Absolute quantification of senescence mediators in cells using multiple reaction monitoring liquid chromatography-Tandem mass spectrometry

Highlights: • Senescence role in multiple pathophysiological processes is controversial due to lacking distinctive biomarkers. • LC-MS/MS senescence profile that includes some current and proposed biomarkers was developed and validated. • Absolute quantification of senescence proteins in cancer and aging cell lines was performed. The identification of unique senescence markers remains challenging. Current hallmarks of senescent cells, including increased senescence-associated β-galactosidase activity, increased levels of cell cycle regulators such as p16^INK4a, p27, and p53, and altered levels of sirtuins and lamins, are detected commonly by Western blot and immunohistochemistry methods. Mass spectrometry outperforms these conventional quantification methods in terms of high throughput, specificity, and reproducibility.