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Description
The purpose of this project was to identify proteins associated with the migration and invasion of non-transformed MCF10A mammary epithelial cells with ectopically expressed missense mutations in p53. Because of the prevalence of TP53 missense mutations in basal-like and triple-negative breast cancer tumors, understanding the effect of TP53 mutations on the phenotypic expression of human mammary epithelial cells may offer new therapeutic targets for those currently lacking in treatment options. As such, MCF10A mammary epithelial cells ectopically overexpressing structural mutations (G245S, H179R, R175H, Y163C, Y220C, and Y234C) and DNA-binding mutations (R248Q, R248W, R273C, and R273H) in the DNA-binding domain were selected for use in this project. Overexpression of p53 in the mutant cell lines was confirmed by western blot and q-PCR analysis targeting the V5 epitope tag present in the pLenti4 vector used to transduce TP53 into the mutant cell lines. Characterization of the invasion and migration phenotypes resulting from the overexpression of p53 in the mutant cell lines was achieved using transwell invasion and migration assays with Boyden chambers. Statistical analysis showed that three cell lines—DNA-contact mutants R248W and R273C and structural mutant Y220C—were consistently more migratory and invasive and demonstrated a relationship between the migration and invasion properties of the mutant cell lines. Two families of proteins were then explored: those involved in the Epithelial-Mesenchymal Transition (EMT) and matrix metalloproteinases (MMPs). Results of q-PCR and immunofluorescence analysis of epithelial marker E-cadherin and mesenchymal proteins Slug and Vimentin did not show a clear relationship between mRNA and protein expression levels with the migration and invasiveness phenotypes observed in the transwell studies. Results of western blotting, q-PCR, and zymography of MMP-2 and MMP-9 also did not show any consistent results indicating a definite relationship between MMPs and the overall invasiveness of the cells. Finally, two drugs were tested as possible treatments inhibiting invasiveness: ebselen and SBI-183. These drugs were tested on only the most invasive of the MCF10A p53 mutant cell lines (R248W, R273C, and Y220C). Results of invasion assay following 30 μM treatment with ebselen and SBI-183 showed that ebselen does not inhibit invasiveness; SBI-183, however, did inhibit invasiveness in all three cell lines tested. As such, SBI-183 will be an important compound to study in the future as a treatment that could potentially serve to benefit triple-negative or basal-like breast cancer patients who currently lack therapeutic treatment options.
ContributorsZhang, Kathie Q (Author) / LaBaer, Joshua (Thesis director) / Anderson, Karen (Committee member) / Gonzalez, Laura (Committee member) / Barrett, The Honors College (Contributor) / School of International Letters and Cultures (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
As the prevalence of childhood obesity in the United States rises, opportunities for children to be physically active become more vital. One opportunity for physical activity involves children walking to and from school. However, children that live in areas with a pedestrian-unfriendly built environment and a low degree of walkability are less likely to be physically active and more likely to be overweight. The purpose of this study was to study walking routes from schools in low-income neighborhoods in Southwestern United States to a local community center. Walking routes from the three study schools (South Mountain High School, Percy Julian Middle School, and Rose Linda Elementary School) were determined by distance, popularity, and the presence of a major thoroughfare. Segments and intersections, which formed the routes, were randomly selected from each school's buffer region. The walking routes as a whole, along with the segments and intersections, were audited and scored using built environment assessments tools: MAPS, PEQI and Walkability Checklist. These scores were utilized to develop interactive mapping tools to visualize the quality of the routes, segments and intersections and identify areas for improvement. Results showed that the routes from Percy Julian to the Kroc Center were, overall, rated higher than routes from the other two schools. The highest scoring route, from the seven routes studied, was route 2 from Percy Julian to the Kroc Center along Broadway Road. South Mountain High School was overall the worst starting point for walking to the Kroc Center as those three walking routes were graded as the least walkable. Possible areas for improvement include installing traffic calming features along major thoroughfares and reducing the perceived risk to pedestrian safety by beautifying the community by planting greenery. Future directions include studying the built environment in South Phoenix communities that surround the Kroc Center.
ContributorsZeien, Justin Lee (Author) / Buman, Matthew (Thesis director) / Hekler, Eric (Committee member) / Fellows, Brian (Committee member) / Barrett, The Honors College (Contributor) / School of Sustainability (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
Exercise is a great boon to both the health of individuals and the national as a whole. Exercise provides a wide range of significant and well established benefits to both physical and psychological health, with many benefits that are still being discovered. The effects of exercise on health are somewhat unique, as exercise is one of a limited number of ways to improve health that is not harm a reduction strategy, but instead increases health through direct benefit, rather than increasing health by decreasing damage and harm. Support is also given to the proposal that individuals are in best position to determine the intensity of exercise and to choose primary activities to participate in, in order to provide near maximum physical and psychological benefit, with the understanding that frequency of exercise is of the upmost importance for benefit. The accessibility of exercise and the tremendous health benefit of exercise, makes exercise a huge asset in reducing the exorbitant health care spending and improving mediocre health outcomes in this country; a reasonable goal as numerous countries have better health the United States, even though the United States spends the more than any other country on health.
ContributorsRael, Ashur Scott (Author) / Kashiwagi, Dean (Thesis director) / Kashiwagi, Jacob (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Film, Dance and Theatre (Contributor)
Created2015-05
Description
With higher than normal levels of cardiovascular disease, diabetes, obesity, and sexually transmitted diseases within the Deaf community, it is clear that the current level of medical care is failing. This can stem from many different issues, from the mistrust of doctors to the fact that scientific education in the Deaf community is introduced at a later time in comparison to the hearing population. Similarly, there is a distinct lack of Deaf and hard of hearing medical staff. All of this culminates in the fact that the system needs to change. The addition of more Deaf staff in the medical environment, more staff interpreters, and doctors that have experience with the Deaf community engenders the metamorphosis of this group from unhealthy to healthy. Extra exposure for doctors during clinical rotations to deaf patients as well as training in Deaf Culture will further increase the comfort level of Deaf community members utilizing health services. As a result, the overall state of medical care for the Deaf population will improve with these modifications to the current system.
ContributorsBrar, Serbnoor Singh (Author) / Quinn, Paul (Thesis director) / Howard, Pamela (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2015-05
Description
Sickle Cell Disease (SCD) is a prevalent genetic disease in Africa, and specifically in Kenya. The lack of available relevant disease education and screening mean that most don't understand the importance of getting testing and many children die before they can get prophylactic care. This project was designed to address the lack of knowledge with supplemental educational materials to be partnered with an engineering capstone project that provides a low cost diagnostic test.
ContributorsShawver, Jamie Christine (Author) / Caplan, Michael (Thesis director) / Snyder, Jan (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / Harrington Bioengineering Program (Contributor)
Created2014-05
Description
The focus of this project was to look at alternative treatments for endocrine resistant breast cancer (ERBC), which are breast cancers that have become resistant to hormone therapies such as Tamoxifen or aromatase inhibitors. The first part of this project involves investigating the relationship between histone de-acetylase inhibitor Vorinostat and Tamoxifen in MCF7 G11 cells, Tamoxifen resistant sub-clones, according to the PSOC Time grant. The second part involves targeting the androgen receptor (AR) in MCF7 sub-clones with AR antagonists, Bicalutamide and MDV3100, and investigating the possible usage of AR as a biomarker, due to over-expression of AR in ERBC, in accordance with the Mayo ASU Seed Grant.
The synergistic effects between Vorinostat and Tamoxifen observed through a phase II study on breast cancer patients resistant to hormone therapy may involve more than the modulation of ER-alpha to reverse Tamoxifen resistance in ERBC cells. RT-qPCR of genes expressed in Tamoxifen resistant cells, trefoil factor 1(TFF1) and v-myc avian myelocytomatosis viral oncogene homolog (MYC), were evaluated along with ESR1 and Diablo as a control. MYC was observed to have increased expression in the treated cells, whereas the other genes had a decrease in their expression levels after the cells were treated for 3 days with Vorinostat IC30 of 1 µM. As for targeting the AR, MCF7 Tamoxifen sensitive and resistant cells were not affected by the AR antagonists to determine an IC50. The cell viability for all MCF7 sub-clones only decreased for high concentrations of 5.56 µM - 50 µM in Bicalutamide and 16.67 µM – 50 µM of MDV1300. Furthermore, hormone depletion of MCF7 G11 Tamoxifen resistant sub-clones did not show a great response to DHT stimulation or the AR antagonists. In the RT-qPCR, the MCF7 G11 cells showed an increase in mRNA expression for ER, AR, and PR after 4 hours of treatment with estradiol. As for the DHT treatment, ER, AR, PR, and PSA had a minimal increase in the fold change, but the fold change in AR was less than in the estradiol treatment. The Mayo Clinic will investigate the possible usage of AR as a biomarker through immunohistochemistry.
The synergistic effects between Vorinostat and Tamoxifen observed through a phase II study on breast cancer patients resistant to hormone therapy may involve more than the modulation of ER-alpha to reverse Tamoxifen resistance in ERBC cells. RT-qPCR of genes expressed in Tamoxifen resistant cells, trefoil factor 1(TFF1) and v-myc avian myelocytomatosis viral oncogene homolog (MYC), were evaluated along with ESR1 and Diablo as a control. MYC was observed to have increased expression in the treated cells, whereas the other genes had a decrease in their expression levels after the cells were treated for 3 days with Vorinostat IC30 of 1 µM. As for targeting the AR, MCF7 Tamoxifen sensitive and resistant cells were not affected by the AR antagonists to determine an IC50. The cell viability for all MCF7 sub-clones only decreased for high concentrations of 5.56 µM - 50 µM in Bicalutamide and 16.67 µM – 50 µM of MDV1300. Furthermore, hormone depletion of MCF7 G11 Tamoxifen resistant sub-clones did not show a great response to DHT stimulation or the AR antagonists. In the RT-qPCR, the MCF7 G11 cells showed an increase in mRNA expression for ER, AR, and PR after 4 hours of treatment with estradiol. As for the DHT treatment, ER, AR, PR, and PSA had a minimal increase in the fold change, but the fold change in AR was less than in the estradiol treatment. The Mayo Clinic will investigate the possible usage of AR as a biomarker through immunohistochemistry.
ContributorsVorachitti, Merica (Author) / LaBaer, Joshua (Thesis director) / Anderson, Karen (Committee member) / Gonzalez, Laura (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05