Tumoricidal effect of human olfactory ensheathing cell mediated suicide gene therapy in human glioblastoma cells

• Published in: Molecular biology reports Vol. 45; no. 6; pp. 2263 - 2273
• Main Authors: Hashemi, Mansoureh, Hadjighassem, Mahmoudreza, Zali, Alireza, Hashemi, Javad
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.12.2018; Springer Nature B.V
• Subjects: Adult; Animal Anatomy; Animal Biochemistry; Antiviral Agents; Apoptosis; Astrocytes; Astrocytes - drug effects; BAX protein; Bcl-2 protein; Biomedical and Life Sciences; Biopsy; Brain cancer; Bystander Effect; Cell culture; Cell death; Chemical communication; Coculture Techniques; Connexin 43; Connexin 43 - drug effects; DNA fragmentation; Fluorescent indicators; Ganciclovir; Ganciclovir - administration & dosage; Ganciclovir - pharmacology; Gap junctions; Gap Junctions - drug effects; Gene therapy; Genes, Reporter; Genetic Therapy; Genetic Vectors; Glioblastoma; Glioblastoma - drug therapy; Glioblastoma cells; Herpes simplex; Herpesvirus 1, Human; Histology; Humans; Life Sciences; Morphology; Mucosa; Olfactory ensheathing cells; Olfactory epithelium; Olfactory Mucosa; Original Article; Primary Cell Culture; Suicide genes; Thymidine; Thymidine kinase; Thymidine Kinase - therapeutic use; Tumor Cells, Cultured; Suicide gene therapy; Bystander effect; Olfactory ensheathing cells; Glioblastoma multiforme; Apoptosis
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-018-4388-0

The potential of herpes simplex virus type 1 thymidine kinase (HSV-tk)-expressing olfactory ensheathing cells (OEC) treated with ganciclovir (GCV) to induce cell death in adjacent HSV-tk-negative cells (bystander effect) has been well demonstrated. Although it has been shown that bystander effect occurs through the delivery of phosphorylated GCV, the bystander effect mechanism and the role of gap junctions for human OECs mediated suicide gene therapy in primary astrocytes of human glioblastma remain obscure. In the present study, the efficacy of a new method for the transfer of phosphorylated GCV from OECs into primary astrocytes was evaluated. Surgical biopsy of glioblastoma was used to isolate primary astrocyte. Biopsy of olfactory mucosa was applied to isolate olfactory ensheathing cell. Expression of S100-beta antigen was confirmed immunocytochemically in astrocytes and OECs. OECs were transduced to lentiviral containing thymidine kinase gene (TK) and co-cultured with astrocytes. Fluorescent dye transfer and western blot analysis indicated the expression of connexin43 between olfactory ensheathing cells and astrocytes whereas, expression of the gap junction protein connexin43 was inhibited by the gap junction inhibitor 18α-glycyrrhethinic acid (AGA, 20 µg/ml). Furthermore, co-culture of astrocytes with OEC-TK in the presence of concentration of 30 µg/ml GCV led to a decrease in astrocytes survival rate. Also, apoptosis hallmarks, including DNA fragmentation in cell nuclear, expression increase of Bax to Bcl-2 ratio and increase of caspase3 activation were observed in this study. Our findings suggest that human olfactory ensheathing cells can deliver phosphorylated GCV into the glioblastoma derived astrocytes through gap junction communication for apoptosis induction.