| 研究実績の概要 |
The purpose of this project is to identify the variant of CD44 recognized by KM48 antibody and to assess its functional role in normal and human skin cancer by various methods. Expression of more than 800 transcripts of CD44 is possible but not all are expressed and the expression is tissue and context dependent. Previously, we found 18 CD44 transcripts and believed one of them is related to KM48 antigen, which is lost in human SCC. We studied how expression of CD44 expressions are regulated and found that expression of certain CD44 molecules are regulated differentially between normal and malignant keratinocytes. Furthermore, specific growth promoting, REDOX-related, and differentiation-related molecules were found to differentially affect CD44 transcription. Thus, we were able to identify candidate CD44 transcripts that may be associated with KM48 antigen. Than, we transfected all the CD44 clones into optimized cells using optimized vectors and and optimized transfection reagent. Western blotting of transfected HEK293T cells successfully revealed the CD44 variant recognized by KM48 antibody. Coincidentally, the CD44 variant recognized by KM48 antibody is reduced in SCC cell lines at the mRNA levels. Thus, although the epitope for KM48 antibody is difficult to produce in cultured cells, by using optimized cell culture and expression systems, were able to identify the CD44 variant recognized by KM48 antibody. The results should aid further studies about the role of CD44 in maintaining normal skin barrier function to prevent cancer.
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