Mechanisms of Disease

Follow

• 

  ITGB1 Knockdown Alters Apoptosis and Proliferation in Ewing Sarcoma Cells

  Aditi Kumar, Magdalena Saad, and Terry Jo Shackleford

  Integrin beta 1 (ITGB1) is an adhesion receptor that link cells to the extracellular matrix and regulates pathways involved in survival, proliferation, and migration. In order to understand its role in Ewing Sarcoma, we applied CRISPR-Cas9 with two guide RNAs to knock down ITGB1 in ES8 cells. We then assessed changes in cell growth, death, movement, and gene expression using Incucyte live-cell imaging, colony formation assays, wound healing assays, and qRT-PCR. qRT-PCR showed partial ITGB1 reduction, with ITGB1_gRNA2 producing the strongest decrease. ITGB1_gRNA2 also led to a small increase in caspase activity and a 20-30% increase in proliferation at the later time intervals. Migration remained unchanged, and colony formation varied with density but did not have statistical significance. Partial ITGB1 knockdown produced small but measurable effects on apoptosis and proliferation so ITGB1 does contribute to survival signaling in ES8 cells but may require extending imaging, improved qRT-PCR controls, and more replicates to help clarify these results.