New NCI-N87-derived @entity1 gastric epithelial line after XXXX telomerase catalytic subunit over-expression.

@entity1 @entity11178 @entity5644 @entity2355 @entity25856 @entity21826 @entity1711 @entity5962 @entity5091 @entity5824 @entity27334 @entity35540 @entity1534 @entity6175 @entity31843 @entity71850 @entity16090 @entity2215
AIM: To establish a cellular model correctly mimicking the gastric epithelium to overcome the limitation in the study of @entity1534 ( @entity1534 ) infection. METHODS: Aiming to overcome this limitation, clones of the heterogenic cancer-derived @entity1 cell line were isolated, by stably-transducing it with the @entity1 @entity2215 (hTERT) catalytic subunit gene. The clones were first characterized regarding their cell growth pattern and phenotype. For that we measured the clones' adherence properties, expression of cell-cell junctions' markers ( @entity11178 and @entity5962 ) and ability to generate a sustained transepithelial electrical resistance. The gastric properties of the clones, concerning expression of mucins, zymogens and glycan contents, were then evaluated by @entity6175 and @entity1711 staining, @entity5644 Schiff (PAS) and PAS/Alcian Blue-staining, immunocytochemistry and Western blot. In addition, we assessed the usefulness of the hTERT-expressing gastric cell line for @entity1534 research, by performing co-culture assays and measuring the IL-8 secretion, by ELISA, upon infection with two @entity1534 strains differing in virulence. RESULTS: Compared with the parental cell line, the most promising NCI-hTERT-derived clones (CL5 and @entity27334 ) were composed of cells with homogenous phenotype, presented higher relative telomerase activities, better adhesion properties, ability to be maintained in culture for longer periods after confluency, and were more efficient in PAS-reactive mucins secretion. Both clones were shown to produce high amounts of @entity2355 , @entity5091 and @entity31843 . NCI-hTERT-CL5 mucins were shown to be decorated with blood group @entity16090 (BG-H), Lewis-x ( @entity5824 ), @entity25856 and Le(a) and, in a less extent, with BG-A antigens, but the former two antigens were not detected in the NCI-hTERT- @entity27334 . None of the clones exhibited detectable levels of @entity21826 nor sialylated @entity5824 and Le(a) glycans. Entailing good gastric properties, both NCI-hTERT-clones were found to produce @entity35540 and @entity1 @entity71850 . The progenitor-like phenotype of NCI-hTERT- @entity27334 cells was highlighted by large nuclei and by the apical vesicular-like distribution of mucin 5AC and @entity35540 , supporting the accumulation of mucus-secreting and zymogens-chief mature cells functions. CONCLUSION: These traits, in addition to resistance to microaerobic conditions and good responsiveness to @entity1534 co-culture, in a strain virulence-dependent manner, make the NCI-hTERT- @entity27334 a promising model for future in vitro studies.
