Identification of cornifelin and early growth response-1 gene as novel biomarkers for in vitro eye irritation using a 3D reconstructed human cornea model MCTT HCE™
| Autor: | Su Hyon Lee, Young-Jin Chun, Seunghye Choi, Haeng Sun Jung, Kyung Min Lim, Miri Lee, Tae Young Chung, Seol Yeong Kim, Tae boo Choe |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Pathology
medicine.medical_specialty Microarray Octoxynol Health Toxicology and Mutagenesis medicine.medical_treatment Biology Toxicology Models Biological Sensitivity and Specificity Transcriptome Benzalkonium chloride Surface-Active Agents Cornea medicine Humans RNA Messenger Corneal transplantation Cells Cultured Early Growth Response Protein 1 Oligonucleotide Array Sequence Analysis Epithelium Corneal Membrane Proteins Sodium Dodecyl Sulfate General Medicine Epithelium In vitro Up-Regulation medicine.anatomical_structure Irritants Immunohistochemistry sense organs Benzalkonium Compounds Biomarkers medicine.drug |
| Zdroj: | Archives of toxicology. 89(9) |
| ISSN: | 1432-0738 |
| Popis: | Evaluation of the eye irritation is essential in the development of new cosmetic products. Draize rabbit eye irritation test has been widely used in which chemicals are directly applied to rabbit eye, and the symptoms and signs of eyes are scored. However, due to the invasive procedure, it causes substantial pain and discomfort to animals. Recently, we reported in vitro eye irritation test method using a 3D human corneal epithelial model (MCTT HCE™) which is reconstructed from remaining human tissues after a corneal transplantation. This model exhibited an excellent predictive capacity for 25 reference chemicals (sensitivity 100 %, specificity 77 % and accuracy 88 % vs. GHS). To improve the test performance, we explored new biomarkers for the eye irritation through transcriptomic approach. Three surfactants were selected as model eye irritants that include sodium lauryl sulfate, benzalkonium chloride and triton X-100. After test chemicals were treated, we investigated differentially expressed genes through a whole-gene microarray (Affymetrix GeneChip® Human Gene 2.0 ST Array, 48,000 probes). As a result, we identified that mRNAs of cornifelin (CNFN), a constituent of the insoluble cornified cell envelope of stratified squamous epithelia, and early growth response-1 (EGR1), a nuclear transcriptional regulator, were significantly up-regulated by all three irritants. Up-regulation of CNFN and EGR1 was further confirmed by Q-RT-PCR, and immunohistochemistry revealed increased level of CNFN in irritant-treated tissues, supporting the relevance of CNFN and EGR1 as new biomarkers for eye irritation. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink