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)

Ichthyosis vulgaris is an epidermal disorder in which profilaggrin expression is decreased or absent. To determine whether the ichthyosis vulgaris phenotype could be mimicked by eliminating profilaggrin expression, a rat epidermal cell line was transfected with a plasmid that directs the constitutive expression of an RNA that is antisense to normal profilaggrin mRNA. Non-transfected and neomycin-resistant cells not containing antisense plasmid that were grown in the neomycin analogue G418 served as controls. Immunoblot and immunofluorescence analysis showed that profilaggrin protein expression and processing to filaggrin were delayed by 3 to 4 d and decreased in transfected cells. Profilaggrin mRNA was detected in both control and transfected cells only after the cells reached confluence, whereas antisense RNA was detected in transfectants at all times, even prior to confluence. Ultrastructural examination revealed that keratohyalin granules were decreased in number, globular, and heterogeneous in appearance in transfected cells in-contrast to angular structures seen in control cells. Unexpectedly, stratification was impaired, intermediate filaments were noticeably reduced, and cornified cell envelope formation was delayed in transfectants. Unlike ichthyosis vulgaris keratinocytes, where keratin expression is unaffected, appearance of K1 and K10 was delayed and K1/K10 synthesis was delayed and decreased in transfected cells. The precipitous drop in 35S-methionine incorporation into cytoskeletal protein seen at confluence in control cells was delayed by 3 d in transfected cells. We conclude that, rather than producing the ichthyosis vulgaris phenotype, antisense profilaggrin RNA has more broad-reaching effects on in vitro differentiation of rat epidermal keratinocytes.
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PMID:Antisense profilaggrin RNA delays and decreases profilaggrin expression and alters in vitro differentiation of rat epidermal keratinocytes. 768

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