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Design of a miRNA sponge for the miR-17 miRNA family as a therapeutic strategy against vulvar carcinoma
- Source :
- Molecular and Cellular Probes. 29:420-426
- Publication Year :
- 2015
- Publisher :
- Elsevier BV, 2015.
-
Abstract
- Dysregulation of microRNAs has been studied thoroughly, and has been observed in a variety of tumors including vulvar carcinomas, a rare type of gynecological tumor with increasing incidence. However, very few therapeutic alternatives have reached the clinical setting, and there is an urgent unmet need to develop novel strategies for patients with this tumor type. Thus, a microRNA (miRNA) sponge for the miR-17 miRNA family was designed, synthesized and validated in vitro in order to explore a new therapeutic strategy based on inhibiting this oncogenic miRNA family in vulvar cancer. Members of the miR-17 family were evaluated for expression in a vulvar tumor cell line (SW954) and 20 HPV negative formalin-fixed paraffin-embedded (FFPE) samples by quantitative real-time PCR (qRT-PCR). Six in tandem, bulged sequences that were complementary to these miRNAs were designed, synthesized, cloned, and transfected into SW954 cells. A luciferase reporter assay with a psiCheck2 vector was used to test the specificity of the sponge sequences for miR-17 family miRNA binding. Taqman qRT-PCR was used to test how the sponges affected miRNA expression. In FFPE samples, higher expression of miR-20a and miR-106a correlated with deeper tumor invasion (P = 0.0187 and P = 0.0404, respectively). The luciferase reporter assay validated the specificity of the sponge for miR-17 family members. Using qRT-PCR, we confirmed this specificity with decreased expression in 5 (out of six) miRNAs of the miR-17 family in SW954 cells. Although our results are preliminary, these results demonstrate that these miRNA sponges are potent inhibitors of the miR-17 family of miRNAs in SW954. Therefore, this miRNA-specific sponge may be developed into a novel therapeutic treatment for patients with vulvar cancer.
- Subjects :
- Vulvar Neoplasms
Gene Expression Profiling
MiRNA binding
Cell Biology
Transfection
In Vitro Techniques
Vulvar cancer
Biology
medicine.disease
Bioinformatics
Gene Expression Regulation, Neoplastic
MicroRNAs
Cell culture
Cell Line, Tumor
Multigene Family
microRNA
Cancer research
medicine
TaqMan
Humans
Female
Molecular Targeted Therapy
Vector (molecular biology)
Vulvar Carcinoma
Molecular Biology
Subjects
Details
- ISSN :
- 08908508
- Volume :
- 29
- Database :
- OpenAIRE
- Journal :
- Molecular and Cellular Probes
- Accession number :
- edsair.doi.dedup.....ec5d789872675831e3d2cdbb072db065