Effects of bicistronic lentiviral vector-mediated herpes simplex virus thymidine kinase/ganciclovir system on human lens epithelial cells

	Effects of bicistronic lentiviral vector-mediated herpes simplex virus thymidine kinase/ganciclovir system on human lens epithelial cells
	Yang, J; Liu, TA; Lu, Y
刊名	CURRENT EYE RESEARCH
	2007
卷号	32 期号:1 页码:33-42
关键词	enhanced green fluorescent protein (EGFP) ganciclovir (GCV) herpes simplex virus-thymidine kinase (HSV-tk) human lens epithelial cell (HLEC) lentivirus suicide gene
通讯作者	Lu, Y (reprint author), Fudan Univ, Dept Ophthalmol, Eye & ENT Hosp, 83 Fenyang Rd, Shanghai 200031, Peoples R China.,luyi-eent@yahoo.com.cn
英文摘要	Posterior capsule opacification (PCO) is the most common complication after phacoemulsification cataract surgery. Hyperplasia of the lens epithelial cell after phacoemulsification is thought to be an important feature contributing to PCO. In this study, we investigated the feasibility of killing the human lens epithelial cells (HLECs) by lentivirus-mediated herpes simplex virus thymidine kinase (HSV-tk) gene/ganciclovir (GCV) in HLECs and studied the bystander effect. HLECs were infected with lentiviral vectors coexpressing HSV-tk and enhanced green fluorescent protein (EGFP) or expressing EGFP alone and treated with ganciclovir. Infection efficiency was assessed by fluorescence microscopy, fluorescence-activated cell sorting, and reverse transcription PCR. The cytotoxicity of the HSV-tk/GCV suicide gene therapy system was assessed by DNA ladder and electron microscopy. The time effect and bystander effect of HLEC growth inhibition were evaluated with cell proliferation assay. Lentiviral vector-mediated stable integration and efficient expression of HSV-tk in HLECs, with infection efficiency exceeding 95% GCV at concentrations of 15 similar to 25 mu g/ml, significantly induced apoptosis or necrosis of infected HLECs. GCV also killed normal cells mixed with HSV-tk infected cells. The bystander effect markedly increased the cytotoxicity of the HSV-tk/GCV system. Our results suggest that bicistronic lentiviral vectors can efficiently integrate several genes into HLECs and may be a gene therapy platform. Lentivirus-mediated suicide gene therapy might be a feasible treatment strategy to prevent capsule opacification.
学科主题	Ophthalmology
类目[WOS]	Ophthalmology
关键词[WOS]	POSTERIOR CAPSULE OPACIFICATION ; JUNCTIONAL INTERCELLULAR COMMUNICATION ; GENE-THERAPY ; SUICIDE GENE ; IN-VITRO ; TRANSDUCTION ; VIVO
收录类别	SCI
语种	英语
WOS记录号	WOS:000244142900005
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1514]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Yang, J,Liu, TA,Lu, Y. Effects of bicistronic lentiviral vector-mediated herpes simplex virus thymidine kinase/ganciclovir system on human lens epithelial cells[J]. CURRENT EYE RESEARCH,2007,32(1):33-42.
APA	Yang, J,Liu, TA,&Lu, Y.(2007).Effects of bicistronic lentiviral vector-mediated herpes simplex virus thymidine kinase/ganciclovir system on human lens epithelial cells.CURRENT EYE RESEARCH,32(1),33-42.
MLA	Yang, J,et al."Effects of bicistronic lentiviral vector-mediated herpes simplex virus thymidine kinase/ganciclovir system on human lens epithelial cells".CURRENT EYE RESEARCH 32.1(2007):33-42.