The 92 kDa type IV collagenase (MMP-9) contributes to the spread of oral cancer and understanding how its expressions regulated could ultimately yield new agents to repress its expression and diminish tumor invasiveness. Although we previously identified multiple regulatory elements (AP-1, NF-kappaB, PEA3, Sp1) in the 670 base pair promoter, the limited number of these binding sites makes it unlikely that MMP-9 expression is solely the consequence of transactivation through these motifs. Indeed, emerging studies indicate a role for the chromatin environment (constituted by DNA wrapped around a histone core), in the regulation of gene expression. We show herein that Mtal, which promotes histone deacetylation, represses MMP-9 expression. Further, Mtal expression is undetectable in MMP-9-producing oral cancer.
In Specific Aim 1 using genomic foot printing and chromatin immunoprecipitation assays (Chip) we will identify transcription factor-bound cis elements and acetylated histones localized at these sites in the 670 base pair MMP-9 promoter targeted by Mtal to achieve MMP-9 repression. Biological suppressors of MMP-9 expression may also provide a tool for identifying regulatory elements in the chromatinized promoter. We show herein that the metastasis suppressor gene KiSS-1 attenuates MMP-9 transcription partly by reducing NF-kappaB binding to the chromatinized promoter. Nevertheless, the degree to which NF-kappaB binding is reduced can only partly account for the diminished transcription. Therefore, in Specific Aim 2, we will identify transactivated cis elements and acetylated histones localized at these sites in the MMP-9 promoter that mediate KiSS-1-dependent repression of MMP-9.Similarly, the MEK1 inhibitor PD098059 represses MMP-9 expression and in Specific Aim 3 we will determine if the transcriptional targets of this repressor are identical to, or distinct from, those of Mtal and KiSS-1. If we determine that the transcriptional targets differ, we will determine whether combining PD098059 and KiSS-1 or Mtal proves superior to individual modalities in reducing MMP-9 expression and oral cancer invasiveness. Since extra-chromosomal reporters were used in our previous studies of MMP-9 transcription, regulatory elements, that depend on the chromatin environment, may have escaped detection. Thus, to identify such novel cis elements in Specific Aim 4, we will employ DNase hypersensitivity, genomic foot printing and lambdagt11 library screening to identify additional transactivators/repressors of MMP-9 expression. Ultimately, the goal of these studies is to identify new transcriptional targets in the MMP-9 promoter that allow for therapeutic intervention to repress expression of this collagenase and oral cancer invasiveness.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
2R01DE010845-09A1
Application #
6574169
Study Section
Special Emphasis Panel (ZRG1-OBM-1 (01))
Program Officer
Shirazi, Yasaman
Project Start
1994-02-01
Project End
2007-11-30
Budget Start
2003-02-01
Budget End
2003-11-30
Support Year
9
Fiscal Year
2003
Total Cost
$264,250
Indirect Cost
Name
University of Texas MD Anderson Cancer Center
Department
Biology
Type
Other Domestic Higher Education
DUNS #
800772139
City
Houston
State
TX
Country
United States
Zip Code
77030
Ishiguro, T; Avila, H; Lin, S-Y et al. (2010) Gene trapping identifies chloride channel 4 as a novel inducer of colon cancer cell migration, invasion and metastases. Br J Cancer 102:774-82
Yang, Lin; Zhang, Li; Wu, Qiuyu et al. (2008) Unbiased screening for transcriptional targets of ZKSCAN3 identifies integrin beta 4 and vascular endothelial growth factor as downstream targets. J Biol Chem 283:35295-304
Yang, Lin; Hamilton, Stanley R; Sood, Anil et al. (2008) The previously undescribed ZKSCAN3 (ZNF306) is a novel ""driver"" of colorectal cancer progression. Cancer Res 68:4321-30
Nair, Rajesh R; Avila, Hector; Ma, Xujun et al. (2008) A novel high-throughput screening system identifies a small molecule repressive for matrix metalloproteinase-9 expression. Mol Pharmacol 73:919-29
Wang, H; Yan, C; Asangani, I et al. (2007) Identification of an histone H3 acetylated/K4-methylated-bound intragenic enhancer regulatory for urokinase receptor expression. Oncogene 26:2058-70
Yan, Chunhong; Boyd, Douglas D (2007) Regulation of matrix metalloproteinase gene expression. J Cell Physiol 211:19-26
Nair, Rajesh R; Solway, Julian; Boyd, Douglas D (2006) Expression cloning identifies transgelin (SM22) as a novel repressor of 92-kDa type IV collagenase (MMP-9) expression. J Biol Chem 281:26424-36
Yan, Chunhong; Boyd, Douglas D (2006) Histone H3 acetylation and H3 K4 methylation define distinct chromatin regions permissive for transgene expression. Mol Cell Biol 26:6357-71
Yan, Chunhong; Jamaluddin, Md S; Aggarwal, Bharat et al. (2005) Gene expression profiling identifies activating transcription factor 3 as a novel contributor to the proapoptotic effect of curcumin. Mol Cancer Ther 4:233-41
Yan, Chunhong; Lu, Dan; Hai, Tsonwin et al. (2005) Activating transcription factor 3, a stress sensor, activates p53 by blocking its ubiquitination. EMBO J 24:2425-35

Showing the most recent 10 out of 39 publications