# CYTOSKELETAL HYBRIDOMA ANTIBODIES AS DIAGNOSTIC REAGENTS

> **NIH NIH R01** · UNIVERSITY OF WASHINGTON · 1993 · $189,302

## Abstract

Cell proliferation plays a critical role in the underlying events of
human neoplasia, as well as our understanding of tumor biology as it
relates to clinical course.  The development of slide-based methods to
assess cell proliferation is therefore of great interest with important
potential applications.  The overall goal of this project is to use
monoclonal antibodies and cDNA probes to cell proliferation related
markers (PCNA, H4 mRNA) and selected putative mediators of the loss of
proliferation control [(p53, retinoblastoma gene product (RB1), the B
chain of the platelet derived growth factor (PDGF-B)] on tissue sections
of human soft tissue sarcomas and melanomas.  A series of validation
experiments will first be performed to permit the application of the cell
proliferation markers to formalin and methacarn fixed, deparaffinized
human tumor specimens, and to test the hypothesis that alterations in the
distribution of proliferating cells within these tumors correlates with
histologic progression to malignancy and/or predicts clinical outcome.
It is then proposed to test the hypothesis that p53 overexpression, or
loss of RB1 expression, correlates with alterations in tumor
proliferation, as evidenced by quantitative and/or qualitative changes
in the expression of PCNA or H4 mRNA, as determined by immuno-
cytochemistry and in situ hybridization, in combination with cell-type
specific markers.  Quantification will be facilitated by employing image
analysis with a microcomputer-based quantitative microscopy system.  In
addition, analysis of expression by human tumors of the tumor suppressor
genes p53 and RB1, using immunocytochemistry, in situ hybridization,
Western and Northern blots, will be performed to test the hypothesis that
dysfunction of these proteins can define subsets of tumors with worse
clinical outcomes.  In addition, using these same tumors, the expression
of PDGF-B chain, a growth factor, will be investigated using a series of
monoclonal antibodies and cDNA probes in immunocytochemical as well as
Northern blot analyses, to test the hypothesis that PDGF-B chain
expression is correlated with paracrine stimulation of stromal cells in
melanomas, and with autocrine stimulation in the case of soft tissue
sarcomas.  New monoclonal antibodies to p53, RB1, and PDGF-B chain
protein which can be used on formalin fixed, paraffin embedded tissues
will also be developed to facilitate retrospective analyses.

## Key facts

- **NIH application ID:** 3173785
- **Project number:** 2R01CA036250-09A2
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **SAM.gov UEI:** HD1WMN6945W6
- **Principal Investigator:** ALLEN M. GOWN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $189,302
- **Award type:** 2
- **Project period:** 1984-03-01 → 1996-09-29

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3173785

## Citation

> US National Institutes of Health, RePORTER application 3173785, CYTOSKELETAL HYBRIDOMA ANTIBODIES AS DIAGNOSTIC REAGENTS (2R01CA036250-09A2). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3173785. Licensed CC0.

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