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Paraben and Breast Cancer: a Stromal Link

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Paraben and Breast Cancer: a Stromal Link University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 5-2020 Paraben and Breast Cancer: A Stromal Link Emily Hager University of Tennessee, [email protected] Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Recommended Citation Hager, Emily, "Paraben and Breast Cancer: A Stromal Link. " PhD diss., University of Tennessee, 2020. https://trace.tennessee.edu/utk_graddiss/5915 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by Emily Hager entitled "Paraben and Breast Cancer: A Stromal Link." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the equirr ements for the degree of Doctor of Philosophy, with a major in Nutritional Sciences. Ling Zhao, Major Professor We have read this dissertation and recommend its acceptance: Jay Whelan, Guoxen Chen, Hwa-Chain Wang Accepted for the Council: Dixie L. Thompson Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) Paraben and Breast Cancer: A Stromal Link A Dissertation Presented for the Doctor of Philosophy Degree The University of Tennessee, Knoxville Emily Nicole Hager May 2020 ACKNOWLEDGMENTS I am greatly honored and humbled to have pursued my Ph.D. at the University of Tennessee, Knoxville (UTK). As I grew up here in Knoxville, UTK has always been esteemed throughout my life, and it brings me great honor and joy to now be able to matriculate from such a distinguished university. I must first thank Dr. Ling Zhao for her continued support during my journey here at UTK. I have endured many life struggles whilst a student under Dr. Zhao’s tutelage, and she never gave up on me. There were times that she had to be stern with me, and I am so grateful for the strength she maintained to push me through these last six years. I have learned a great deal from Dr. Zhao, not only in terms of science and research but of life. I will never forget the compassion she has expressed towards me. From the depths of my heart, thank you. I would also like to thank my committee members for always emulating such wisdom and support. Dr. Jay Whelan has supported me in times of enormous despair. At one point in my doctorate pursuit, I was extremely lost, and with Dr. Whelan’s guidance, I was able to pull through and sustain my pursuit. Thank you, Dr. Whelan, for committing so much to the students that sit under your lead. You truly are the greatest example of leadership that I have observed. Dr. Guoxun Chen has brought so much wisdom and knowledge of the scientific field as well as brought joy to the laboratory on a daily basis. Every day he would walk through with a smile on his face, and sometimes that is all we need to see when life seems difficult. Lastly, Dr. Robert Wang has broadened my understanding of cancer biology more than any text or lecture could by explaining the underpinnings of published literature. It is an honor to sit under such established, wise, and encouraging advisors. Lastly, I want to recognize my husband and four kids that have never stopped believing in me. My husband speaks encouragement to me daily, even when I doubt myself. He also lovingly corrects me when I am being prideful or stubborn. My beautiful children have sat at my desk for hours, watched me work on my computer at night and on weekends, and have spoken encouraging words to me when I wanted to quit. Little Juliet has said so ii many times, “I want to go to the lav.” My father, aunt, in-laws, church, and family friends have been absolutely necessary for my completion of this degree. It has taken a village, and when I lost my previous husband so many people stepped in to help me watch my son. Lastly, I want to recognize Dr. Whelan and Dr. Zhao as they have done everything within their power to enable my continuance in this degree despite my world crumbling around me. I have had some of my most traumatize life experiences and some of my most amazing life experiences while pursuing this degree, and I am truly grateful to be surrounded by such a supportive, loving family. Thank you! iii ABSTRACT Breast cancer, surpassing lung cancer with the highest estimated new cases, is closely associated with obesity. Cancer research continues to divulge the complexity of the disease including the recognition that stromal cells must cooperate with the cancerous epithelium for cancer growth and metastasis. Parabens are a class of esters of 4-hydroxybenzoic acid that is ubiquitous in the environment, and act as anti-microbial agents that sustain the shelf-life of personal care products, pharmaceuticals, food, etc. Parabens are classified as endocrine disrupting chemicals, are estrogenic, and bioaccumulate within adipose tissue. Activation of fibroblasts is caused by tissue damage leading to the release of inflammatory cytokines to stimulate wound healing. Low dose (0.02, 0.1, and 1 μM) methyl-, propyl, and butylparaben (MP, PP, and BP) treatment onto murine 3T3-L1 and human primary breast (BRF) fibroblasts led to an inflammatory response by the upregulation of inflammatory cytokines including interleukin-1β (IL-1β) and cyclooxygenase-2 (COX2) via activation of the NF-κB pathway (Chapter III). MP, PP, and BP also upregulated matrix metalloproteinase-9 (MMP9), indicating potential influence in metastasis. Paraben-treated fibroblasts also increased the mRNA expression of CYP19A1 otherwise known as aromatase, which influences the conversion of androgen to estradiol. Secretions from paraben-treated fibroblasts also obtained the ability to enhance proliferation of estrogen receptor positive (ER+) breast cancer cells (MCF7). To evaluate the contribution of adipocytes, 3T3-L1 and BRF adipocytes were treated with MP, PP, and BP and induced upregulation IL-1β, COX2, MMP9, and CYP19A1 (Chapter IV). The secretions from paraben-treated adipocytes also enhanced the proliferation of MCF7 cells. In conclusion, we have shown that environmental achievable low doses of parabens enhance the inflammatory response of stromal fibroblasts and adipocytes, and the secretions produced from paraben exposure enhance the proliferation of breast cancer cells. Our results support the existing evidence that paraben exposure may lead to increased risks of breast cancer and suggest the stromal link between parabens and breast cancer. Further iv studies are warranted to confirm the effects of parabens on breast stromal cells and carcinogenesis in vivo and to establish the role of NF-B pathway. v TABLE OF CONTENTS Chapter 1 : Introduction .................................................................................................... 1 1.1. References ............................................................................................................... 5 1.2. Appendix ................................................................................................................. 8 Chapter 2 : Literature Review ......................................................................................... 10 2.1. Introduction ........................................................................................................... 11 2.2. Parabens, Endocrine Disruptors, and Breast Cancer ............................................ 12 2.2.1. Human Paraben Exposure ......................................................................... 12 2.2.2. Parabens are EDCs .................................................................................... 13 2.2.3. Parabens and Breast Cancer ...................................................................... 14 2.3. Estrogenic Effects of Parabens ............................................................................. 15 2.3.1. Estrogenicity ............................................................................................. 15 2.3.2. Mammary Tissue Development ................................................................ 16 2.3.3. Menstrual Cycles ...................................................................................... 17 2.4. Mechanisms of Action by Parabens ...................................................................... 18 2.4.1. Estrogen Receptor ..................................................................................... 18 2.4.2. Progesterone Receptor .............................................................................. 19 2.4.3. Human Epidermal Growth Factor Receptor ............................................. 20 2.4.4. Androgen and Glucocorticoid Receptor ................................................... 20 2.4.5. G-Protein Coupled Receptor ..................................................................... 21 2.5. Parabens Effect on Breast Cancer Progression ..................................................... 21 2.5.1. Mimic Estrogen in Inducing ER+ Breast Cancer Gene Expression ......... 22 2.5.2. Induce Genome Instability ........................................................................ 23 2.5.3. Evoke Breast Cancer Proliferation............................................................ 26 2.6. Parabens Effect on Breast Cancer Metastasis
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