Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

MicroRNA sequencing and Novel HPV miRNA discovery in HPV cell lines and human tissue samples

ABSTRACT: Human papillomaviruses (HPV) preferentially infect keratinocytes of mucous membranes or skin and cause numerous benign and malignant lesions at different anatomical locations. A number of DNA viruses encode their own miRNAs as well. To date, no studies have been able to validate viral miRNAs in papillomavirus infected cells using standard sequencing or next generation sequencing techniques. We sequenced small RNA (sRNA) libraries of two HPV 16 immortalized cell lines, HPK IA and HPK II, and ten formalin fixed paraffin embedded (FFPE) tissue samples from HPV infected cervical epithelium by SOLiD 4 technology. We used the miRSeqNovel software, to predict novel miRNAs and their likely pre-miRNAs. We further validated (qRT-PCR, northern blotting and in situ hybridization) the candidate miRNAs in a number of tissue samples from HPV associated cervical disease and also in HPV 16 positive cell lines CaSki and SiHa. Altogether nine candidate microRNAs were identified. The expression of four out of five studied miRNAs was confirmed in human tissue or human epithelial cell lines harboring HPV 16. Small RNA (sRNA) libraries of two HPV 16 immortalized cell lines, HPK IA and HPK II, and ten formalin fixed paraffin embedded (FFPE) tissue samples from HPV infected cervical epithelium, were made according to SOLiD sequencing instructions.

ORGANISM(S): Homo sapiens

SUBMITTER: Kui Qian

PROVIDER: E-GEOD-42380 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Identification and validation of human papillomavirus encoded microRNAs.

Qian Kui K Pietilä Tuuli T Rönty Mikko M Michon Frederic F Frilander Mikko J MJ Ritari Jarmo J Tarkkanen Jussi J Paulín Lars L Auvinen Petri P Auvinen Eeva E

PloS one 20130730 7

We report here identification and validation of the first papillomavirus encoded microRNAs expressed in human cervical lesions and cell lines. We established small RNA libraries from ten human papillomavirus associated cervical lesions including cancer and two human papillomavirus harboring cell lines. These libraries were sequenced using SOLiD 4 technology. We used the sequencing data to predict putative viral microRNAs and discovered nine putative papillomavirus encoded microRNAs. Validation w ...[more]

PMID: 23936163

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Project description:In most cases human papillomavirus (HPV) infections are cleared from the cervical cells by the immune system itself, but in a few cases, where there is persistent HPV infection, it can lead to cervical intraepithelial neoplasia (CIN) progression and ultimately invasive cervical carcinoma. The cytopathic effect is in general accompanied by chronic inflammation, which produces inflammation cytokines that contribute to DNA damage, and at the same time, aberrations occurred in the host DNA repair mechanisms, thus lead to HPV genomic integration into the host cells which propels cell immortalization. In this study, we reported the genome-wide expression profiles of both microRNAs (miRNAs) and mRNAs from 24 cervical samples with consecutive stages of normal, CIN I (mild dysplasia) and CIN III (severe dysplasia and carcinoma in situ), and presented the SIG++ algorithm which is founded on the evolution process of intermolecular regulation change during disease progression, to identify the significant change of miRNA-mRNA regulations rather than the expression change, across different disease stages, thereupon elucidating the molecular mechanisms of increasing host genomic instability as disease progresses. As reconstructing miRNA differential networks, we found that at each stage of CIN, there respectively exists specific miRNA regulations mediating chronic inflammation persistence, genome instability and cell survival, which coordinately carrys out the integration of HPV genomes into the host cell genomes, and finally results in cell immortalization. Beyond the specific implications for cervical carcinogenesis, this work establishes a new framework for studying the biology of miRNAs in pathogenesis from the perspective of miRNA differential regulation, and helps ensure the comprehensiveness of miRNA-mediated genetic regulatory pathways. There are totally 24 clinical samples in this study comprises three stages: 7 normal cervix samples (HPV-), 9 CIN I samples (HPV+) and CIN III samples (HPV+), where normal refers to the adjacent tissue of early lesions. For each sample, its total RNA was extracted and purified, then separately hybridized to Illumina HumanHT-12 V4.0 expression beadchip (gene symbol) and Illumina Human v2 MicroRNA Expression BeadChip, for examining the expression profiles of mRNAs and miRNAs, respectively.

Dataset Information

MicroRNA sequencing and Novel HPV miRNA discovery in HPV cell lines and human tissue samples

Publications

Identification and validation of human papillomavirus encoded microRNAs.

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