Inorganic arsenic is a known human skin carcinogen. Chronic arsenic exposure results
in various human skin lesions, including hyperkeratosis and squamous cell carcinoma
(SCC), both characterized by distorted cytokeratin (CK) production. Prior work shows
the human skin keratinocyte HaCaT cell line, when exposed chronically for >25 weeks
to a low level of inorganic arsenite (100nM) results in cells able to produce aggressive
SCC upon inoculation into nude mice. In the present study, CK expression analysis
was performed in arsenic-exposed HaCaT cells during the progressive acquisition of
this malignant phenotype (0-20 weeks) to further validate this model as relevant to
epidermal carcinogenesis induced by arsenic in humans. Indeed, we observed clear evidence
of acquired cancer phenotype by 20 weeks of arsenite exposure including the formation
of giant cells, a >4-fold increase in colony formation in soft agar and a approximately
2.5-fold increase in matrix metalloproteinase-9 secretion, an enzyme often secreted
by cancer cells to help invade through the local extra-cellular matrix. During this
acquired malignant phenotype, various CK genes showed markedly altered expression
at the transcript and protein levels in a time-dependent manner. For example, CK1,
a marker of hyperkeratosis, increased up to 34-fold during arsenic-induced transformation,
while CK13, a marker for dermal cancer progression, increased up to 45-fold. The stem
cell marker, CK15, increased up to 7-fold, particularly during the later stages of
arsenic exposure, indicating a potential emergence of cancer stem-like cells with
arsenic-induced acquired malignant phenotype. The expression of involucrin and loricrin,
markers for keratinocyte differentiation, increased up to 9-fold. Thus, during arsenic-induced
acquired cancer phenotype in human keratinocytes, dramatic and dynamic alterations
in CK expression occur which are consistent with the process of epidermal carcinogenesis
helping validate this as an appropriate model for the study of arsenic-induced skin
cancer.