Abstract
Human and canine mammary tumours show partial claudin expression deregulations. Further, claudins have been used for directed therapeutic approaches. However, the development of claudin targeting approaches requires stable claudin expressing cell lines. This study reports the establishment and characterisation of canine mammary tissue derived cell lines, analysing longitudinally the claudin-1, -3, -4 and -7 expressions in original tissue samples, primary cultures and developed cell lines. Primary cultures were derived from 17 canine mammary tissues: healthy, lobular hyperplasia, simple adenoma, complex adenoma, simple tubular carcinoma, complex carcinoma, carcinoma arising in a benign mixed tumour and benign mixed tissue. Cultivation was performed, if possible, until passage 30. Claudin mRNA and protein expressions were analysed by PCR, QuantiGene Plex Assay, immunocytochemistry and immunofluorescence. Further, cytokeratin expression was analysed immunocytochemically. Cultivation resulted in 11 established cell lines, eight showing epithelial character. In five of the early passages the claudin expressions decreased compared to the original tissues. In general, claudin expressions were diminished during cultivation. Three cell lines kept longitudinally claudin, as well as epithelial marker expressions, representing valuable tools for the development of claudin targeted anti-tumour therapies.
Highlights
The claudin (CLDN) protein family is a major structural and functional component of the tight junction complex in epithelial and endothelial cells [1,2,3]
CLDN proteins act as a receptor for the enterotoxin of Clostridium perfringens (Clostridium perfringens enterotoxin, CPE), a feature that enables a directed CLDN targeting for therapeutic approaches [16,17]
Further experimental therapeutic approaches targeting human cancer cells include the use of the enterotoxin of Clostridium perfringens (CPE) in order to kill cancer cells by directed CLDN binding
Summary
The claudin (CLDN) protein family is a major structural and functional component of the tight junction complex in epithelial and endothelial cells [1,2,3]. CLDN proteins act as a receptor for the enterotoxin of Clostridium perfringens (Clostridium perfringens enterotoxin, CPE), a feature that enables a directed CLDN targeting for therapeutic approaches [16,17] This feature was already used to successfully kill human CLDN-3 and -4 expressing cells in vivo and in vitro [7,16,17,18,19]. Directed recombinant mutation of the CPE sparing the cytotoxic domain leads to a protein that allows a directed reversible binding of CLDN proteins without killing the targeted cell [19,20,21] This enables drug delivery to CLDN expressing cells [22]. Cell lines can serve in comparative cancer genetics and translational medicine, as reported for canine lung tumour derived cell lines, showing genetic and biochemical similarities to human lung tumours and promising effects of a drug effective in human tumours on these cell lines [26]
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