Dicer Sequencing, Whole Genome Methylation Profiling, mRNA and smallRNA Sequencing Analysis in Basal Cell Carcinoma.


Journal

Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
ISSN: 1421-9778
Titre abrégé: Cell Physiol Biochem
Pays: Germany
ID NLM: 9113221

Informations de publication

Date de publication:
2019
Historique:
accepted: 22 10 2019
entrez: 25 10 2019
pubmed: 28 10 2019
medline: 26 11 2019
Statut: ppublish

Résumé

Perturbations in the expression of microRNAs (miRNAs) and their maturing machinery components such as Dicer have been previously described for basal cell carcinoma (BCC). However, the mutational status of Dicer in BCC is unclear. Further, the sclerodermiform subtype of BCC (sBCC) has not been previously investigated regarding its methylation profile or its smallRNA expression profile via RNA sequencing. We conducted this study to investigate the mutational status of Dicer in BCC. Dicer sequencing was performed on the Illumina MiSeq System in a total of 16 BCC samples (8 nodular BCCs, 8 sBCCs) and mapped against the human reference genome (i.e., hg19). Dicer sequencing was performed in all 16 BCC samples. We performed whole genome methylation profiling with Infinium MethylationEPIC BeadChips as well as mRNA and smallRNA sequencing in 5 sBCCs with the Illumina NextSeq500 next-generation sequencing system. Compared to the wildtype Dicer sequence, we found 5 to 7 variants per sBCC sample including insertion, deletion, and multiple nucleotide variants. Global methylation profiles were highly similar between groups. mRNA sequencing revealed S100A9, KRT14, KRT10, S100A8, S100A7, COX1, KRT1, COX3, and smallRNA sequencing analysis miR-21, miR-99a, miR26-a-2, let-7f, let-7g, let-7i, miR-100, and miR-205 were the most strongly expressed in sBCCs. We identified a variety of Dicer mutations that could play a role in aberrant miRNA expression in BCC. The noted RNA sequences should be further evaluated in functional studies to explore their potential pathogenetic role in sBCC.

Sections du résumé

BACKGROUND/AIMS OBJECTIVE
Perturbations in the expression of microRNAs (miRNAs) and their maturing machinery components such as Dicer have been previously described for basal cell carcinoma (BCC). However, the mutational status of Dicer in BCC is unclear. Further, the sclerodermiform subtype of BCC (sBCC) has not been previously investigated regarding its methylation profile or its smallRNA expression profile via RNA sequencing. We conducted this study to investigate the mutational status of Dicer in BCC.
METHODS METHODS
Dicer sequencing was performed on the Illumina MiSeq System in a total of 16 BCC samples (8 nodular BCCs, 8 sBCCs) and mapped against the human reference genome (i.e., hg19). Dicer sequencing was performed in all 16 BCC samples. We performed whole genome methylation profiling with Infinium MethylationEPIC BeadChips as well as mRNA and smallRNA sequencing in 5 sBCCs with the Illumina NextSeq500 next-generation sequencing system.
RESULTS RESULTS
Compared to the wildtype Dicer sequence, we found 5 to 7 variants per sBCC sample including insertion, deletion, and multiple nucleotide variants. Global methylation profiles were highly similar between groups. mRNA sequencing revealed S100A9, KRT14, KRT10, S100A8, S100A7, COX1, KRT1, COX3, and smallRNA sequencing analysis miR-21, miR-99a, miR26-a-2, let-7f, let-7g, let-7i, miR-100, and miR-205 were the most strongly expressed in sBCCs.
CONCLUSION CONCLUSIONS
We identified a variety of Dicer mutations that could play a role in aberrant miRNA expression in BCC. The noted RNA sequences should be further evaluated in functional studies to explore their potential pathogenetic role in sBCC.

Identifiants

pubmed: 31647206
doi: 10.33594/000000171
doi:

Substances chimiques

MicroRNAs 0
RNA, Messenger 0
DICER1 protein, human EC 3.1.26.3
Ribonuclease III EC 3.1.26.3
DEAD-box RNA Helicases EC 3.6.4.13

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

760-773

Subventions

Organisme : Brigitte and Dr. Konstanze Wegener-Stiftung
Pays : Germany

Informations de copyright

© Copyright by the Author(s). Published by Cell Physiol Biochem Press.

Déclaration de conflit d'intérêts

All authors hereby disclose any commercial associations that may pose or create a conflict of interest with the information presented in this manuscript. The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Auteurs

Michael Sand (M)

Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, Bochum, Germany.
Department of Plastic Surgery, St. Josef Hospital, Catholic Clinics of the Ruhr Peninsula, Essen, Germany, michael.sand@ruhr-uni-bochum.de.

Annabelle Bromba (A)

Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, Bochum, Germany.

Daniel Sand (D)

Department of Physiological Science, University of California Los Angeles (UCLA), Los Angeles, CA, USA.

Thilo Gambichler (T)

Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, Bochum, Germany.

Schapoor Hessam (S)

Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, Bochum, Germany.

Jürgen C Becker (JC)

Department of Translational Skin Cancer Research, University Hospital Essen, Essen, Germany.
German Cancer Consortium (DKTK), Essen, Germany.
German Cancer Research Center (DKFZ), Heidelberg, Germany.

Eggert Stockfleth (E)

Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, Bochum, Germany.

Thomas Meyer (T)

Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, Bochum, Germany.

Falk G Bechara (FG)

Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, Bochum, Germany.

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Classifications MeSH