Photogenotoxicity and apoptosis in human HaCaT keratinocytes induced by 8-methoxypsoralen and lomefloxacin

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The HaCaT cell line is a spontaneously transformed human epithelial cell line from adult skin, which retain some differentiation characteristics of human skin cells. These cells are considered to more closely resemble the in vivo skin condition than other cell lines which are usually employed in genetic toxicity testing and may therefore be particularly appropriate for in vitro photogenotoxicity testing. We investigated chromosomal damage as evidenced by micronucleus (MN) formation in cytokinesis-blocked HaCaT keratinocytes after treatment with 8-methoxypsoralen (8-MOP) or lomefloxacin plus UV irradiation with a UVB/UVA relationship of 1:30. Our results show that 8-MOP induced MN in a dose-dependent manner over a concentration range from 1×10^<-7> to l×10^<-4> M using low and intermediate UVA/UVB irradiation dose levels. Lomefloxacin (LOM) induced MN in HaCaT cells in the range from 1×10^<-5> to 1×10^<-4> M only in conjunction with high UVA/UVB irradiation doses at which 8-MOP was too toxic to the cells. Both, photoactivated 8-MOP and LOM were also found to be strong inducers of apoptosis in HaCaT cells, probably as a consequence of their DNA damaging activities. Apoptosis was analysed at different days of cell culture, i.e., day 3, 7 or 11, at which HaCaT cells proceed from a proliferation state to differentiation. The highest sensitivity towards apoptosis was found in day-11 HaCaT cells suggesting that keratinocytes become increasingly prone to apoptosis during the differentiation process. In summary, our studies indicate that HaCaT cells are suitable target cells to assess multiple cellular effects, such as clastogenicity, cytotoxicity and apoptosis in response to UV-irradiated photosensitizers.
日本環境変異原学会の論文
2001-10-20

Photogenotoxicity and apoptosis in human HaCaT keratinocytes induced by 8-methoxypsoralen and lomefloxacin

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