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PTEN ablation in RasHa/Fos skin carcinogenesis invokes p53-dependent p21 to delay conversion while p53-independent p21 limits progression via cyclin D1/E2 inhibition

Oncogene volume 33, pages 4132–4143 (2014)Cite this article

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Abstract

To investigate tumour progression mechanism in transgenic mouse skin carcinogenesis, inducible PTEN ablation (Δ5PTENflx) was targeted to the epidermis of mice expressing activated rasHa/fos oncogenes (HK1.ras and HK1.fos). RU486-treated HK1.ras/fos-Δ5PTENflx epidermis exhibited significant keratinocyte proliferation resulting in hyperplasia and proliferating cysts. While HK1.ras/fos-Δ5PTENflx papillomatogenesis was accelerated, malignant conversion was delayed and tumours exhibited well-differentiated squamous cell carcinoma (wdSCC) histotypes, suggesting inhibition of early-stage malignant progression. Immediate elevated p53/p21 expression was observed in HK1.ras/fos-Δ5PTENflx hyperplasia, cysts and papillomas, and while malignant conversion required p53 loss, elevated p21 expression persisted in most wdSCCs to limit further progression, unless p21 was also lost and wdSCC progressed to more aggressive carcinomas. In contrast, TPA-promoted (that is, c-fos-activated) bi-genic HK1.ras-Δ5PTENflx cohorts lost p53/p21 expression during early papillomatogenesis and rapidly produced poorly differentiated carcinomas (pdSCCs) with high BrdU-labelling and elevated cyclin D1/E2 expression levels, indicative of a progression mechanism driven by failures in cell-cycle control. Intriguingly, HK1.ras/fos-Δ5PTENflx wdSCCs did not exhibit similar failures, as western and immunofluorescence analysis found downregulated cyclin E2 whenever p21 persisted; further, while westerns detected elevated cyclin D1, immunofluorescence identified reduced expression in proliferative basal layer nuclei and a redistributed expression profile throughout p21-positive wdSCC keratinocytes. These data demonstrate that rapid early epidermal responses to rasHa/fos/ΔPTEN co-operation involve induction of p53/p21 to alter differentiation and divert excessive proliferation into cyst formation. Further, despite three potent oncogenic insults p53 loss was required for malignant conversion, and following p53 loss persistent, p53-independent p21 expression possessed the potency to limit early-stage malignant progression via cyclin D1/E2 inhibition.

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Acknowledgements

We would like to thank Professor Hong Wu, Department of Molecular and Medical Pharmacology, UCLA, for the original gift of PTEN mice. Professor Dennis Roop, Center for Regenerative Medicine and Stem Cell Biology, Uiversity of Colorado, for the gift of K14.creP mice. Graham Chadwick for assistance with figure preparation; Dennis Duggan for assistance with animal husbandry and Ms Claire Gilbert for assistance with cyclin immunefluorescence analysis. This work was supported by: British Skin Foundation (Grants 610 & 3013), based on initial funding from Cancer Research UK (grant C1361/A2395) and the Scott Endowment to GU Dermatology.

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  1. F H Macdonald

    Present address: 3Current address: Department of Dermatology Monklands Hospital, Monkscourt Avenue, Airdrie ML6 0JS, UK.,

  2. D Yao

    Present address: 4Current address: Clinical Neurosciences, GBRC Building, Glasgow University, G12 8TA, UK.,

  3. F H Macdonald and D Yao: Co-first authors.

Authors and Affiliations

  1. Dermatology Section, School of Medicine, Dentistry and Nursing, College of Medical, Veterinary and Life Sciences, Glasgow University, Glasgow, UK

    F H Macdonald, D Yao, J A Quinn & D A Greenhalgh

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  1. F H Macdonald
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Correspondence to D A Greenhalgh.

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Macdonald, F., Yao, D., Quinn, J. et al. PTEN ablation in RasHa/Fos skin carcinogenesis invokes p53-dependent p21 to delay conversion while p53-independent p21 limits progression via cyclin D1/E2 inhibition. Oncogene 33, 4132–4143 (2014). https://doi.org/10.1038/onc.2013.372

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