Ectopic expression of gastrokine 1 in gastric cancer cells up-regulates tight and adherens junction proteins network

Emilia Rippa, Filomena Altieri, Chiara Stella Di Stadio, Giuseppina Miselli, Annalisa Lamberti, Antonella Federico, Vincenzo Quagliariello, Ferdinando Papale, Germano Guerra, Paolo Arcari

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Gastrokine 1 (GKN1) is a stomach-specific protein important in the replenishment of the surface lumen epithelial cell layer and in maintaining mucosal integrity. A role in cell proliferation and differentiation has also been hypothesized. Despite these findings, the function(s) as well as the cellular localization of GKN1 in the cellular machinery are currently not clarified. The investigation of subcellular localization of GKN1 in gastric cancer cells can provide insights into its potential cellular roles. Subcellular fractions of gastric cancer cells (AGS) transfected with full-length GKN1 (flGKN1) or incubated with recombinant GKN1 (rGKN1) lacking the first 20 amino acids at N-terminal were analyzed by Western blot and confocal microscopy and compared with those from normal gastric tissue. Wild type GKN1 (wtGKN1) and flGKN1 were revealed in the cytoplasm and in the membrane fractions of gastric cells, whereas rGKN1 was revealed in the cytoplasmic fractions, but a high amount was detected in the membrane pellet of the AGS lysate. The cellular distribution of GKN1 was also confirmed by confocal microscopy. The purified protein was also used to highlight its possible association with actin through confocal microscopy, pelleting assay, and size-exclusion chromatography. GKN1 co-localizes with actin in normal gastric tissue, but no direct interaction was observed between the two proteins in vitro. Most likely, GKN1 indirectly participates in actin stabilization since its overexpression in gastric cancer cells strongly increases the expression of tight and adherens junction proteins.

Original languageEnglish
Pages (from-to)577-583
Number of pages7
JournalPathology Research and Practice
Volume211
Issue number8
DOIs
Publication statusPublished - Aug 1 2015

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Tight Junction Proteins
Adherens Junctions
Stomach Neoplasms
Stomach
Up-Regulation
Confocal Microscopy
Actins
Proteins
Subcellular Fractions
Membranes
Gel Chromatography
Cell Differentiation
Cytoplasm
Western Blotting
Epithelial Cells
Cell Proliferation
Amino Acids
Ectopic Gene Expression

Keywords

  • Actin
  • Cellular localization
  • Confocal microscopy
  • Gastric cancer
  • Gastrokine 1

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology

Cite this

Rippa, E., Altieri, F., Di Stadio, C. S., Miselli, G., Lamberti, A., Federico, A., ... Arcari, P. (2015). Ectopic expression of gastrokine 1 in gastric cancer cells up-regulates tight and adherens junction proteins network. Pathology Research and Practice, 211(8), 577-583. https://doi.org/10.1016/j.prp.2015.04.008

Ectopic expression of gastrokine 1 in gastric cancer cells up-regulates tight and adherens junction proteins network. / Rippa, Emilia; Altieri, Filomena; Di Stadio, Chiara Stella; Miselli, Giuseppina; Lamberti, Annalisa; Federico, Antonella; Quagliariello, Vincenzo; Papale, Ferdinando; Guerra, Germano; Arcari, Paolo.

In: Pathology Research and Practice, Vol. 211, No. 8, 01.08.2015, p. 577-583.

Research output: Contribution to journalArticle

Rippa, E, Altieri, F, Di Stadio, CS, Miselli, G, Lamberti, A, Federico, A, Quagliariello, V, Papale, F, Guerra, G & Arcari, P 2015, 'Ectopic expression of gastrokine 1 in gastric cancer cells up-regulates tight and adherens junction proteins network', Pathology Research and Practice, vol. 211, no. 8, pp. 577-583. https://doi.org/10.1016/j.prp.2015.04.008
Rippa, Emilia ; Altieri, Filomena ; Di Stadio, Chiara Stella ; Miselli, Giuseppina ; Lamberti, Annalisa ; Federico, Antonella ; Quagliariello, Vincenzo ; Papale, Ferdinando ; Guerra, Germano ; Arcari, Paolo. / Ectopic expression of gastrokine 1 in gastric cancer cells up-regulates tight and adherens junction proteins network. In: Pathology Research and Practice. 2015 ; Vol. 211, No. 8. pp. 577-583.
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