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University of Colorado Cancer Center

University of Colorado Cancer Center, A National Cancer Institute-designated Comprehensive Cancer Center
 

Twila Jackson, PhD

Preceptor


Specialty:

Basic Reproductive Sciences

Location:

University of Colorado Anschutz Medical Campus, Aurora

Positions Available: 1

Objective:

Estrogen (E2) binds to and activates the estrogen receptor to regulate both normal and cancerous growth of uterine, breast and ovarian tissues. Estrogen receptors (ER) function as ligand-dependent (estrogen is the ligand) transcription factors which control target gene expression.

Upon estrogen binding, receptors localize to the nucleus, bind estrogen response elements in gene promoters, and regulate transcription. Recently, estrogen receptors have been shown to localize to the plasma membrane, associate with membrane proteins, and function as signal transduction molecules—this activity is termed extranuclear, nongenomic or nonclassical action.

The goal of this project is to elucidate the mechanisms by which E2 and ER initiate rapid cytoplasmic signaling. We will test the hypothesis that E2-activated ER interact with c-Src and PI3 kinases to initiate the signaling cascades that control uterine cancer cell growth.

Description:  

Endometrial (uterine) cancer cells expressing a mutant version of the ER that is solely cytosolic will be treated +/- estrogen and harvested under non-denaturing (native protein folding) conditions. Subsequent co-immunoprecipitation (co-IP) will be used to determine if ER can interact with the signaling kinases, c-Src and PI3 kinase. Co-IP uses antibodies to ER to precipitate ER and associated proteins out of the cell lysate. The immunoprecipitated proteins will be resolved by polyacrylamide gel electrophoresis and Western blotting will be used to determine if c-Src and PI3 kinase interact with ER.

The contribution of these kinases to uterine cancer cell growth will be determined using kinase inhibitors in conjuction with proliferation assays.

The student will learn:

  1.  basic tissue culture techniques

  2. protein extraction, immunoprecipitation, and immunoblotting

  3. densitometric quantitation and basic statistical analyses, and

  4. how to keep a lab notebook, interpret data and write an abstract. My goal is for the student to produce publishable data and leave the lab with potential co-authorship on a peer-reviewed journal article.