Due to their limited replicative lifespan, normal cells eventually enter a state of cell cycle arrest known as cellular senescence. During senescence, cells remain metabolically active but do not undergo cell death or division. These senescent cells exhibit a distinct phenotype, characterized by the appearance of multinucleated cells, increased vacuolization, expression of pH-dependent β-galactosidase, and morphological changes such as enlargement and elongation. Senescence can influence various biological processes, including tumor suppression, tumor progression, aging, and tissue repair.
β-galactosidase (β-gal) is a crucial hydrolase enzyme that catalyzes the hydrolysis of galactose-containing carbohydrates into monosaccharides. Its substrates include lactose, various glycoproteins, ganglioside GM1, and lactosylceramides. In molecular biology, β-galactosidase is widely used; for example, the isolation of recombinant bacteria during molecular cloning involves α-complementation of the bacterial β-galactosidase gene (lacZ) in the presence of a β-gal substrate to identify recombinant clones. In cell biology, Senescence-Associated beta-galactosidase (SA-β-gal), defined as β-gal activity at pH 6.0, is a widely used marker of replicative senescence.
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