Abstract
The long non-coding RNAs (lncRNAs) are matter of intense investigation as potential regulators of gene expression. In the case of the transglutaminase 2 gene (TGM2) the databases of genome sequence indicate location of a lncRNA (LOC107987281) within the first intron. This lncRNA is 1000 bp long, arises from 2 exons and starts few nucleotides 3′ of the first splicing site of translated TGM2. We have analysed correlations between expression of LOC107987281 lncRNA and TGM2 mRNA by real-time PCR in K562 cell line untreated or treated with the anticancer drugs TPA (12-O-tetradecanoylphorbol-13-acetate), Docetaxel and Doxorubicin. In the treated cells the lncRNA increase follows the trend of TGM2 transcript. To validate this finding we used HumanExon1_0ST Affymetrix; chip data were background-adjusted, quantile-normalized and summarized using robust multi-array average analysis implemented in the R package. The probesets recognize sequences inside each exon, near intronic splicing sites and others located in the untranslated regions of TGM2 gene. The analysis of total RNA samples in GEO datasets from K562, HL-60, THP-1 and U937 cell lines, untreated or treated with TPA in replicated experiments confirmed our earlier results. These demonstrate correlation between LOC107987281 and TGM2 mRNA in the cell lines (K562, HL60 and THP-1) where increased levels of TGM2 mRNA are produced. Additional array study on 358 samples of several normal and paired tumor tissues leads to the same conclusions, indicating a correlation between full-length TGM2 mRNA and LOC107987281 lncRNA in relation to the development of several tumors.
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Abbreviations
- AD:
-
Alzheimer’s disease
- ATRA:
-
All-trans-retinoic acid
- DMSO:
-
Dimethyl sulfoxide
- GEO:
-
Gene Expression Omnibus
- HD:
-
Huntington’s disease
- HPRT1:
-
Hypoxanthine phosphoribosyltransferase
- lncRNA:
-
Long non-coding RNA
- miRNA:
-
microRNA
- NI:
-
Normalized intensity
- PLCδ1:
-
Phospholipase Cδ1
- RMA:
-
Robust multi-array average
- RT-qPCR:
-
Reverse Transcription- quantitative Polymerase Chain Reaction
- SI:
-
Splicing index
- SPSS:
-
Statistical package for social science
- TPA:
-
12-O-Tetradecanoylphorbol-13-acetate
- TGH:
-
Transglutaminase 2 isoform AA1-548
- TGH2:
-
Transglutaminase 2 isoform AA1-349
- TGM2 :
-
Transglutaminase 2 gene
- tTGV1 :
-
tTG alternative splicing variant 1
- tTGV2 :
-
tTG alternative splicing variant 2
- Tg2:
-
Type 2 transglutaminase
- NT:
-
Untreated
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Acknowledgements
S.V. is supported by Associazione Italiana Ricerca sul Cancro AIRC (IG 17063); CMB is supported by FAR2017 (Unife).
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LM carry out bioinformatic analysis, FB performed the experiments on culture cell lines, MG revised the manuscript critically and suggested some idea to improve the work, NB conceived the study. SV and CMB participated in financing and experimental design. NB, CMB, LM and SV carried it out and drafted the manuscript. All authors approved the final manuscript.
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The research has been performed on commercially available cell lines and taken advantage of data sets store in public databanks not involving Human Participants and/or Animals.
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Suppl. Figure
1 Pattern of evolution divergence of LOC107987281 lncRNA. Cladogram from UCSC Genome Browser (http://genome.UCSC.edu). (TIFF 118 kb)
Suppl. Figure
2 Sequence alignment of LOC107987281 lncRNA. This track shows multiple alignments of 100 vertebrate species in UCSC Genome Browser (http://genome.UCSC.edu). (TIFF 799 kb)
Suppl. Figure
3 Sequence alignment of TGM2. This track shows multiple alignments of 100 vertebrate species in UCSC Genome Browser (http://genome.UCSC.edu). (TIFF 969 kb)
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Minotti, L., Baldassari, F., Galasso, M. et al. A long non-coding RNA inside the type 2 transglutaminase gene tightly correlates with the expression of its transcriptional variants. Amino Acids 50, 421–438 (2018). https://doi.org/10.1007/s00726-017-2528-9
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DOI: https://doi.org/10.1007/s00726-017-2528-9