Two-dimensional gel electrophoretic studies on the cellular aging: Accumulation of alpha-2-macroglobulin in human fibroblasts with aging

To understand the mechanisms that control life span and age-related phenotypes, we used two-dimensional (2D) gel electrophoresis to study the intracellular proteins whose amounts change during the process of cellular aging. We found that the amount of an alpha-2-macroglobulin (A2M) fragment derived from culture medium increased in the cells with aging. A2M is linked to Alzheimer's disease both genetically and functionally. This is the first report of accumulation of an A2M fragment in senescent fibroblasts. We also studied 2D gel profiles of human fibroblasts immortalized by treatment with either ⁶⁰Co gamma rays or 4-nitroquinoline 1-oxide. As immortalized cells overcome cellular senescence to gain an unlimited life span, the proteins whose amounts change after immortalization may be relevant to the age-related phenotypes. 2D gel analysis revealed that the A2M fragment was down-regulated in the immortalized cells, compared with their normal counterparts, regardless of their passage. We also found that the other four proteins increased in amount with aging and decreased in amount after immortalization. Our results suggest: (1) the A2M incorporation into the cells is increased in the process of cellular aging; and (2) A2M may be linked to the age-related phenotypes that were lost during the process of immortalization of human cells.