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Characterization of pancreatic ductal adenocarcinoma using whole transcriptome sequencing and copy number analysis by single-nucleotide polymorphism array

  • Authors:
    • Mariacristina Di Marco
    • Annalisa Astolfi
    • Elisa Grassi
    • Silvia Vecchiarelli
    • Marina Macchini
    • Valentina Indio
    • Riccardo Casadei
    • Claudio Ricci
    • Marielda D'Ambra
    • Giovanni Taffurelli
    • Carla Serra
    • Giorgio Ercolani
    • Donatella Santini
    • Antonia D'Errico
    • Antonio Daniele Pinna
    • Francesco Minni
    • Sandra Durante
    • Laura Raffaella Martella
    • Guido Biasco

  • View Affiliations

  • Published online on: September 22, 2015     https://doi.org/10.3892/mmr.2015.4344
  • Pages: 7479-7484
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Abstract

The aim of the current study was to implement whole transcriptome massively parallel sequencing (RNASeq) and copy number analysis to investigate the molecular biology of pancreatic ductal adenocarcinoma (PDAC). Samples from 16 patients with PDAC were collected by ultrasound‑guided biopsy or from surgical specimens for DNA and RNA extraction. All samples were analyzed by RNASeq performed at 75x2 base pairs on a HiScanSQ Illumina platform. Single‑nucleotide variants (SNVs) were detected with SNVMix and filtered on dbSNP, 1000 Genomes and Cosmic. Non‑synonymous SNVs were analyzed with SNPs&GO and PROVEAN. A total of 13 samples were analyzed by high resolution copy number analysis on an Affymetrix SNP array 6.0. RNAseq resulted in an average of 264 coding non‑synonymous novel SNVs (ranging from 146‑374) and 16 novel insertions or deletions (In/Dels) (ranging from 6‑24) for each sample, of which a mean of 11.2% were disease‑associated and somatic events, while 34.7% were frameshift somatic In/Dels. From this analysis, alterations in the known oncogenes associated with PDAC were observed, including Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations (93.7%) and inactivation of cyclin‑dependent kinase inhibitor 2A (CDKN2A) (50%), mothers against decapentaplegic homolog 4 (SMAD4) (50%), and tumor protein 53 (TP53) (56%). One case that was negative for KRAS exhibited a G13D neuroblastoma RAS viral oncogene homolog mutation. In addition, gene fusions were detected in 10 samples for a total of 23 different intra‑ or inter‑chromosomal rearrangements, however, a recurrent fusion transcript remains to be identified. SNP arrays identified macroscopic and cryptic cytogenetic alterations in 85% of patients. Gains were observed in the chromosome arms 6p, 12p, 18q and 19q which contain KRAS, GATA binding protein 6, protein kinase B and cyclin D3. Deletions were identified on chromosome arms 1p, 9p, 6p, 18q, 10q, 15q, 17p, 21q and 19q which involve TP53, CDKN2A/B, SMAD4, runt‑related transcription factor 2, AT‑rich interactive domain‑containing protein 1A, phosphatase and tensin homolog and serine/threonine kinase 11. In conclusion, genetic alterations in PDCA were observed to involve numerous pathways including cell migration, transforming growth factor‑β signaling, apoptosis, cell proliferation and DNA damage repair. However, signaling alterations were not observed in all tumors and key mutations appeared to differ between PDAC cases.
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November-2015
Volume 12 Issue 5

Print ISSN: 1791-2997
Online ISSN:1791-3004

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  • Journal Metrics
  • Impact Factor: 3.4
  • Ranked Medicine Research and Experimental
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  • CiteScore: 6.9
  • CiteScore Rank: Q2: #98/328 Biochemistry, Genetics and Molecular Biology
  • Source Normalized Impact per Paper (SNIP): 0.721
  • SCImago Journal Rank (SJR): 0.65
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Spandidos Publications style
Di Marco M, Astolfi A, Grassi E, Vecchiarelli S, Macchini M, Indio V, Casadei R, Ricci C, D'Ambra M, Taffurelli G, Taffurelli G, et al: Characterization of pancreatic ductal adenocarcinoma using whole transcriptome sequencing and copy number analysis by single-nucleotide polymorphism array. Mol Med Rep 12: 7479-7484, 2015
APA
Di Marco, M., Astolfi, A., Grassi, E., Vecchiarelli, S., Macchini, M., Indio, V. ... Biasco, G. (2015). Characterization of pancreatic ductal adenocarcinoma using whole transcriptome sequencing and copy number analysis by single-nucleotide polymorphism array. Molecular Medicine Reports, 12, 7479-7484. https://doi.org/10.3892/mmr.2015.4344
MLA
Di Marco, M., Astolfi, A., Grassi, E., Vecchiarelli, S., Macchini, M., Indio, V., Casadei, R., Ricci, C., D'Ambra, M., Taffurelli, G., Serra, C., Ercolani, G., Santini, D., D'Errico, A., Pinna, A. D., Minni, F., Durante, S., Martella, L. R., Biasco, G."Characterization of pancreatic ductal adenocarcinoma using whole transcriptome sequencing and copy number analysis by single-nucleotide polymorphism array". Molecular Medicine Reports 12.5 (2015): 7479-7484.
Chicago
Di Marco, M., Astolfi, A., Grassi, E., Vecchiarelli, S., Macchini, M., Indio, V., Casadei, R., Ricci, C., D'Ambra, M., Taffurelli, G., Serra, C., Ercolani, G., Santini, D., D'Errico, A., Pinna, A. D., Minni, F., Durante, S., Martella, L. R., Biasco, G."Characterization of pancreatic ductal adenocarcinoma using whole transcriptome sequencing and copy number analysis by single-nucleotide polymorphism array". Molecular Medicine Reports 12, no. 5 (2015): 7479-7484. https://doi.org/10.3892/mmr.2015.4344