Baculovirus-based gene silencing of Humanin for the treatment of pituitary tumors
- 221 Downloads
Pituitary tumors are the most common primary intracranial neoplasms. Humanin (HN) and Rattin (HNr), a rat homolog of HN, are short peptides with a cytoprotective action. In the present study, we aimed to evaluate whether endogenous HNr plays an antiapoptotic role in pituitary tumor cells. Thus, we used RNA interference based on short-hairpin RNA (shRNA) targeted to HNr (shHNr). A plasmid including the coding sequences for shHNr and dTomato fluorescent reporter gene was developed (pUC-shHNr). Transfection of somatolactotrope GH3 cells with pUC-shHNr increased apoptosis, suggesting that endogenous HNr plays a cytoprotective role in pituitary tumor cells. In order to evaluate the effect of blockade of endogenous HNr expression in vivo, we constructed a recombinant baculovirus (BV) encoding shHNr (BV-shHNr). In vitro, BV-shRNA was capable of transducing more than 80% of GH3 cells and decreased HNr mRNA. Also, BV-shHNr increased apoptosis in transduced GH3 cells. Intratumor injection of BV-shHNr to nude mice bearing s.c. GH3 tumors increased the number of apoptotic cells, delayed tumor growth and enhanced survival rate, suggesting that endogenous HNr may be involved in pituitary tumor progression. These preclinical data suggests that the silencing of HN expression could have a therapeutic impact on the treatment of pituitary tumors.
KeywordsHumanin Rattin Baculovirus ShRNA Pituitary tumor Apoptosis
This work was supported by the Consejo Nacional de Investigaciones Científicas y Tecnológicas, National Research Council (PIP 11420110100353 to M.C.; PIP 11220120100261 to A.S, doctoral fellowships to MFG, MP, ASA), Agencia Nacional de Promoción Científica y Tecnológica (PICT 2013-0310, PICT-2015-3309 to M.C.; PICT 2014-0334 to A.S.; PICT 2014-1827 to V.R., doctoral fellowship to CZ), the University of Buenos Aires (20020130100020 to A.S.) and the National University of La Plata (X703 to V.R.).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the Ethics Committee of the School of Medicine, University of Buenos Aires.
- 19.Pidre ML, Ferrelli ML, Haase S, Romanowski V (2013) Baculovirus display: a novel tool for vaccination. In: Romanowski V (ed) Biochemistry genetics and molecular biology. Intech, Rijeka, pp 137–164Google Scholar
- 29.Mottaghi-Dastjerdi N, Soltany-Rezaee-Rad M, Sepehrizadeh Z, Roshandel G, Ebrahimifard F, Setayesh N (2014) Genome expression analysis by suppression subtractive hybridization identified overexpression of Humanin, a target gene in gastric cancer chemoresistance. Daru 22:14CrossRefPubMedPubMedCentralGoogle Scholar