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A high-content, multiplexed screen in human breast cancer cells identifies profilin-1 inducers with anti-migratory activities

Joy, ME and Vollmer, LL and Hulkower, K and Stern, AM and Peterson, CK and Boltz, RC and Roy, P and Vogt, A (2014) A high-content, multiplexed screen in human breast cancer cells identifies profilin-1 inducers with anti-migratory activities. PLoS ONE, 9 (2).

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Abstract

Profilin-1 (Pfn-1) is a ubiquitously expressed actin-binding protein that is essential for normal cell proliferation and migration. In breast cancer and several other adenocarcinomas, Pfn-1 expression is downregulated when compared to normal tissues. Previous studies from our laboratory have shown that genetically modulating Pfn-1 expression significantly impacts proliferation, migration, and invasion of breast cancer cells in vitro, and mammary tumor growth, dissemination, and metastatic colonization in vivo. Therefore, small molecules that can modulate Pfn-1 expression could have therapeutic potential in the treatment of metastatic breast cancer. The overall goal of this study was to perform a multiplexed phenotypic screen to identify compounds that inhibit cell motility through upregulation of Pfn-1. Screening of a test cassette of 1280 compounds with known biological activities on an Oris™ Pro 384 cell migration platform identified several agents that increased Pfn-1 expression greater than two-fold over vehicle controls and exerted anti-migratory effects in the absence of overt cytotoxicity in MDA-MB-231 human breast cancer cells. Concentration-response confirmation and orthogonal follow-up assays identified two bona fide inducers of Pfn-1, purvalanol and tyrphostin A9, that confirmed in single-cell motility assays and Western blot analyses. SiRNA-mediated knockdown of Pfn-1 abrogated the inhibitory effect of tyrphostin A9 on cell migration, suggesting Pfn-1 is mechanistically linked to tyrphostin A9's anti-migratory activity. The data illustrate the utility of the high-content cell motility assay to discover novel targeted anti-migratory agents by integrating functional phenotypic analyses with target-specific readouts in a single assay platform. © 2014 Joy et al.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Joy, MEmej29@pitt.eduMEJ29
Vollmer, LLllv4@pitt.eduLLV4
Hulkower, K
Stern, AMSTERNAM@pitt.eduSTERNAM
Peterson, CK
Boltz, RCrcb56@pitt.eduRCB56
Roy, Ppar19@pitt.eduPAR19
Vogt, Aavogt@pitt.eduAVOGT
Contributors:
ContributionContributors NameEmailPitt UsernameORCID
EditorWeaver, Alissa M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Centers: Other Centers, Institutes, or Units > Drug Discovery Institute
Other Centers, Institutes, or Units > Magee-Women's Research Institute
Date: 10 February 2014
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 9
Number: 2
DOI or Unique Handle: 10.1371/journal.pone.0088350
Schools and Programs: School of Medicine > Computational and Systems Biology
School of Medicine > Pathology
Swanson School of Engineering > Bioengineering
Refereed: Yes
Date Deposited: 18 Jun 2014 20:29
Last Modified: 22 Jan 2019 16:55
URI: http://d-scholarship.pitt.edu/id/eprint/21905

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