Silencing of reversion-inducing cysteine-rich protein with Kazal motifs stimulates hyperplastic phenotypes through activation of epidermal growth factor receptor and hypoxia-inducible factor-2a

Authors:
Lee YM1, Lee SH1, Lee KB2, Nguyen MP1, Lee MY3, Park GH3, Kwon MJ
In:
Source: PLoS ONE
Publication Date: (2013)
Issue: 8(12): e84520
Research Area:
Basic Research
Cells used in publication:
Epithelial, prostate (PrEC), human
Species: human
Tissue Origin: prostate
Abstract
Reversion-inducing cysteine-rich protein with Kazal motifs (RECK, a tumor suppressor) is down-regulated by the oncogenic signals and hypoxia, but the biological function of RECK in early tumorigenic hyperplastic phenotypes is largely unknown. Knockdown of RECK by small interfering RNA (siRECK) or hypoxia significantly promoted cell proliferation in various normal epithelial cells. Hypoxia as well as knockdown of RECK by siRNA increased the cell cycle progression, the levels of cyclin D1 and c-Myc, and the phosphorylation of Rb protein (p-pRb), but decreased the expression of p21(cip1), p27(kip1), and p16(ink4A). HIF-2a was upregulated by knockdown of RECK, indicating HIF-2a is a downstream target of RECK. As knockdown of RECK induced the activation of epidermal growth factor receptor (EGFR) and treatment of an EGFR kinase inhibitor, gefitinib, suppressed HIF-2a expression induced by the silencing of RECK, we can suggest that the RECK silenicng-EGFR-HIF-2a axis might be a key molecular mechanism to induce hyperplastic phenotype of epithelial cells. It was also found that shRNA of RECK induced larger and more numerous colonies than control cells in an anchorage-independent colony formation assay. Using a xenograft assay, epithelial cells with stably transfected with shRNA of RECK formed a solid mass earlier and larger than those with control cells in nude mice. In conclusion, the suppression of RECK may promote the development of early tumorigenic hyperplastic characteristics in hypoxic stress.