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Near‐genomewide RNA i screening for regulators of BRAF V 600E ‐induced senescence identifies RASEF , a gene epigenetically silenced in melanoma
Author(s) -
Kaplon Joanna,
HömigHölzel Cornelia,
Gao Linda,
Meissl Katrin,
Verdegaal Els M. E.,
Burg Sjoerd H.,
Doorn Remco,
Peeper Daniel S.
Publication year - 2014
Publication title -
pigment cell and melanoma research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.618
H-Index - 105
eISSN - 1755-148X
pISSN - 1755-1471
DOI - 10.1111/pcmr.12248
Subject(s) - senescence , biology , small hairpin rna , cancer research , context (archaeology) , melanoma , rna , pten , dna methylation , gene , suppressor , microbiology and biotechnology , gene expression , genetics , apoptosis , pi3k/akt/mtor pathway , paleontology
Summary The activation of oncogenes in primary cells blocks proliferation by inducing oncogene‐induced senescence ( OIS ), a highly potent in vivo tumor‐suppressing program. A prime example is mutant BRAF , which drives OIS in melanocytic nevi. Progression to melanoma occurs only in the context of additional alteration(s) like the suppression of PTEN , which abrogates OIS . Here, we performed a near‐genomewide short hairpin (sh) RNA screen for novel OIS regulators and identified by next generation sequencing and functional validation seven genes. While all but one were upregulated in OIS , depletion of each of them abrogated BRAF V 600E ‐induced arrest. With genome‐wide DNA methylation analysis, we found one of these genes, RASEF , to be hypermethylated in primary cutaneous melanomas but not nevi. Bypass of OIS by depletion of RASEF was associated with suppression of several senescence biomarkers including senescence‐associated ( SA )‐ β ‐galactosidase activity, interleukins, and tumor suppressor p15 INK 4B . Restoration of RASEF expression inhibited proliferation. These results illustrate the power of sh RNA OIS bypass screens and identify a potential novel melanoma suppressor gene.