Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT01586 (G418)
 2,237 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have isolated rabbit kidney proximal tubular epithelial cell lines. The selection was based on their ability to form confluent monolayers on porous supports and to maintain receptor-mediated signal transduction and ion transport, characteristic of the proximal tubule. The isolation method consisted of several steps: (1) superficial cortical proximal tubule segments were microdissected and cultured on a matrix-coated porous support until cells formed a confluent monolayer; (2) primary cultures showing hormone-regulated ion transport typical for the proximal tubule were selected and co-cultured with irradiated fibroblasts; and (3) the epithelial cells surviving after several passages were expanded and passaged on porous substrates. Most of the cell lines developed in this manner were obtained by co-culture with irradiated fibroblasts producing a recombinant retrovirus encoding SV40 large T antigen and G418 resistance. However, SV40 T antigen expression was not essential for immortalization, since neither T antigen nor G418 resistance was detected in the isolated cell lines and co-culture with non-producing 3T3 cells gave similar results. One cell line (vEPT) has been characterized in some detail with respect to morphological, biochemical, and ion transport properties. This line forms confluent monolayers with apical microvilli, tight junctions, and convolutions of the basolateral plasma membrane. Once confluent, monolayers maintain conductances of 25 to 32 mS/cm2 for several weeks in culture and possess phlorizin-sensitive short-circuit current (Isc) in glucose containing media, indicative of apical Na(+)-glucose co-transport. vEPT cells also retain receptor and signaling mechanisms for angiotensin II (Ang II). Apical and basal Ang II and 5,6-epoxy-eicosatrienoic acid (5,6-EET) modulate the Isc in a manner similar to primary cultures. The cell lines share with primary cultures expression of the cytokeratins K8, K10/K11, and K19 ("nomenclature" [21]). They also retain several receptor and signal transduction mechanisms. For example, Ang II, arachidonate, bradykinin, 5,6-EET, parathyroid hormone (residues 1 through 34), and purine nucleotides increase cytosolic Ca2+, PTH elevates cAMP levels, and Ang II enhances proximal tubule-specific arachidonic acid metabolism.
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PMID:Development and characterization of rabbit proximal tubular epithelial cell lines. 128 Jul 3

Human papillomavirus (HPV) has been implicated in the etiology of oral and cervical cancers. Normal oral epithelial cells at passage two were infected with recombinant retrovirus containing the E6/E7 open reading frames of HPV type 16. The G418-selected cells that were immortalized and express HPV 16 E6/E7 have been in culture for over 4 years and 350 passages. In contrast, the normal oral epithelial cells did not survive the culture environment beyond 7 to 9 passages. Fifteen clones were selected from the pooled population. By Northern blot analysis all clones demonstrated the presence of E6solidusE7 genes. Keratin expression of both normal and immortalized oral epithelial cells was studied in organotypic culture. Both cell types were positive with antibodies AE1, AE3, and 34BE12. Both were focally positive with AE8, which stains for keratin 13 (specific for oral and esophageal epithelial cells). The normal control cells were focally positive for filaggrin, while the HPV 16-immortalized cells (IHGK cells-immortalized human gingival keratinocytes) were negative. The IHGK cells were strongly positive with KS19.1, which stains the embryonal keratin K19, an indicator of premalignant or malignant changes, while the normal control cells were only lightly and focally positive. In conclusion, we present an oral epithelial cell line successfully immortalized with HPV E6/E7 which will facilitate further research on the involvement of HPV in oral carcinogenesis.
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PMID:HPV immortalization of human oral epithelial cells: a model for carcinogenesis. 866 Sep 52