Functional cooperation between ASK1 and p21Waf1/Cip1 in the balance of cell-cycle arrest, cell death and tumorigenesis of stressed keratinocytes

Both CDKN1A (p21 (Waf1/Cip1)) and Apoptosis signal-regulating kinase 1 (ASK1) play important roles in tumorigenesis. The role of p21 (Waf1/Cip1) in attenuating ASK1-induced apoptosis by various stress conditions is well established. However, how ASK1 and p21 (Waf1/Cip1) functionally interact during tumorigenesis is still unclear. To address this aspect, we crossed ASK1 knockout (ASK1KO) mice with p21(Waf1/Cip1) knockout (p21KO) mice to compare single and double-mutant mice. We observed that deletion of p21(Waf1/Cip1) leads to increased keratinocyte proliferation but also increased cell death. This is mechanistically linked to the ASK1 axis-induced apoptosis, including p38 and PARP. Indeed, deletion of ASK1 does not alter the proliferation but decreases the apoptosis of p21KO keratinocytes. To analyze as this interaction might affect skin carcinogenesis, we investigated the response of ASK1KO and p21KO mice to DMBA/TPA-induced tumorigenesis. Here we show that while endogenous ASK1 is dispensable for skin homeostasis, ASK1KO mice are resistant to DMBA/TPA-induced tumorigenesis. However, we found that epidermis lacking both p21 and ASK1 reacquires increased sensitivity to DMBA/TPA-induced tumorigenesis. We demonstrate that apoptosis and cell-cycle progression in p21KO keratinocytes are uncoupled in the absence of ASK1. These data support the model that a critical event ensuring the balance between cell death, cell-cycle arrest, and successful divisions in keratinocytes during stress conditions is the p21-dependent ASK1 inactivation.

Automated summary

The study investigates the functional interaction between ASK1 and p21 (Waf1/Cip1) in tumorigenesis. It shows that loss of p21 leads to increased keratinocyte proliferation and cell death, while loss of ASK1 reduces apoptosis in p21-deficient keratinocytes. Double knockout of p21 and ASK1 in skin epithelium increases sensitivity to carcinogens, indicating a critical role of p21-dependent ASK1 inactivation in maintaining a balance between cell death, cell-cycle arrest, and successful divisions during stress conditions.