Naslov
Function of the long noncoding RNA, WNT1-inducible signaling pathway protein 1-overlapping transcript 2 (WISP1-OT2) in human renal cells

Autori
Mirjana Polović, Gerald Schwerdt, Michael Gekle

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
22nd Annual Meeting of the RNA Society

Mjesto i datum
Prag, Češka Republika, 30.05.-03.06. 2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
OTA, ochratoxin A, lncRNA, WISP1

Sažetak
Background: The alternative transcript WISP1-OT2 (WNT1 inducible signaling pathway protein 1-overlapping transcript 2) was discovered upon treatment of primary culture of renal proximal tubule epithelial cells (RPTEC) with the ubiquitous nephrotoxic mycotoxin, ochratoxin A (OTA). WISP1-OT2 is a long noncoding RNA, 2922 nucleotides long, and transcribed from the end of WISP1 gene. We were able to detect WISP1-OT2 in healthy renal tissue and we observed an upregulation in clear cell renal cell carcinoma. WISP1-OT2 is located predominantly in the nucleus ; however, its function is unknown. Aims: (i) Determine the orientation of WISP1-OT2 ; (ii) investigate possible functional link between long noncoding RNA and mRNA of WISP1 gene ; (iii) evaluate the influence of WISP1-OT2 knockdown on the cell function. Methods: Orientation of WISP1-OT2 was investigated by gene-specific reverse transcription and subsequent PCR. To test an inhibitory effect of WISP1-OT2 on WISP1 mRNA, HEK293T cells were treated with PMA (Phorbol myristate acetate) or OTA alone, or in combination, in the presence or absence of antisense LNA™ GapmeRs (hybrid DNA:RNA oligonucleotides) that enter the nucleus and degrade target RNA, i.e. WISP1-OT2. Subsequently, WISP1-OT2 and WISP1 mRNA expression was detected by RT-PCR. To investigate the importance of WISP1-OT2 for gene expression, RNA-seq data were generated in the absence and presence of antisense oligonucleotides against WISP1-OT2. To obtain cellular glycolytic flux, we measured glucose consumption and lactate production. The impact on cell death was determined by caspase-III activation and LDH release. Results: WISP1-OT2 is antisense-oriented long noncoding RNA. PMA induces specifically WISP1 mRNA, whereas OTA induces specifically WISP1-OT2. In the presence of PMA and OTA, WISP1 mRNA is significantly reduced when compared to PMA effect alone. This inhibitory effect of OTA on the WISP1 mRNA is not mediated by WISP1-OT2. WISP1-OT2 modulates cellular energy metabolism ; in its absence cells shift from the TCA cycle to aerobic glycolysis. WISP1-OT2 acts anti-apoptotic, possibly via mitochondrial protection. Additionally, it upregulates growth-arrest specific-6 (Gas6) gene, implicated in cell survival. Conclusions: (i) WISP1-OT2 is the long noncoding RNA, transcribed in antisense orientation relative to WISP1 gene ; (ii) WISP1-OT2 does not influence the expression of WISP1 mRNA ; (iii) WISP1-OT2 acts anti-apoptoti

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA