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| Status |
Public on Feb 09, 2017 |
| Title |
Aberrant methylation-mediated silencing of microRNAs contributes to HPV-induced anchorage independence [miR-Agilent] |
| Organism |
Homo sapiens |
| Experiment type |
Non-coding RNA profiling by array
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| Summary |
Cervical cancer and a subset of anogenital and head-and-neck carcinomas are caused by persistent infection with high-risk types of the human papillomavirus (hrHPV). Early stages of hrHPV-induced carcinogenesis can be faithfully mimicked in vitro. A major hallmark of hrHPV-transformed cells is their ability to grow anchorage independently, an oncogenic trait known to depend on inactivation of tumour suppressor genes. This study used an in vitro model of hrHPV-induced transformation to delineate in a longitudinal manner to what extent DNA methylation-mediated silencing of tumour suppressive microRNAs (miRNAs) contributed to hrHPV-induced anchorage independence. Genome-wide miRNA expression profiles were yielded from anchorage dependent (n=11) and independent passages (n=19) of 4 hrHPV-immortalised keratinocyte cell lines with and without demethylating treatment (DAC). Unsupervised clustering analysis showed that overall miRNA expression patterns discriminated between anchorage dependent and independent cells. Ten miRNA genes potentially silenced by methylation were selected and validated by bisulfite sequencing and methylation-specific PCR. Hsa-mir-129-2, -137, -935, -3663, -3665, and -4281 showed increased methylation in both HPV-transformed keratinocytes and cervical cancer cell lines compared to primary keratinocytes. Mature miRNAs derived from hsa-mir-129-2, -137, -3663, and -3665 decreased anchorage independence in cervical cancer cell lines. Finally, significantly increased methylation of hsa-mir-129-2, -935, -3663, -3665, and -4281 was observed in cervical (pre)cancerous lesions, underlining the clinical relevance of our findings. In conclusion, methylation-mediated silencing of tumour suppressive miRNAs contributes to the acquisition of anchorage independence, supporting the importance of miRNAs during early stages of carcinogenesis and underlining their potential as both disease markers and therapeutic targets.
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| Overall design |
6-8 sequential passages of 4 HPV-transformed human foreskin primary keratinocyte cell lines were treated with the demethylating agent 5-aza-2’-deoxycytidine (DAC). Genome-wide microRNA (miRNA) expression was measured by Agilent 8x60K miRNA microarrays for all 6-8 passages of all 4 cell lines with and without DAC treatment.
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| Contributor(s) |
Wilting SM, Miok V, Jaspers A, Boon D, Sørgård H, Lando M, Snoek BC, van Wieringen WN, Meijer CJ, Lyng H, Snijders PJ, Steenbergen RD |
| Citation(s) |
27270309 |
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| Submission date |
Feb 24, 2016 |
| Last update date |
Feb 09, 2017 |
| Contact name |
Saskia Wilting |
| E-mail(s) |
s.wilting@vumc.nl
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| Organization name |
VU University Medical Center
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| Street address |
De Boelelaan 1117
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| City |
Amsterdam |
| ZIP/Postal code |
1081 HV |
| Country |
Netherlands |
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| Platforms (1) |
| GPL15159 |
Agilent-031181 Unrestricted_Human_miRNA_V16.0_Microarray 030840 (Probe Name version) |
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| Samples (60)
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| This SubSeries is part of SuperSeries: |
| GSE78279 |
Aberrant methylation-mediated silencing of microRNAs contributes to HPV-induced anchorage independence |
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| Relations |
| BioProject |
PRJNA313084 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE78278_RAW.tar |
483.5 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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