Antiproliferative effect of double suicide gene delivery mediated by polyamidoamine dendrimers in human Tenon's capsule fibroblasts

Yang,Jin; Shi,Liu Kun; Sun,Hui Min; Wang,Yan Ming

doi:10.3892/etm.2017.5235

Antiproliferative effect of double suicide gene delivery mediated by polyamidoamine dendrimers in human Tenon's capsule fibroblasts

Authors:
- Jin Yang
- Liu Kun Shi
- Hui Min Sun
- Yan Ming Wang
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Affiliations: Tianjin Eye Hospital, Tianjin Key Laboratory of Ophthalmology and Vision Science, Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300020, P.R. China, Department of Ophthalmology, Tianjin Medical University, Tianjin 300384, P.R. China, Department of Glaucoma, College of Optometry and Ophthalmology, Tianjin Medical University Eye Institute, Tianjin Medical University Eye Hospital, Tianjin Medical University, Tianjin 300384, P.R. China, College of Pharmacy, Nankai University, Tianjin 300071, P.R. China
Published online on: September 29, 2017 https://doi.org/10.3892/etm.2017.5235
Pages: 5473-5479
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the therapeutic potential of a double suicide gene, thymidine kinase (TK) combined with cytosine deaminase (CD), mediated by generation of 5‑polyamidoamine dendrimers (G5‑PAMAM‑D) on human Tenon's capsule fibroblasts (HTFs) as an anti‑scarring agent. The pAcGFP1‑Hyg‑TK‑CD plasmid was transfected into HTFs, and reverse‑transcription polymerase chain reaction (RT‑PCR) was used to detect TK‑CD expression. MTT cell proliferation assay was used to evaluate the cytotoxic effects of ganciclovir (GCV) and 5‑flurocytosine (5‑FC) on HTFs. The optimal concentration of GCV and 5‑FC in TK‑CD transfected HTFs (HTF‑TK‑CD) was selected by accessing the lowest and highest cytotoxicity caused, respectively. The morphological changes of transfected HTFs following treatment with GCV and 5‑FC were observed by light and transmission electron microscopy. Results demonstrated that the double suicide gene TK‑CD mediated by the G5‑PAMAM‑D delivery system was successfully expressed in HTFs as determined by RT‑PCR. A concentration of 3 µg/ml GCV and 200 µg/ml 5‑FC was identified as optimal for these prodrugs. The growth rate and number of HTF‑TK‑CD cells decreased following treatment with GCV and 5‑FC as revealed by light microscopy. Additionally, the prodrugs GCV and 5‑FC not only demonstrated toxicity on transfected HTFs but also exerted a ‘bystander effect’. The present study illustrated that the double suicide gene TK‑CD delivery mediated by G5‑PAMAM‑D was effective in reducing HTF proliferation and inducing cell apoptosis. Furthermore, TK‑CD delivery mediated by G5‑PAMAM‑D may be used as an anti‑scarring agent and provide a therapeutic potential for patients requiring glaucoma filtration surgery.

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