Absolute beta-catenin concentrations in Wnt pathway-stimulated and non-stimulated cells.
The intracellular level of the proto-oncoprotein beta-catenin is a parameter for the activity of the Wnt pathway, which has been linked to carcinogenesis. The paper introduces a novel sandwich-based ELISA for the determination of the beta-catenin concentration in lysates from cells or tissues. The advantages of the method were proven by determining beta-catenin levels in cell lines and in cells after activation of the Wnt pathway. Analysis revealed high beta-catenin concentrations in the cell lines HeLa, KB, HT1080, MCF-7, U-87 and U-373, which had not been described before. Beta-catenin concentrations were compared in HEK293 and C57MG cells after activation of the Wnt pathway. The beta-catenin concentrations increased by different factors depending on whether the Wnt pathway was activated by incubation with LiCl or with Wnt-3a-conditioned medium. This finding indicated that the beta-catenin level depends on the way and level of Wnt pathway activation. The quantitative analysis of beta-catenin in colorectal tumours revealed high beta-catenin levels in tumours with truncating mutations in the APC gene.
Sievers
S
S
Max-Planck-Institut für Molekulare Physiologie, Dortmund, Germany.
Fritzsch
C
C
Grzegorczyk
M
M
Kuhnen
C
C
Müller
O
O
D002460:Cell Line; D045744:Cell Line, Tumor; D002474:Cell-Free System; D015179:Colorectal Neoplasms / Q000175:diagnosis / Q000235:genetics; D017077:Culture Media, Conditioned; D004797:Enzyme-Linked Immunosorbent Assay; D017491:Genes, APC; D006801:Humans; D018021:Lithium Chloride; D009369:Neoplasms / Q000175:diagnosis; D051153:Wnt Proteins / Q000378:metabolism*; D051176:beta Catenin / Q000032:analysis*
D017077:Culture Media, Conditioned; D051153:Wnt Proteins; D051176:beta Catenin; D018021:Lithium Chloride