title: Functional protein pathway activation mapping of the progression of normal skin to squamous cell carcinoma.
abstract: Reverse phase protein microarray analysis was used to identify cell signaling derangements in squamous cell carcinoma (SCC) compared with actinic keratosis (AK) and upper inner arm (UIA). We analyzed two independent tissue sets with isolation and enrichment of epithelial cells by laser capture microdissection. Set 1 served as a pilot and a means to identify protein pathway activation alterations that could be further validated in a second independent set. Set 1 was comprised of 4 AK, 13 SCC, and 20 UIA. Set 2 included 15 AK, 9 SCCs, and 20 UIAs. Activation of 51 signaling proteins, known to be involved in tumorigenesis, were assessed for set 1 and showed that the MEK-ERK [mitogen-activated protein (MAP)/extracellular signal-regulated (ERK; MEK)] pathway was activated in SCC compared with AK and UIA, and that epidermal growth factor receptor (EGFR) and mTOR pathways were aberrantly activated in SCC. Unsupervised two-way hierarchical clustering revealed that AK and UIA shared a common signaling network activation architecture while SCC was dramatically different. Statistical analysis found that prosurvival signaling through phosphorylation of ASK and 4EBP1 as well as increased Bax and Bak expression was higher in AK compared with UIA. We expanded pathway network activation mapping in set 2 to 101 key signaling proteins, which corroborated activation of MEK-ERK, EGFR, and mTOR pathways through discovery of a number of upstream and downstream signaling molecules within these pathways to conclude that SCC is indeed a pathway activation-driven disease. Pathway activation mapping of SCC compared with AK revealed several interconnected networks that could be targeted with drug therapy for potential chemoprevention and therapeutic applications.
authors.affiliation: University of Arizona Cancer Center, 1515 North Campbell Avenue, Tucson, AZ 85724, USA. jeinspahr@azcc.arizona.edu
authors.forename: Janine G
authors.initials: JG
authors.lastname: Einspahr
authors.forename: Valerie
authors.initials: V
authors.lastname: Calvert
authors.forename: David S
authors.initials: DS
authors.lastname: Alberts
authors.forename: Clara
authors.initials: C
authors.lastname: Curiel-Lewandrowski
authors.forename: James
authors.initials: J
authors.lastname: Warneke
authors.forename: Robert
authors.initials: R
authors.lastname: Krouse
authors.forename: Steven P
authors.initials: SP
authors.lastname: Stratton
authors.forename: Lance
authors.initials: L
authors.lastname: Liotta
authors.forename: Caterina
authors.initials: C
authors.lastname: Longo
authors.forename: Giovanni
authors.initials: G
authors.lastname: Pellacani
authors.forename: Giovanni
authors.initials: G
authors.lastname: Pellicani
authors.forename: Anil
authors.initials: A
authors.lastname: Prasad
authors.forename: Paul
authors.initials: P
authors.lastname: Sagerman
authors.forename: Yira
authors.initials: Y
authors.lastname: Bermudez
authors.forename: Jianghong
authors.initials: J
authors.lastname: Deng
authors.forename: G Timothy
authors.initials: GT
authors.lastname: Bowden
authors.forename: Emanuel F
authors.initials: EF
authors.lastname: Petricoin
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D018450:Disease Progression; D005260:Female; D006801:Humans; D061346:Laser Capture Microdissection; D008297:Male; D008875:Middle Aged; D010766:Phosphorylation; D011379:Prognosis; D011446:Prospective Studies; D015398:Signal Transduction; D012867:Skin / Q000378:metabolism*; D012878:Skin Neoplasms / Q000378:metabolism* / Q000473:pathology; D055815:Young Adult