RNA-binding proteins to assess gene expression states of co-cultivated cells in response to tumor cells

Luiz OF Penalva¹^,3 , Michael D Burdick¹^,4 , Simon M Lin² , Hedwig Sutterluety¹^,5 and Jack D Keene¹

¹Department of Molecular Genetics and Microbiology, Center for RNA biology, 414 Jones Building, Research Drive, Duke University Medical Center, Durham, North Carolina 27710 USA

²Duke Bioinformatics Shared Resource, Duke University Medical Center, Durham, North Carolina 27710 USA

³Children's Cancer Research Institute. UTHSCSA. Mail code 7784 7703 Floyd Curl Dr. San Antonio, TX 78229-3900 USA

⁴Bayer Corporation 85 T.W. Alexander Drive Research Triangle Park NC 27709 USA

⁵Institut fur Krebsforschung Abt. Fur Angewandte und Experimentelle Onkologie Borschkegasse 8a A-1090 Wien Austria

author email corresponding author email

Molecular Cancer 2004, 3:24doi:10.1186/1476-4598-3-24


Published:	7 September 2004

Abstract

Background

Tumors and complex tissues consist of mixtures of communicating cells that differ significantly in their gene expression status. In order to understand how different cell types influence one another's gene expression, it will be necessary to monitor the mRNA profiles of each cell type independently and to dissect the mechanisms that regulate their gene expression outcomes.

Results

In order to approach these questions, we have used RNA-binding proteins such as ELAV/Hu, poly (A) binding protein (PABP) and cap-binding protein (eIF-4E) as reporters of gene expression. Here we demonstrate that the epitope-tagged RNA binding protein, PABP, expressed separately in tumor cells and endothelial cells can be used to discriminate their respective mRNA targets from mixtures of these cells without significant mRNA reassortment or exchange. Moreover, using this approach we identify a set of endothelial genes that respond to the presence of co-cultured breast tumor cells.

Conclusion

RNA-binding proteins can be used as reporters to elucidate components of operational mRNA networks and operons involved in regulating cell-type specific gene expression in tissues and tumors.