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Mol. Cell. Biol., 03 1995, 1175-1181, Vol 15, No. 3
J Kawanishi, J Kato, K Sasaki, S Fujii, N Watanabe and Y Niitsu
Detachment of cell-cell adhesion is indispensable for the first step of
invasion and metastasis of cancer. This mechanism is frequently associated
with the impairment of either E-cadherin expression or function. However,
mechanisms of such abnormalities have not been fully elucidated. In this
study, we demonstrated that the function of E- cadherin was completely
abolished in the human gastric cancer cell line HSC-39, despite the high
expression of E-cadherin, because of mutations in one of the
E-cadherin-associated cytoplasmic proteins, beta-catenin. Although
immunofluorescence staining of HSC-39 cells by using an anti-E- cadherin
antibody (HECD-1) revealed the strong and uniform expression of E-cadherin
on the cell surface, cell compaction and cell aggregation were not observed
in this cell. Western blotting (immunoblotting) using HECD-1 exhibited a
120-kDa band which is equivalent to normal E- cadherin. Northern (RNA)
blotting demonstrated a 4.7-kb band, the same as mature E-cadherin mRNA.
Immunoprecipitation of metabolically labeled proteins with HECD-1 revealed
three bands corresponding to E-cadherin, alpha-catenin, and gamma-catenin
and a 79-kDa band which was apparently smaller than that of normal
beta-catenin, indicating truncated beta- catenin. The 79-kDa band was
immunologically identified as beta-catenin by using immunoblotting with
anti-beta-catenin antibodies. Examination of beta-catenin mRNA by the
reverse transcriptase-PCR method revealed a transcript which was shorter
than that of normal beta-catenin. The sequencing of PCR product for
beta-catenin confirmed deletion in 321 bases from nucleotides +82 to
+402.(ABSTRACT TRUNCATED AT 250 WORDS)
Copyright © 1995, American Society for Microbiology
Loss of E-cadherin-dependent cell-cell adhesion due to mutation of the beta-catenin gene in a human cancer cell line, HSC-39
Fourth Department of Internal Medicine, Sapporo Medical University School of Medicine, Japan.
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