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TWIST and Senescence

TWIST and Senescence. Emily McMillan 1/22/08. Differentiation. Transcription factor bLHL Proliferation factor Binds RUNX2/CBFA1 Inhibits osteoblast differentiation. Rose and Malcom. 1997 TIG: 384. Aging. TWIST differentially expressed Young fibroblasts = ↑ TWIST1

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TWIST and Senescence

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  1. TWIST and Senescence Emily McMillan 1/22/08

  2. Differentiation • Transcription factor • bLHL • Proliferation factor • Binds RUNX2/CBFA1 • Inhibits osteoblast differentiation Rose and Malcom. 1997 TIG: 384.

  3. Aging • TWIST differentially expressed • Young fibroblasts = ↑ TWIST1 • Senescent fibroblasts = ↓ TWIST1

  4. Do senescent fibroblasts have an osteoblast-like phenotype?

  5. Cell Types • WI-38 (lung fibroblasts) • AG13153 (skin fibroblasts) • NHOst • U2Os (osteosarcoma) • BMSC (+/- differentiation factors)

  6. Gene Markers • Twist • Beta-2-microglobin (normalization) • Osteogenic Markers • Runx2 • Osterix • Osteocalcin • Osteopontin • Osteonectin • Bone Sialoprotein • Alkaline Phospatase • Collagen Type 1A

  7. Analyses • Real-Time PCR • Immunofluorescence • Western blot

  8. Northern vs. Real-Time Wang et al (1996)

  9. RUNX2 Levels

  10. Collagen 1A Levels

  11. Osteonectin Levels

  12. Osteopontin Levels

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