title: Human dermal fibroblasts HDFa can be used as an appropriate healthy control for PMMA nanoparticles-survivin molecular beacon cellular uptake studies.
abstract: Survivin is a member of the inhibitor of apoptosis protein family (IAPs); besides its inhibitory action on apoptosis, it is also involved in the regulation of cell division. The protein expression is up-regulated in several cancers, being involved in the tumor progression and evasion of apoptosis. In line with its physiological roles, it is expressed also in several healthy tissues. The high expression of survivin in cancer cells correlates to poor prognosis and resistance to chemotherapeutic treatment, thus making this protein an attractive target in anticancer therapy. The dual role of survivin in cells, regulation of cell division and inhibition of apoptosis, combined with controversial data concerning the expression in normal tissues, emphasize the need to have an appropriate healthy control for in vitro studies. Aim of this study is to highlight this problem and to clarify the experimental conditions in which HDFa fibroblasts represent a negative control for survivin mRNA expression while ensuring the NPs-MB uptake. In this paper, by using confocal microscopy analysis supported by RT- and real-time-PCR experiments studies, we showed that HDFa fibroblasts represent a healthy negative control for survivin mRNA expression, only at very low cell density or at confluence. At the same time, we demonstrated that HDFa at any cell density are able to internalize NPs-MB after 6h of treatment.
authors: Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino, Italy. Electronic address: b.adinolfi@ifac.cnr.it.
authors: Barbara
authors: B
authors: Adinolfi
authors: Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Università di Pisa, Via S. Zeno 31, 56127 Pisa, Italy. Electronic address: marpell@dfb.unipi.it.
authors: Mario
authors: M
authors: Pellegrino
authors: Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino, Italy. Electronic address: f.baldini@ifac.cnr.it.
authors: Francesco
authors: F
authors: Baldini
mesh_terms: D000328:Adult; D002470:Cell Survival; D020405:Dermis / Q000166:cytology*; D004705:Endocytosis; D005347:Fibroblasts; D005786:Gene Expression Regulation; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism*; D015335:Molecular Probes / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction; D000077022:Survivin
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D015335:Molecular Probes; D000077022:Survivin; D019904:Polymethyl Methacrylate
keywords: Cellular uptake; Fibroblasts HDFa; Healthy control; Molecular beacon (MB); Polymethylmetacrilate nanoparticles (PMMA-NPs); Survivin expression