A ROS/STAT3/HIF-1α signaling cascade mediates EGF-induced TWIST1 expression and prostate cancer cell invasion.

Abstract

BACKGROUND:

Epidermal growth factor (EGF) has been known to induce epithelial-mesenchymal transition (EMT) and prostate cancer cell progression. However, a detailed underlying mechanism by which EGF induces EMT and prostate cancer cell progression remained to be answered. Hypoxia-inducible factor (HIF)-1α and TWIST1 are transcription factors implicated in EMT and cancer metastasis. The purpose of this study is to determine the underlying mechanism of EGF-induced TWIST1 expression and prostate cancer invasion.

METHODS:

siRNAs were used to silence genes. Immunoblotting, quantitative RT-PCR and immunofluorescence analysis were used to examine protein or mRNA expression. Modified Boyden chamber and invasion assay kit with Matrigel-coated inserts were used to determine prostate cancer cell migration and invasion, respectively.

RESULTS:

We observed that EGF induced HIF-1α expression and morphological change of prostate cancer epithelial cells to mesenchymal cells. Silencing HIF-1α expression dramatically reduced EGF-induced TWIST1 expression and prostate cancer cell EMT. Conversely, transfection of the cells with HIF-1α siRNA reversed the reduced E-cadherin expression by EGF. Pretreatment of the cells with pharmacological inhibitors of reactive oxygen species [ROS, N-acetylcysteine (NAC)] and STAT3 (WP1066) but not p38 MAPK (SB203580) significantly reduced EGF-induced HIF-1α mRNA and protein expression. Further, pretreatment of the cells with NAC attenuated EGF-induced STAT3 phosphorylation. In addition, we showed that TWIST1 mediated EGF-induced N-cadherin expression, leading to prostate cancer invasion.

CONCLUSIONS:

We demonstrate a mechanism by which EGF promotes prostate cancer cell progression through a ROS/STAT3/HIF-1α/TWIST1/N-cadherin signaling cascade, providing novel biomarkers and promising therapeutic targets for prostate cancer cell progression.

• Authors: Kyung Hwa Cho, Moon Jung Choi, Kang Jin Jeong, Jeong Jin Kim, Min Ha Hwang, Shang Cheul Shin, Chang Gyo Park, Hoi Young Lee
• Journal: The Prostate (Prostate) Vol. 74 Issue 5 Pg. 528-36 (May 2014) ISSN: 1097-0045 [Electronic] United States
• PMID: 24435707 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 Wiley Periodicals, Inc.

Chemical References

• Cadherins
• HIF1A protein, human
• Hypoxia-Inducible Factor 1, alpha Subunit
• Nuclear Proteins
• RNA, Small Interfering
• Reactive Oxygen Species
• STAT3 Transcription Factor
• TWIST1 protein, human
• Twist-Related Protein 1
• Epidermal Growth Factor
• Acetylcysteine

Topics

• Acetylcysteine (pharmacology)
• Cadherins (genetics, metabolism)
• Cell Line, Tumor
• Cell Movement (drug effects, physiology)
• Disease Progression
• Epidermal Growth Factor (pharmacology)
• Epithelial-Mesenchymal Transition
• Gene Expression Regulation, Neoplastic
• Gene Silencing
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit (genetics, metabolism)
• Male
• Neoplasm Invasiveness (pathology)
• Nuclear Proteins (genetics, metabolism)
• Phosphorylation (drug effects, physiology)
• Prostatic Neoplasms (metabolism, pathology)
• RNA, Small Interfering
• Reactive Oxygen Species (metabolism)
• STAT3 Transcription Factor (genetics, metabolism)
• Signal Transduction (drug effects, physiology)
• Twist-Related Protein 1 (genetics, metabolism)