title: hsa-miR-4516 mediated downregulation of STAT3/CDK6/UBE2N plays a role in PUVA induced apoptosis in keratinocytes.
abstract: Psoriasis is a chronic inflammatory skin disorder mediated by cross-talk occurring between epidermal keratinocytes, dermal vascular cells and immunocytes. Literature reveals that Signal transducer and activator of transcription 3 (STAT3), a protein involved in transmitting extracellular signals to the nucleus, is a possible important link between keratinocytes and immunocytes and is crucial to the development of psoriasis. Although photochemotherapy using UV in combination with 8 methoxypsoralen is one of the most effective therapy for moderate to severe plaque psoriasis, its mechanism of action is largely unknown. Herein, we studied the change in miRNA profiles of cultured human keratinocytes (HaCaT cells) before and after in vitro PUVA treatment by 8 methoxypsoralen and found significant up regulation of hsa-miR-4516. We for the first time demonstrate that ectopic expression of hsa-miR-4516 directly targets STAT3 protein by binding to its 3'UTR in HaCaT cells as confirmed by Luciferase reporter assays and Western blot analysis. We further show that overexpression of hsa-miR-4516 downregulates STAT3, p-STAT3, CDK6, and UBE2N proteins that are consistently upregulated in psoriasis and induces apoptosis in HaCaT cells. We also observed that anti-miR-4516 treatment was able to partially inhibit PUVA-induced apoptosis, suggesting that miR-4516 is involved in PUVA-induced apoptosis. Taken together, these results not only indicate the mechanistic involvement of hsa-miR-4516 in PUVA mediated effects by down-regulating STAT3 in HaCaT keratinocytes, but also highlight the potential of hsa-miR-4516 in development of novel therapeutic strategies. J. Cell. Physiol. 229: 1630-1638, 2014. © 2014 Wiley Periodicals, Inc.
authors.affiliation: Functional Genomics Unit, CSIR, Institute of Genomics and Integrative Biology, Delhi, India.
authors.forename: Shruti
authors.initials: S
authors.lastname: Chowdhari
authors.forename: Neeru
authors.initials: N
authors.lastname: Saini
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D001483:Base Sequence; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D051361:Cyclin-Dependent Kinase 6 / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D020869:Gene Expression Profiling; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology / Q000528:radiation effects; D008730:Methoxsalen / Q000494:pharmacology / Q000627:therapeutic use; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D011701:PUVA Therapy; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D011565:Psoriasis / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D015203:Reproducibility of Results; D050796:STAT3 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics* / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D020413:3' Untranslated Regions; C000591812:MIRN-4516 microRNA, human; D035683:MicroRNAs; D050796:STAT3 Transcription Factor; C471607:UBE2N protein, human; D044763:Ubiquitin-Conjugating Enzymes; D051361:Cyclin-Dependent Kinase 6; D008730:Methoxsalen