The lncRNA VELUCT strongly regulates viability of lung cancer cells despite its extremely low abundance

• Verfasst von: Seiler, Jana [VerfasserIn]
• Verfasst von: Breinig, Marco [VerfasserIn]
• Verfasst von: Caudron-Herger, Maïwen [VerfasserIn]
• Verfasst von: Polycarpou-Schwarz, Maria [VerfasserIn]
• Verfasst von: Boutros, Michael [VerfasserIn]
• Verfasst von: Diederichs, Sven [VerfasserIn]
• Titel: The lncRNA VELUCT strongly regulates viability of lung cancer cells despite its extremely low abundance
• Verf.angabe: Jana Seiler, Marco Breinig, Maïwen Caudron-Herger, Maria Polycarpou-Schwarz, Michael Boutros and Sven Diederichs
• E-Jahr: 2017
• Jahr: 04 February 2017
• Umfang: 12 S.
• Fussnoten: Gesehen am 17.10.2018
• Titel Quelle: Enthalten in: Nucleic acids research
• Ort Quelle: Oxford : Oxford Univ. Press, 1974
• Jahr Quelle: 2017
• Band/Heft Quelle: 45(2017), 9, Seite 5458-5469
• ISSN Quelle: 1362-4962
• DOI: doi:10.1093/nar/gkx076
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1582030634

Abstract

Little is known about the function of most non-coding RNAs (ncRNAs). The majority of long ncRNAs (lncRNAs) is expressed at very low levels and it is a matter of intense debate whether these can be of functional relevance. Here, we identified lncRNAs regulating the viability of lung cancer cells in a highthroughput RNA interference screen. Based on our previous expression profiling, we designed an siRNA library targeting 638 lncRNAs upregulated in human cancer. In a functional siRNA screen analyzing the viability of lung cancer cells, the most prominent hit was a novel lncRNA which we called Viability Enhancing LUng Cancer Transcript (VELUCT). In silico analyses confirmed the non-coding properties of the transcript. Surprisingly, VELUCT was below the detection limit in total RNA from NCI-H460 cells by RT-qPCR as well as RNA-Seq, but was robustly detected in the chromatin-associated RNA fraction. It is an extremely low abundant lncRNA with an RNA copy number of less than one copy per cell. Blocking transcription with actinomycin D revealed that VELUCT RNA was highly unstable which may partially explain its low steady-state concentration. Despite its extremely low abundance, loss-of-function of VELUCT with three independent experimental approaches in three different lung cancer cell lines led to a significant reduction of cell viability: Next to four individual siRNAs, also two complex siPOOLs as well as two antisense oligonucleotides confirmed the strong and specific phenotype.