The detachment and subsequent death of EO771 cells following gene knockout or knockdown likely point to loss of adhesion-dependent survival signaling, a process known as anoikis. This occurs when cells lose contact with the extracellular matrix (ECM), especially due to disruption of integrin-mediated signaling pathways. If the targeted gene is involved in focal adhesion, ECM interaction (e.g., integrins, FAK, SRC), or cytoskeletal dynamics, its suppression could trigger this phenotype. The recurrence of cell death after reaching high confluency or after passaging supports a mechanism involving stress-induced detachment or disrupted cell-matrix interactions.

I recommend:

• Assessing expression of integrin subunits (e.g., ITGB1), focal adhesion kinase (FAK), and E-cadherin.
• Performing rescue experiments or culturing on ECM-coated plates (like fibronectin or collagen).
• Testing for cleaved caspase-3 or annexin V/PI staining to confirm apoptosis.

Reference: Paoli, P., Giannoni, E., & Chiarugi, P. (2013). Anoikis molecular pathways and its role in cancer progression. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1833(12), 3481–3498. https://doi.org/10.1016/j.bbamcr.2013.06.026