The long term objective of the proposed research is to precisely define the function of type II binding sites in normal and malignant cells. Nuclear type II sites bind methyl p-hydroxyphenyllactate (MeHPLA) and this compound appears to regulate normal and malignant cell growth through this binding interaction. Although type II binding sites may be closely coupled to DNA replication and cellular proliferation, the precise function of this nuclear protein and MeHPLA in normal and malignant cell regulation is unknown. The general goal of this proposal is to study type II gene structure and expression and subsequent effects on cellular proliferation. In order to accomplish this goal, the following aims are proposed: determine the biological origin of MeHPLA in mammalian cells, synthesize an affinity ligand, and purify nuclear type II sites from the rat or rabbit uterus. Type II sites will be extracted from rat or rabbit uterine nuclei and purified by chromatographic techniques for separation based on molecular size, charge and ligand binding specificity (affinity chromatography). Monoclonal and polyclonal antibodies will be generated against purified type II antigen and these immunological probes will be employed for qualitative and quantitative analysis of type II sites and the estrogen receptor. Type II antibodies and oligonucleotide probes will be used to screen cDNA libraries for the type II gene(s). The gene will be characterized and the cDNA utilized as a probe to study estrogen, MeHPLA, tamoxifen and carcinogen (dimethylbenz[a]anthracene) modulation of type II gene expression in normal (rat uterus) and malignant cells in vitro (human breast cancer cells) and in vivo (rat and mouse mammary tumor).

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA035480-08
Application #
3173049
Study Section
Reproductive Endocrinology Study Section (REN)
Project Start
1983-08-01
Project End
1994-07-31
Budget Start
1992-08-01
Budget End
1993-07-31
Support Year
8
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Type
Schools of Medicine
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
Markaverich, Barry M; Shoulars, Kevin; Rodriguez, Mary Ann (2011) Luteolin Regulation of Estrogen Signaling and Cell Cycle Pathway Genes in MCF-7 Human Breast Cancer Cells. Int J Biomed Sci 7:101-111
Markaverich, Barry M; Vijjeswarapu, Mary; Shoulars, Kevin et al. (2010) Luteolin and gefitinib regulation of EGF signaling pathway and cell cycle pathway genes in PC-3 human prostate cancer cells. J Steroid Biochem Mol Biol 122:219-31
Shoulars, Kevin; Rodriguez, Mary Ann; Thompson, Trellis et al. (2010) Regulation of cell cycle and RNA transcription genes identified by microarray analysis of PC-3 human prostate cancer cells treated with luteolin. J Steroid Biochem Mol Biol 118:41-50
Shoulars, Kevin; Rodriguez, Mary Ann; Thompson, Trellis et al. (2008) Regulation of the nitric oxide pathway genes by tetrahydrofurandiols: microarray analysis of MCF-7 human breast cancer cells. Cancer Lett 264:265-73
Markaverich, Barry M; Crowley, Jan; Rodriquez, Mary et al. (2007) Tetrahydrofurandiol stimulation of phospholipase A2, lipoxygenase, and cyclooxygenase gene expression and MCF-7 human breast cancer cell proliferation. Environ Health Perspect 115:1727-31
Markaverich, Barry M; Alejandro, Mary; Thompson, Trellis et al. (2007) Tetrahydrofurandiols (THF-diols), leukotoxindiols (LTX-diols), and endocrine disruption in rats. Environ Health Perspect 115:702-8
Markaverich, Barry M; Shoulars, Kevin; Alejandro, Mary-Ann (2006) Nuclear type II [3H]estradiol binding site ligands: inhibition of ER-positive and ER-negative cell proliferation and c-Myc and cyclin D1 gene expression. Steroids 71:865-74
Shoulars, Kevin; Rodriguez, Mary Ann; Crowley, Jan et al. (2006) Reconstitution of the type II [3H]estradiol binding site with recombinant histone H4. J Steroid Biochem Mol Biol 99:1-8
Shoulars, Kevin; Rodrigues, Mary Ann; Crowley, Jan R et al. (2005) Nuclear type II [3H]estradiol binding sites: a histone H3-H4 complex. J Steroid Biochem Mol Biol 96:19-30
Markaverich, Barry M; Crowley, Jan R; Alejandro, Mary A et al. (2005) Leukotoxin diols from ground corncob bedding disrupt estrous cyclicity in rats and stimulate MCF-7 breast cancer cell proliferation. Environ Health Perspect 113:1698-704

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