Use of RT-PCR and DNA microarrays to characterize RNA recovered by non-invasive tape harvesting of normal and inflamed skin.
We describe a non-invasive approach for recovering RNA from the surface of skin via a simple tape stripping procedure that permits a direct quantitative and qualitative assessment of pathologic and physiologic biomarkers. Using semi-quantitative RT-PCR we show that tape-harvested RNA is comparable in quality and utility to RNA recovered by biopsy. It is likely that tape-harvested RNA is derived from epidermal cells residing close to the surface and includes adnexal structures and present data showing that tape and biopsy likely recover different cell populations. We report the successful amplification of tape-harvested RNA for hybridization to DNA microarrays. These experiments showed no significant gene expression level differences between replicate sites on a subject and minimal differences between a male and female subject. We also compared the array generated RNA profiles between normal and 24 h 1% SLS-occluded skin and observed that SLS treatment resulted in statistically significant changes in the expression levels of more than 1,700 genes. These data establish the utility of tape harvesting as a non-invasive method for capturing RNA from human skin and support the hypothesis that tape harvesting is an efficient method for sampling the epidermis and identifying select differentially regulated epidermal biomarkers.
Wong
Rita
R
DermTech International, Avenue of Science, San Diego, California 92128, USA.
Tran
Vynga
V
Morhenn
Vera
V
Hung
She-Pin
SP
Andersen
Bogi
B
Ito
Elaine
E
Wesley Hatfield
G
G
Benson
Nicholas R
NR
D000199:Actins / Q000235:genetics; D000328:Adult; D001706:Biopsy; D003872:Dermatitis / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology*; D005260:Female; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics; D016209:Interleukin-8 / Q000235:genetics; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000293:injuries / Q000503:physiopathology; D012879:Skin Physiological Phenomena / Q000235:genetics*
D000199:Actins; D007375:Interleukin-1; D016209:Interleukin-8; D012333:RNA, Messenger