Areca nut components affect COX-2, cyclin B1/cdc25C and keratin expression, PGE2 production in keratinocyte is related to reactive oxygen species, CYP1A1, Src, EGFR and Ras signaling.

Abstract

AIMS:

Chewing of betel quid (BQ) increases the risk of oral cancer and oral submucous fibrosis (OSF), possibly by BQ-induced toxicity and induction of inflammatory response in oral mucosa.

METHODS:

Primary gingival keratinocytes (GK cells) were exposed to areca nut (AN) components with/without inhibitors. Cytotoxicity was measured by 3-(4,5-dimethyl- thiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. mRNA and protein expression was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting. PGE2/PGF2α production was measured by enzyme-linked immunosorbent assays.

RESULTS:

Areca nut extract (ANE) stimulated PGE2/PGF2α production, and upregulated the expression of cyclooxygenase-2 (COX-2), cytochrome P450 1A1 (CYP1A1) and hemeoxygenase-1 (HO-1), but inhibited expression of keratin 5/14, cyclinB1 and cdc25C in GK cells. ANE also activated epidermal growth factor receptor (EGFR), Src and Ras signaling pathways. ANE-induced COX-2, keratin 5, keratin 14 and cdc25C expression as well as PGE2 production were differentially regulated by α-naphthoflavone (a CYP 1A1/1A2 inhibitor), PD153035 (EGFR inhibitor), pp2 (Src inhibitor), and manumycin A (a Ras inhibitor). ANE-induced PGE2 production was suppressed by piper betle leaf (PBL) extract and hydroxychavicol (two major BQ components), dicoumarol (a

NAD(P)H:

Quinone Oxidoreductase--NQO1 inhibitor) and curcumin. ANE-induced cytotoxicity was inhibited by catalase and enhanced by dicoumarol, suggesting that AN components may contribute to the pathogenesis of OSF and oral cancer via induction of aberrant differentiation, cytotoxicity, COX-2 expression, and PGE2/PGF2α production.

CONCLUSIONS:

CYP4501A1, reactive oxygen species (ROS), EGFR, Src and Ras signaling pathways could all play a role in ANE-induced pathogenesis of oral cancer. Addition of PBL into BQ and curcumin consumption could inhibit the ANE-induced inflammatory response.

• Authors: Mei-Chi Chang, Yi-Jane Chen, Hsiao-Hua Chang, Chiu-Po Chan, Chien-Yang Yeh, Yin-Lin Wang, Ru-Hsiu Cheng, Liang-Jiunn Hahn, Jiiang-Huei Jeng
• Journal: PloS one (PLoS One) Vol. 9 Issue 7 Pg. e101959 ( 2014) ISSN: 1932-6203 [Electronic] United States
• PMID: 25051199 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• CCNB1 protein, human
• Cyclin B1
• Plant Extracts
• Reactive Oxygen Species
• Keratins
• Dicumarol
• CYP1A1 protein, human
• Cytochrome P-450 CYP1A1
• HMOX1 protein, human
• Heme Oxygenase-1
• Cyclooxygenase 2
• PTGS2 protein, human
• EGFR protein, human
• ErbB Receptors
• src-Family Kinases
• CDC25C protein, human
• cdc25 Phosphatases
• ras Proteins
• Curcumin
• Dinoprostone

Topics

• Areca (chemistry)
• Cells, Cultured
• Curcumin (pharmacology)
• Cyclin B1 (metabolism)
• Cyclooxygenase 2 (genetics, metabolism)
• Cytochrome P-450 CYP1A1 (metabolism)
• Dicumarol (pharmacology)
• Dinoprostone (biosynthesis)
• ErbB Receptors (metabolism)
• Gene Expression (drug effects)
• Gingiva (pathology)
• Heme Oxygenase-1 (metabolism)
• Humans
• Keratinocytes (drug effects, metabolism)
• Keratins (genetics, metabolism)
• Plant Extracts (toxicity)
• Reactive Oxygen Species (metabolism)
• Signal Transduction
• cdc25 Phosphatases (genetics, metabolism)
• ras Proteins (metabolism)
• src-Family Kinases (metabolism)