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Updated: Aug 19, 2026
The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers
Published on: January 22, 2013
Use of reverse phase protein microarrays and reference standard development for molecular network analysis of
Katherine M Sheehan1, Valerie S Calvert, Elaine W Kay
1United States Food and Drug Administration (FDA)-NCI Clinical Proteomics Program, Laboratory of Pathology, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, MD, USA.
Abstract:
Cancer can be defined as a deregulation or hyperactivity in the ongoing network of intracellular and extracellular signaling events. Reverse phase protein microarray technology may offer a new opportunity to measure and profile these signaling pathways, providing data on post-translational phosphorylation events not obtainable by gene microarray analysis. Treatment of ovarian epithelial carcinoma almost always takes place in a metastatic setting since unfortunately the disease is often not detected until later stages. Thus, in addition to elucidation of the molecular network within a tumor specimen, critical questions are to what extent do signaling changes occur upon metastasis and are there common pathway elements that arise in the metastatic microenvironment. For individualized combinatorial therapy, ideal therapeutic selection based on proteomic mapping of phosphorylation end points may require evaluation of the patient's metastatic tissue. Extending these findings to the bedside will require the development of optimized protocols and reference standards. We have developed a reference standard based on a mixture of phosphorylated peptides to begin to address this challenge.
Insights
Reverse phase protein microarray technology enables profiling of cancer signaling pathways. A new reference standard using phosphorylated peptides aids in analyzing metastatic ovarian cancer for targeted therapies.
Area of Science:
- Oncology
- Proteomics
- Molecular Biology
Background:
- Cancer is characterized by dysregulated intracellular and extracellular signaling.
- Ovarian epithelial carcinoma is often diagnosed at a metastatic stage.
- Understanding signaling in metastasis is crucial for effective treatment.
Purpose of the Study:
- To explore the utility of reverse phase protein microarray technology for profiling cancer signaling pathways.
- To investigate signaling changes in metastatic ovarian cancer.
- To develop a reference standard for proteomic analysis in cancer research.
Main Methods:
- Utilized reverse phase protein microarray technology to analyze signaling pathways.
- Examined post-translational phosphorylation events.
- Developed a reference standard comprising a mixture of phosphorylated peptides.
Main Results:
- Reverse phase protein microarray technology provides data on phosphorylation events.
- Identified potential common pathway elements in the metastatic microenvironment.
- Successfully developed a phosphorylated peptide mixture as a reference standard.
Conclusions:
- Proteomic mapping of phosphorylation endpoints can inform individualized cancer therapy.
- Evaluating metastatic tissue is essential for selecting optimal treatments.
- The developed reference standard is a step towards clinical application of proteomic analysis in oncology.
