Variable expression of some "housekeeping" genes during human keratinocyte differentiation

被引:70
作者
Steele, BK
Meyers, C
Ozbun, MA
机构
[1] Univ New Mexico, Sch Med, Dept Mol Genet & Microbiol, Albuquerque, NM 87131 USA
[2] Penn State Univ, Milton S Hershey Med Ctr, Coll Med, Dept Microbiol & Immunol, Hershey, PA 17033 USA
关键词
epithelial differentiation; GAPDH; 28S rRNA; cyclophilin; beta-actin; human papillomavirus;
D O I
10.1016/S0003-2697(02)00045-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We investigated the expression levels of four cellular "housekeeping" genes during epithelial differentiation. Differentiation is a dynamic process and various cellular RNAs have been targeted for use as internal controls during differentiation of human keratinocytes, but the consistent expression of such standards has not been previously validated. We used the organotypic (raft) culture system to grow stratified and differentiated epithelium in vitro. We compared cellular RNAs from epithelial tissues of both normal human keratinocytes and keratinocytes whose differentiation scheme is altered by the replication of human papillomavirus. Using ribonuclease protection assays to quantify RNA expression levels, we found that beta-actin and glyceraldehyde-3-phosphate dehydrogenase levels fluctuated during epithelial differentiation, whereas cyclophilin RNA and 28S-ribosomal RNA were the most consistently expressed during epithelial differentiation. These stably expressed cellular RNAs can be targeted as controls to permit quantitative expression analyses of cellular and pathogen RNAs during epithelial differentiation under various experimental conditions. (C) 2002 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:341 / 347
页数:7
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