2014 Sigma Xi PowerPoint Presentation

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MUC1 as a biomarker for TGF- β1 inhibition: Investigating the role of MUC1 in the switch of TGF-β1 function from a tumor suppressor to a tumor promoter in pancreatic ductal adenocarcinoma. Emily Ashkin

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This research was carried out under Dr. Pinku Mukherjee and Dr. Sritama Nath at the University of North Carolina at Charlotte Biology Department.

Transcript of 2014 Sigma Xi PowerPoint Presentation

  • 1. MUC1 as a biomarker for TGF1 inhibition: Investigating the role of MUC1 in the switch of TGF-1 function from a tumor suppressor to a tumor promoter in pancreatic ductal adenocarcinoma. Emily Ashkin
  • 2. Pancreatic cancer is the most lethal of all cancers. 6% five-year-survival rate. Median survival of 6 months. 4th leading cause of cancer related deaths in the United States.
  • 3. Background: Pancreatic Ductal Adenocarcinoma Accounts for more than 95% of pancreatic cancer cases. Therapeutic interventions include surgical resection, radiation therapy, chemotherapy and immunotherapy Lack of specific symptoms and limitations in diagnostic methods enable cancer progression.
  • 4. Background: TGF-1 Original member of the Transforming Growth Factor Beta super family, consisting of three isoforms. Type of cytokine and secreted protein.
  • 5. Background: Dual Function of TGF-1 Tumor suppressor TGF-1 induces apoptosis, or cell suicide, by signaling the SMAD pathway As the tumor progresses, genetic and/or biochemical changes allow TGF-1 to stimulate tumor progression by its pleiotropic activities on the cancer cells. TGF-1, binding to receptor II, induces epithelial-tomesenchymal transition (EMT). TGF-1-induced EMT leads to migration and invasion of local epithelial cells
  • 6. Hypothesis Signaling through MUC1 inhibits TGF-1-induced-apoptosis and supports TGF-1-induced EMT and invasion.
  • 7. MUC1 A transmembrane mucin glycoprotein Over-expressed in more than 90% of pancreatic cancer Plays a major role in EMT as well as drug resistance, invasion, and metastasis in pancreatic cancer
  • 8. Methods and Materials Experiments Descriptions Cell Culture All cell lines were previously established cell lines ordered from the American Type Culture Collection (ATCC). Migration Scratch Assay The scratch assay is a simple, reproducible assay commonly used to measure basic cell migration parameters. This assay was performed in order to visibly see each cell's migratory capabilities and draw conclusions on metastasis. BCA Assay Biochemical assay for determining the amount of protein present in a solution. Western Blot Molecular weights: MUC1 (25 kDa), TGF-R1 (56 kDa), TGF-R2 (64 kDA), and -actin (42 kDa) TGF-1 Binding MUC1+ and MUC1- cells were treated with pure recombinant human TGF-1. Flow Cytometry Cell cycle analysis was performed using Propidium Iodide DNA staining.
  • 9. Images
  • 10. Pancreatic Cancer Human and Mouse Cell Lines
  • 11. Results: Migration Scratch Assay
  • 12. Results: Migration Scratch Assay
  • 13. Results: Migration Scratch Assay The most significant change in wound size is between 0 and 14 hours For both mouse and human cell lines, the MUC1+ cells were more migratory than the MUC1- cell lines Using Image J programming, I was able to measure the width of the wound and analyze the decrease in size or lack thereof.
  • 14. Results: Western Blot
  • 15. Results: Western Blot p