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- Devangi Shah

"Combination of HDACi and Nucleoside Analog in inducing Apoptosis in Leukemia Cell Lines ". - Devangi Shah. I Thank. Dr. Steven Grant Dr. Roberto Rosato Jorge Almenara Stefanie Coe Dr. Allison Johnson, HHMI Coordinator. APOPTOSIS : Programmed cell death

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- Devangi Shah

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  1. "Combination of HDACi and Nucleoside Analog in inducing Apoptosis in Leukemia Cell Lines " - Devangi Shah

  2. I Thank • Dr. Steven Grant • Dr. Roberto Rosato • Jorge Almenara • Stefanie Coe • Dr. Allison Johnson, HHMI Coordinator

  3. APOPTOSIS : Programmed cell death HDAC : Histone deacetylases are a class of enzymes HDACI : HDAC Inhibitors are involved in anticancer activity LAQ824 : HDAC Inhibitor FLUDARABINE :A nucleoside analog U937 CELLS : Leukemia cell lines

  4. The Role of HDACs • A class of enzymes that remove acetyl groups from • lysine amino acid on a histone. • Altered expression and mutations of genes encoding • HDACs have been linked to tumor development

  5. The Role of LAQ824 • Inhibits cell growth in a dose- and time–dependent manner • Induces upregulation of p21, caspase cascade activation and Poly (ADP) ribose polymerase (PARP) cleavage

  6. The Role of Fludarabine • Inhibits DNA synthesis • Active against both dividing and resting cells. • One of the newer chemotherapy drugs which has been around and in use for a few years.

  7. My Project • Dose response of LAQ824 and Fludarabine to U937 cancer cells • Study of Proteins in U937 cells treated with LAQ824 and Fludarabine

  8. Dose Response of LAQ824 and Fludarabine to U937 cells

  9. Experiment • Treatments : • - Control • - LAQ824 (40 nM) • - FD 0.4, FD 0.5, FD 0.6, FD 0.7 (µM) • - L/F 0.4, L/F 0.5, L/F 0.6, L/F 0.7 • All LAQ824 treatments for 48 hours • All Fludarabine (FD) treatments for 24 hours

  10. Procedures Involved • Cells and Cell Culture • Drugs and Chemicals • Flow Cytometry

  11. Results

  12. Study of Proteins (p21, PARP, Caspase-3, Caspase-7 and Caspase-9)

  13. Experiment • Treatments • - Control • - LAQ824 40 nM • - FD 0.4 µM • - L/F 0.4

  14. Procedures Involved • Cells and Cell Culture • Drugs and Chemicals • Flow Cytometry • Western Blot Analysis

  15. Results Control LAQ824 FD 0.4 L/F 0.4 p21 PARP

  16. Caspase 9 CL40 4h 8h 16h 24h 28h 32h 40h 48h Cleavage Fragment C F0.4 4h 8h 16h 24h L/F 4h 8h 16h 24h

  17. Caspase 3 C L40 4h 8h 16h 24h 28h 32h 40h 48h C F0.4 4h 8h 16h 24h L/F 4h 8h 16h 24h

  18. Caspase 7 C L40 4h 8h 16h 24h 28h 32h 40h 48h Cleavage Fragment C F0.4 4h 8h 16h 24h L/F 4h 8h 16h 24h

  19. Conclusion LAQ824 and Fludarabine interact synergistically in U937 cancer cells inducing cell death by apoptotic events.

  20. Questions ??

  21. Thank You !!

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