The Role of Vitamin D in Breast Cancer

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The effects of radiation on vitamin D actions in normal mammary cells, by Natalia L. Martinez

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  • Lobules- milk producing glandsDucts-tube that carries milk to the nipple.
  • Cellular proliferation-reproduction of new cellsDifferentiation-specialize; to change form one form to anotherApoptosis –necessary death of cells
  • The Role of Vitamin D in Breast Cancer

    1. 1. The Role of Vitamin D in Breast Cancer : The effects of radiation on vitamin D actions in normal mammary cells Department of Chemistry, The College of New Rochelle Natalia L. Martinez Mentor: Elvira Longordo University at Albany Summer Research Program GEN*NY*Sis Center for Excellence in Cancer Genomics JoEllen Welsh and Erika LaPorta
    2. 2. Breast Cancer • Breast Cancer is the uncontrolled growth of breast cells, in the lobules or the ducts Normal Cancer
    3. 3. Cellular Origin of Cancer
    4. 4. What Causes Cancer? Intrinsic Factors • Heredity • Diet • Hormones Extrinsic Factors • Chemicals • Radiation
    5. 5. DNA Damage • Chemicals and Radiation can damage DNA • Cells can sense and repair damage – p53- acts in the damage response mechanism – tumor suppressor • There is now a focus on identifying additional agents that enhance DNA repair •http://www.haemophilia.org.nz/DBimages/FCK/Image/dna.jpg
    6. 6. The link between Cancer & vitamin D • Recent research has shown a link between vitamin D and the prevention and treatment of Breast Cancer • Role of Vitamin D –Regulates Calcium and Phosphorus levels in the blood –Prevention of Rickets
    7. 7. How is Vitamin D obtained? • Food Sources – Fatty Fish – Eggs – Fortified Foods • Sunlight Exposure – UVB light
    8. 8. From Vitamin D to 1α,25(OH)2D3 7-dehydrocholesterol found in the skin cholecalciferol Or vitamin D3. forming 25- hydroxycholecalciferolProducing the active form of vitamin D3, 1,25-dihydroxycholecalciferol.
    9. 9. UV Stimulated Synthesis of Calcitriol Biochemistry Fifth Ed. Berg, Tymoczo and Stryer. W.H. Freeman, 2002.
    10. 10. The vitamin D receptor (VDR) • It is found in the nucleus of different tissues through the body • VDR is a nuclear hormone receptor • VDR is the major mediator of 1,25D actions
    11. 11. 1,25D & VDR When the active form of vitamin D binds to its nuclear receptor VDR it regulates gene expression which influences cellular proliferation, differentiation and apoptosis
    12. 12. In My Project: • Study the effects of radiation on vitamin D actions in normal human mammary cells (HME) Hypothesis: • DNA damage caused by ionizing radiation will up- regulate p53 and VDR expression Western Blot • Protection of the cells can be achieved by the ligand binding of 1,25D3 to VDR in damaged cells Crystal violet assay & immunoflurescent microscopy
    13. 13. Investigated the effects of radiation on vitamin D receptor (VDR) expression
    14. 14. Procedure • Cell Culture -HME cells (9 plates) • Ionizing Radiation DOSES OF EXPOSURE Incubation Period. Sham-0KV-0mA; 1min. 3 plates –analyzed after 1hr. 2Gy- 130KV- 2mA; 1min. 3 plates- analyzed after 3hr. 8Gy- 130KV- 7.7mA; 1 min 3 plates- analyzed after 6hr. http://www.instapstudycenter.net/images/faxitron.GIF
    15. 15. Detection of p53 and VDR by Western Blot Lamin A/C Lamin A/C 8Gy 2Gy Sham. 8Gy. 2Gy Sham Sham 2Gy 8Gy p53 VDR 6hrs. 3hrs. 1hr.
    16. 16. Use immunofluorescence microscopy to monitor expression and sub-cellular localization of VDR after radiation exposure in the presence and absence of 1,25D Detection of VDR by Confocal Microscopy www.ion.ucl.ac.uk/.../neuropath/techniques.htm
    17. 17. Results 1,25DEtOH
    18. 18. Results – 1hr
    19. 19. Results – 3hr
    20. 20. Examine whether treatment with 1,25D alters HME cell response to radiation.
    21. 21. Procedure • Plated cells in 6 well plates • Treated with 100nM Vitamin D – 48hr. Prior to radiation exposure – Just before radiation exposure • Cultured for 4 days • Quantified cell density using Crystal Violet Assay
    22. 22. Morphology Sham-EtOH Sham-VitD 2Gy-EtOH 2Gy-VitD
    23. 23. Results Pretreated EtOH Sham EtOH 2Gy VitD Sham VitD 2Gy 0 1 2 3 * * Treatment Absorbance590 Cotreated EtOH Sham EtOH 2Gy VitD Sham VitD 2Gy 0 1 2 3 * * Treatment Absorbance590
    24. 24. Conclusion • HME cells express VDR & p53 proteins • Radiation may increase VDR & p53 expression • VDR localization is altered by radiation treatment • Vitamin D inhibits growth of HME cells
    25. 25. Acknowledgments • Dr. JoEllen Welsh • Erika LaPorta • Dr. Meggan Keith • Dr. Judy Narvaez • Dr. Christopher Fernando • Deanna Morehead • UASRP Staff and Scholars

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