Replicative senescence in human uroepithelial cells

Abstract
PURPOSE:
Normal human uroepithelial cells (HUCs) proliferate rapidly in culture during early passage and then spontaneously undergo replicative senescence. We previously reported that the cyclin D1-CDK4/6 inhibitor, p16INK4a, is elevated at senescence in HUCs. Hence, we proposed that p16INK4a may play a critical role in mediating senescence in this cell type. In the current study, we further characterized the senescent state in HUCs. We also tested the possible roles of changes in other cell cycle proteins, including p53, p21WAF1, pRb, and cyclin D1 in HUC senescence.

These cells are the ones lining the urinary bladder and have high plasticity and perform a variety of cell functions. They perform bladder defense and act as an interface between pathogens. The receptors which these cells inhibit are very crucial to analyze the other factors related to these cells’ reaction to injury or infection. They are also responsible for factors that keep the immune system intact. The further study and analysis of these cells help in developing the immune system regulatory levels and pose a necessity in the current world. Immunity is very important to a human being as it is the capability to automatically prevent any disease or infection, this can be vaguely compared to the modern day online trading software like ethereum code which is autoimmune to huge losses. Thus the study of these cells have a long way to go and simple understanding of the same is given below.

METHODS:
Normal HUCs cultured from explants of ureteral mucosa were used for these studies. Senescence associated-beta-galactosidase activity (SA-beta-gal) was used to identify cells in senescence. Flow cytometric analysis was used to determine changes in cell cycle distribution at senescence. Response of cells to serum stimulation was determined by Northern analysis of c-fos. Western analysis was used to assess changes in p53, p21WAF, p16INK4a, cyclin D1 and plasminogen activator inhibitor-1 (PAI-1) levels at senescence.

RESULTS:
beta-gal-positive HUCs were blocked at G1/S in senescence and failed to show c-fos induction in response to serum stimulation. As previously reported, senescent HUCs also showed elevated p16INK4a. However, unlike human fibroblasts, neither p53 nor p21WAF1 elevation accompanied HUCs senescence. PAI-1 levels were also not elevated in HUC senescence.

CONCLUSION:
These findings support a model in which elevation of p16INK4a, but not p53 or p21WAF1 plays a critical role in HUC replicative senescence. These findings elucidate the tumor suppressor mechanism of p16INK4a and the frequent loss of either p16INK4a or pRb in invasive human bladder tumors.