Apoptosis in Proliferating, Senescent, and Immortalized Keratinocytes

• Published in: The Journal of biological chemistry Vol. 274; no. 33; pp. 23358 - 23367
• Main Authors: Chaturvedi, Vijaya, Qin, Jian-Zhong, Denning, Mitchell F., Choubey, Divaker, Diaz, Manuel O., Nickoloff, Brian J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 13.08.1999; American Society for Biochemistry and Molecular Biology
• Subjects: Apoptosis - radiation effects; Base Sequence; Cell Differentiation; Cell Division; Cell Line, Transformed; Cells, Cultured; Cellular Senescence; DNA Primers; Humans; Interferon-gamma - pharmacology; Keratinocytes - cytology; Keratinocytes - drug effects; Keratinocytes - radiation effects; NF-kappa B - antagonists & inhibitors; NF-kappa B - metabolism; Ultraviolet Rays
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.274.33.23358

Skin provides an attractive organ system for exploring coordinated regulation of keratinocyte (KC) proliferation, differentiation, senescence, and apoptosis. Our main objective was to determine whether various types of cell cycle arrest confer resistance to apoptosis. We postulated that KC cell cycle and cell death programs are tightly regulated to ensure epidermal homeostasis. In this report, simultaneous expression of cyclin-dependent kinase inhibitors (p15, p16, p21, and p27), a marker of early differentiation (keratin 1), mediators of apoptosis (caspases 3 and 8), and NF-κB were analyzed in three types of KCs. By comparing the response of proliferating, senescent, and immortalized KCs (HaCaT cells) to antiproliferative agents followed by UV exposure, we observed: 1) Normal KCs follow different pathways to abrupt cell cycle arrest; 2) KCs undergoing spontaneous replicative senescence or confluency predominantly express p16; 3) Abruptly induced growth arrest, confluency, and senescence pathways are associated with resistance to apoptosis; 4) The death-defying phenotype of KCs does not require early differentiation; 5) NF-κB is one regulator of resistance to apoptosis; and 6) HaCaT cells have undetectable p16 protein (hypermethylation of the promoter), dysfunctional NF-κB, and diminished capacity to respond to antiproliferative treatments, and they remain highly sensitive to apoptosis with cleavage of caspases 3 and 8. These data indicate that KCs (but not HaCaT cells) undergoing abruptly induced cell cycle arrest or senescence become resistant to apoptosis requiring properly regulated activation of NF-κB but not early differentiation.