were cytospun onto glass slides and stained. Multiplex urine cytology was compared with conventional urine cytology for cancer detection; anti–α-methylacyl-CoA racemase antibody was used as a marker of prostate cancer cells, anti-Nkx3.1 as a marker of prostate epithelial cells, anti-nucleolin as a marker of nucleoli, and 4′-6-diamidino-2-phenylindole to highlight nuclei. Prostate cancer cells were successfully visualized by combined staining for α-methylacyl-CoA racemase, Nkx3.1, and nucleolin. Of the 25 informative cases with biopsy-proven prostate cancer, 9 were diagnosed as suspicious or positive by multiplex immunofluorescence urine cytology, but only 4 were similarly judged by conventional cytology. All cases without cancer were read as negative by both methods. The multiplex cytology sensitivity for cancer detection in informative cases was 36% (9/25), and specificity was 100% (8/8). In conclusion, we have successfully achieved multiple staining for α-methylacyl-CoA racemase, Nkx3.1, nucleolin, and 4′-6-diamidino-2-phenylindole to detect prostate cancer cells in urine. Further refinements in marker selection and technique may increase sensitivity and applicability for prostate cancer diagnosis.

Keywords: Prostate cancer, Urine cytology, Biomarkers, Diagnosis, Multiplex staining

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☆ This work was supported by the following research funding mechanisms: NIH/NIDDK grant 1K23DK071262 (C. P. Pavlovich), Department of Defense grant W81XWH-05-1-0167 (C. P. Pavlovich), and a Patrick C. Walsh Foundation grant (A. K. Meeker).

PII: S0046-8177(09)00011-2

doi:10.1016/j.humpath.2009.01.004

« Previous Next »Human Pathology
Volume 40, Issue 7 , Pages 924-933, July 2009

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