miR-425-5p Regulates Cellular Senescence Through Modulation of Retinoblastoma Protein Phosphorylation

MicroRNAs (miRNAs) are small non-coding RNAs that play critical roles in regulating cellular senescence and aging. Our recent studies identified a conserved C. elegans miRNA cluster (miR-229/64/65/66) that is required for normal adult lifespan, with overexpression significantly extending longevity. Notably, cel-miR-229 is evolutionarily conserved in humans, with hsa-miR-425 sharing an identical seed sequence.

Here, we investigated the role of miR-425 in mammalian cellular senescence. We found that miR-425 expression is markedly reduced in pharmacologically induced senescence in human lung cancer cells. Restoration of miR-425 expression attenuates senescence and suppresses the expression of senescence-associated secretory phenotype (SASP) cytokines following senescence induction. We further observed that miR-425 levels decline during replicative senescence, whereas stable overexpression in WI-38 fibroblasts delays senescence accumulation and preserves proliferative capacity.

Mechanistically, miR-425 suppresses TGF-β signaling, leading to reduced expression of the cyclin-dependent kinase inhibitor p21/ CDKN1A and increased phosphorylation of the retinoblastoma (RB) protein, thereby promoting cell-cycle progression. We further identify PPP2CB, the catalytic subunit of protein phosphatase 2A (PP2A), as a direct target of miR-425. PPP2CB expression is downregulated in miR-425-5p overexpressing cells, even under senescence induction. Knockdown of PPP2CB using siRNA phenocopies the effects of miR-425 overexpression, reducing senescence, enhancing proliferative potential, and increasing RB phosphorylation.

Collectively, our findings identify miR-425 as a conserved regulator of cellular senescence that acts through upregulation of RB phosphorylation. These results establish a novel miR-425- PPP2CB -RB regulatory axis controlling proliferation and senescence and suggest miR-425 as a potential therapeutic target for mitigating senescence to promote extended health span.