Transformation of human urothelial cells (UROtsa) by as and cd induces the expression of keratin 6a.

Environmental Health Perspectives
Seema SomjiScott H Garrett

Abstract

Cadmium and arsenite can directly and malignantly transform the UROtsa cell line. The tumor heterotransplants produced from these transformed cells have histologic features consistent with human bladder cancer. Previous microarray analysis of total RNA from the parental and transformed cells suggested that keratin 6a was overexpressed as a result of cell transformation. Our goals were to verify overexpression of keratin 6a in Cd(2+)- and As(3+)-transformed UROtsa cells, the corresponding tumor heterotransplants, and human bladder cancer biopsy specimens and to assess what factors may be involved in keratin 6a overexpression. Expression was assessed with real-time polymerase chain reaction, Western blot analysis, and immunohistochemistry. We used the effect of addition and deletion of potential growth factors in the cell culture growth medium to assess possible pathways used in keratin 6a overexpression. Cd(2+)- and As(3+)-transformed cells grown in serum-containing growth medium, as well as the derived tumor heterotransplants, overexpressed keratin 6a mRNA and protein compared with UROtsa cells grown in serum-containing growth medium. Immunostaining of keratin 6a in tumor heterotransplants showed focal staining of the tumor cel...Continue Reading

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Methods Mentioned

BETA
chip
DNA shearing
protein assay
electrophoresis
light microscopy

Software Mentioned

Systat

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