Cytoskeleton reorganization and ultrastructural damage induced by gliadin in a three-dimensional in vitro model.
OBJECTIVE
To evaluate the interplay between gliadin and LoVo cells and the direct effect of gliadin on cytoskeletal patterns.


METHODS
We treated LoVo multicellular spheroids with digested bread wheat gliadin in order to investigate their morphology and ultrastructure (by means of light microscopy and scanning electron microscopy), and the effect of gliadin on actin (phalloidin fluorescence) and the tight-junction protein occludin and zonula occluden-1.


RESULTS
The treated spheroids had deep holes and surface blebs, whereas the controls were smoothly surfaced ovoids. The incubation of LoVo spheroids with gliadin decreased the number of intracellular actin filaments, impaired and disassembled the integrity of the tight-junction system.


CONCLUSIONS
Our data obtained from an "in vivo-like" polarized culture system confirm the direct noxious effect of gliadin on the cytoskeleton and tight junctions of epithelial cells. Unlike two-dimensional cell culture systems, the use of multicellular spheroids seems to provide a suitable model for studying cell-cell interactions.
Dolfini
Ersilia
E
Department of Biology and Genetics for Health Sciences, Via Viotti 3/5, 20131 Milano, Italy. ersilia.dolfini@unimi.it
Roncoroni
Leda
L
Elli
Luca
L
Fumagalli
Chiara
C
Colombo
Roberto
R
Ramponi
Simona
S
Forlani
Fabio
F
Bardella
Maria Teresa
MT
D002450:Cell Communication / Q000187:drug effects; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000187:drug effects* / Q000648:ultrastructure; D005903:Gliadin / Q000633:toxicity*; D006801:Humans; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D018874:Spheroids, Cellular; D019108:Tight Junctions / Q000187:drug effects / Q000648:ultrastructure
D005903:Gliadin