Journal Article

Apoptosis and cytokine release induced by ionizing or ultraviolet B radiation in primary and immortalized human keratinocytes

Corinne Petit-Frère, Emily Capulas, Debbie A. Lyon, Christopher J. Norbury, Jillian E. Lowe, Peter H. Clingen, Enriqueta Riballo, Michael H. L. Green and Colin F. Arlett

in Carcinogenesis

Volume 21, issue 6, pages 1087-1095
Published in print June 2000 | ISSN: 0143-3334
Published online June 2000 | e-ISSN: 1460-2180 | DOI:
Apoptosis and cytokine release induced by ionizing or ultraviolet B radiation in primary and immortalized human keratinocytes

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We have compared the induction of apoptosis and cytokine release by UVB and γ-radiation in primary (untransformed) and in two immortalized human epithelial/keratinocyte cell lines, HaCaT and KB (KB is now known to be a subline of the ubiquitous keratin-forming tumour cell line HeLa and we therefore designate it HeLa-KB). In both the primary and the immortalized cell lines apoptosis and release of the inflammatory cytokine interleukin-6 are induced rapidly following UVB irradiation. In contrast, only the immortalized cells undergo apoptosis and release interleukin-6 after γ-irradiation and here the onset of apoptosis and cytokine release are delayed. The same distinction between primary and immortalized cells was observed when double-strand breaks were induced with the anticancer drug mitoxantrone, which stabilizes topoisomerase II-cleavable complexes. We suggest that immortalization may sensitize keratinocytes to the apoptogenic effect of ionizing radiation or mitoxantrone by deregulating normal cell cycle checkpoints. In both human keratinocytes and fibroblasts, cell killing, as assayed by loss of colony-forming ability, is not coupled to apoptosis. Immortalization increases resistance to γ-radiation killing but sensitizes to apoptosis. In contrast, although immortalization also sensitizes to UVB-induced apoptosis, it does not affect UVB-induced cell killing. Apoptosis unambiguously indicates death at the single cell level but clonal cell survival integrates all the cellular and genetic processes which prevent or permit a scorable clone to develop.

Keywords: DSB, double-strand break; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IL-6, interleukin-6; NHEK, normal human epidermal keratinocytes; RT–PCR, reverse transcription–PCR; TGFβ, transforming growth factor-β; TNFα, tumour necrosis factor α; VDM, time lapse video microscopy.

Journal Article.  6612 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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