Research Papers:
Sonoporation by microbubbles as gene therapy approach against liver cancer
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Abstract
Luca Rinaldi1, Veronica Folliero2, Luciana Palomba2, Carla Zannella2, Rachele Isticato3, Raffaele Di Francia4, Massimiliano Berretta5, Ilario de Sio6, Luigi E. Adinolfi1, Giancarlo Morelli7, Secondo Lastoria8, Lucia Altucci6, Carlo Pedone7, Massimiliano Galdiero2 and Gianluigi Franci2
1Department of Medical, Surgical, Neurological, Metabolic and Aging Science, University of Campania “Luigi Vanvitelli”, Naples, Italy
2Department of Experimental Medicine, University of Campania “Luigi Vanvitelli”, Naples, Italy
3Department of Biology, Federico II University, Naples, Italy
4Department of Hematology, National Cancer Institute, Foundation G. Pascale IRCCS, Naples, Italy
5Department of Medical Oncology, National Cancer Institute IRCCS, Aviano, Italy
6Department of Precision Medicine, University of Campania “Luigi Vanvitelli”, Naples, Italy
7Department of Pharmacology, Federico II University, Naples, Italy
8Department of Diagnostic Imaging, Radiation and Metabolic Therapy, National Cancer Institute, Foundation G. Pascale IRCCS, Naples, Italy
Correspondence to:
Luca Rinaldi, email: [email protected]
Keywords: microbubbles; sonoporation; ultrasound; liver cancer; gene therapy
Received: June 05, 2018 Accepted: July 13, 2018 Published: August 14, 2018
ABSTRACT
Introduction: An innovative method, known as sonoporation, was used to induce the expression of silenced genes, such as (but not restricted to) TRAIL and p53, in liver cancer cells (HepG2). The principal aim of the present study was the re-activation of silenced apoptotic pathways in liver cancer models, by using diagnostic synovial microbubble as plasmid gene delivery tools in combination with epigenetic treatments.
Material and methods: HepG2 cells were used as a liver cancer model. Microbubbles (Sonovue®) were chosen as gene deliver system in combination with the sonoporation approach. Plasmid pEGFP-TRAIL and pEGFP-p53 were selected and propagated in Escherichia coli grown in LB broth, in order to obtain the necessary amount.
Results: Sonoporation was induced by using transducer (Sonitron 2000) and, among the several conditions tested, 3 MHz, 51% Duty Cycle, and 5 W/cm2, 30 s resulted as the best parameters. Data collected showed a dose dependent effect in terms of output energy. A transfection efficacy of 30 – 50% was achieved and recombinant gene expression induced apoptotic effects. In order to increase efficacy, we used the histone deacetylase inhibitor (HDACi, entinostat) MS-275, able to activate TRAIL and thus inducing a stronger pro-apoptotic effect in combination with TRAIL-gene re-expression.
Conclusion: For the first time, it was shown the possibility to induce the exogenous expression of the pro-apoptotic gene TRAIL and p53 in a liver cancer HepG2 cells via a sonoporation procedure. The epigenetic treatment using HDACi was able to increase the pro-apoptotic effects of the gene therapy.
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