ATRA enhances the bystander effect of suicide gene therapy driven by the specific promoter LEP 503 in human lens epithelial cells
文献类型：期刊论文
| 作者 | Yang, J; Liu, TJ; Jiang, YX; Lu, Y |
| 刊名 | MOLECULAR VISION
 |
| 出版日期 | 2012 |
| 卷号 | 18期号:1页码:2053-2066 |
| 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 | Purpose: To establish a novel, targeted lentivirus-mediated LEP503-HSV-tk/GCV suicide gene therapy system combined with all trans-retinoic acid (ATRA) for the inhibition of human lens epithelial cell (HLEC) proliferation and treatment of posterior capsular opacification (PCO) after cataract surgery; to estimate the enhancement of the bystander effect by ATRA; and to explore the role of Connexin43 (Cx43) mediated gap junctional intercellular communication (GJIC) in the bystander effect of the HSV-K/GCV system. Methods: A Lenti-LEP503-HSV-tk-EGFP vector was generated by cloning the lens-specific promoter LEP503 (lens specific promoter 503) from genomic DNA of HLECs by PCR. The vector was then inserted into the promoter-less vector from lentivirus-based (CMV)-HSV-tk-EGFP. The expressional specificity of the LEP503 promoter was assessed by investigating the expression of EGFP (enhanced green fluorescent protein) and HSV-tk (herpes simplex virus thymidine kinase) mRNA, both driven by Lenti-LEP503-HSV-tk-EGFP vector, by fluorescence microscopy, RT-PCR, flow cytometry, and western blot assays in HLECs, human adult retinal pigment epithelium cells (RPECs), human adult skin fibroblast cells (ASFCs), and Hela cells. Morphological changes were observed by fluorescence microscopy and cell viability was determined using the Cell Counting kit-8 Cell Proliferation (CCK-8) and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays after Lenti-LEP503-HSV-tk/GCV system combined with ATRA treatment on HLECs. Flow cytometry, DNA fragmentation, and western blot assays were employed to analyze the mechanisms of bystander effects. Results: The promoter LEP503-mediated HSV-tk was specifically expressed in HLECs, and ATRA dose-dependently strengthened the bystander effect following LEP503-mediated HSV-tk/GCV gene therapy against lens cells by upregulating the expression of the gap junction protein Cx43. Conclusions: The Lenti-LEP503-HSV-tk/GCV suicide gene therapy system, combined with ATRA as an adjuvant, may be a feasible supplementary method for PCO treatment that targets residual lens cells. |
| 学科主题 | Biochemistry & Molecular Biology; Ophthalmology |
| 类目[WOS] | Biochemistry & Molecular Biology ; Ophthalmology |
| 关键词[WOS] | TRANS-RETINOIC ACID ; JUNCTIONAL INTERCELLULAR COMMUNICATION ; POSTERIOR CAPSULAR OPACIFICATION ; VERSUS-HOST-DISEASE ; THYMIDINE KINASE ; CANCER ; PREVENTION ; VIRUS ; DIFFERENTIATION ; INHIBITION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000307045800002 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/698]  |
| 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, TJ,Jiang, YX,et al. ATRA enhances the bystander effect of suicide gene therapy driven by the specific promoter LEP 503 in human lens epithelial cells[J]. MOLECULAR VISION,2012,18(1):2053-2066. |
| APA | Yang, J,Liu, TJ,Jiang, YX,&Lu, Y.(2012).ATRA enhances the bystander effect of suicide gene therapy driven by the specific promoter LEP 503 in human lens epithelial cells.MOLECULAR VISION,18(1),2053-2066. |
| MLA | Yang, J,et al."ATRA enhances the bystander effect of suicide gene therapy driven by the specific promoter LEP 503 in human lens epithelial cells".MOLECULAR VISION 18.1(2012):2053-2066. |
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