Optimization of prostate cancer cell detection using multiplex tyramide signal amplification. Issue 4 (2nd November 2018)

• Record Type: Journal Article
• Authors: Roy, Sounak; Axelrod, Haley D.; Valkenburg, Kenneth C.; Amend, Sarah; Pienta, Kenneth J.
• Is Part Of: Journal of cellular biochemistry. Volume 120:Issue 4(2019)
• Page Start: 4804
• Page End: 4812
• Publication Date: 2018-11-02
• Subjects: detection -- immunofluorescence -- metastasis -- prostate cancer -- tumor microarray -- tyramide; Cytochemistry -- Periodicals; 572
• DOI: 10.1002/jcb.28016
• Languages: English
• ISSNs: 0730-2312

Abstract

Abstract: Approximately 29 000 men die of prostate cancer (PCa) each year in the United States, and 90% to 100% of them are due to incurable bone metastasis. It is difficult to determine (1) when PCa disseminates in the natural history of the disease; (2) where cancer cell disseminates before becoming overt metastatic lesions; and (3) which tumors are aggressive and which are indolent. Tumor tissue and liquid (blood and bone marrow) biopsies provide important information to answer these questions, but significant limitations exist for immunostaining strategies that assess protein expression in these tissues. Classic immunohistochemistry (IHC) assays can typically assess expression of one or two proteins per tissue section. We have developed a novel immunofluorescence staining protocol to detect a panel of seven proteins on PCa tissue from primary tumor biopsies and metastatic lesion autopsy tissue, as well as cancer cells from liquid biopsies. We used a tyramide‐based system to amplify the true signal and optimized the protocol to reduce background signal, thereby boosting the signal‐to‐noise ratio. Any protein‐specific antibody in this protocol can be exchanged for a different validated antibody. This protocol therefore, represents a highly informative and flexible assay that can be used to provide important information about cancer tissue for the purpose of improving detection, diagnosis, and treatment. Abstract : Ninety to hundred percent of men who die from prostate cancer (PCa) die due to incurable bone metastasis and it is difficult to determine (1) when PCa disseminates in the natural history of the disease; (2) where cancer cell disseminate before becoming overt metastatic lesions; and (3) which tumors are aggressive and which are indolent. We have developed a novel immunofluorescence staining protocol to detect a panel of seven proteins on PCa tissue from primary tumor biopsies, metastatic lesion autopsy tissue, and cancer cells from liquid biopsies to improve PCa cell detection. This protocol is a highly informative and flexible assay in which protein‐specific antibodies can be easily substituted for different validated antibodies and can be used to provide important information about cancer tissue for the purpose of improving detection, diagnosis, and treatment.