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Comprehensive analysis of alterations in the miRNome in response to photodynamic treatment
Doris Bach1, Julia Fuereder, Michael Karbiener
1Department of Internal Medicine I, Paracelsus Medical University, Salzburg, Austria. d.bach@salk.at
Abstract:
Photodynamic therapy (PDT) is a local tumour treatment accepted for a number of indications. PDT operates via the cellular stress response through the production of reactive oxygen species and subsequent cellular damage, resulting in cell death. Although PDT-induced signalling and cytotoxicity mechanisms have been investigated, the effect of PDT on microRNA (miRNA) expression is largely unknown. Therefore, we conducted a comprehensive microarray-based analysis of the miRNome of human epidermoid carcinoma cells (A431) following in vitro photodynamic treatment using polyvinylpyrrolidone hypericin (PVPH) as a photosensitiser and nearly homogeneous apoptosis-inducing conditions. Using microarray analysis we found eight miRNAs to be significantly differentially expressed 5h post treatment compared with the baseline levels and three miRNAs with more than 2-fold differential expression that could be detected in 1 or 2 biological replicates. The verification of these results by quantitative RT-PCR including a detailed time-course revealed an up to 15-fold transient over-expression of miR-634, miR-1246, miR-1290 and miR-487b compared with the basal level. For these miRNAs, in silico mRNA target prediction yielded numerous target transcripts involved in the regulation of cell stress, apoptosis, cell adherence and proliferation. This study provides the first comprehensive miRNome analysis after PDT treatment and may help to develop novel miRNA-based therapeutic approaches to further increase the efficiency of PDT.
Insights
Photodynamic therapy (PDT) alters microRNA (miRNA) expression in cancer cells. This study reveals specific miRNA changes after PDT, offering potential for new miRNA-based cancer treatments.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Photodynamic therapy (PDT) is an established local cancer treatment.
- PDT induces cell death via reactive oxygen species and cellular stress.
- The impact of PDT on microRNA (miRNA) expression remains largely uncharacterized.
Purpose of the Study:
- To comprehensively analyze the miRNome of human epidermoid carcinoma cells (A431) following in vitro PDT.
- To identify specific microRNAs (miRNAs) affected by PDT treatment.
- To explore potential novel miRNA-based therapeutic strategies for enhancing PDT efficacy.
Main Methods:
- Microarray analysis of the miRNome in A431 cells after in vitro PDT using polyvinylpyrrolidone hypericin (PVPH).
- Quantitative RT-PCR for verification and detailed time-course analysis of differentially expressed miRNAs.
- In silico mRNA target prediction for identified miRNAs.
Main Results:
- Eight miRNAs showed significant differential expression 5 hours post-PDT.
- Three miRNAs exhibited more than 2-fold differential expression.
- Quantitative RT-PCR confirmed transient overexpression (up to 15-fold) of miR-634, miR-1246, miR-1290, and miR-487b.
- In silico analysis predicted numerous mRNA targets involved in cell stress, apoptosis, adherence, and proliferation.
Conclusions:
- This is the first comprehensive miRNome analysis following PDT.
- PDT significantly alters miRNA expression profiles in cancer cells.
- The identified miRNAs and their targets may offer novel therapeutic targets to improve PDT outcomes.
