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The purpose of this study is to develop a specific and efficient targeted gene therapy candidate approach for laryngeal carcinomas. Several promoters of human squamous cell carcinoma antigen 2(SCCA2), secretory leukocyte protease inhibitor (SLPI) and Survivin genes were cloned from human genomic DNA and evaluated for tumor-specific transcription potential in human laryngeal carcinoma Hep-2 cells by dual luciferase assays. One SLPI promoter fragment (677 bp) showed the highest efficiency and specificity, and was used to control the expression of a recombinant active caspases-3 (revCasp3), which could trigger apoptosis without activation of its upstream cascade elements once expressed in a cell, in an adenoviral vector (Ad-SLPI-revCasp3), and its antitumor efficacy was assessed. In vitro infection with Ad-SLPI-revCasp3 showed revCasp3 could be specifically expressed in Hep-2 cells, resulting in efficient activation of endogenous Caspase-3 and subsequent apoptosis of Hep-2 cells. In Hep-2 nude mice xenograft model, intratumoral administration of Ad-SLPI-revCasp3 significantly inhibited tumor growth without obvious loss of body weight and obvious hepatic toxicity. In summary, our study showed the specific and efficient apoptosis-inducing potential of Ad-SLPI-revCasp3, and this makes it a new candidate approach of targeted gene therapy for laryngeal squamous cell carcinoma, which needs further systematic investigation.

Purpose To investigate the toxic effects of ethylenediaminetetraacetic acid disodium salt (EDTA), a corneal penetration enhancer in topical ophthalmic formulations, on DNA in human corneal epithelial cells (HCEs), and to investigate whether the effect induced by EDTA can be inhibited by high molecular weight hyaluronan (HA). Methods Cells were exposed to EDTA in concentrations ranging from 0.00001 to 0.01% for 60 min, or 30 min high molecular weight HA pretreatment followed by EDTA treatment. The cell viability was measured by the MTT test. Cell apoptosis was determined with annexin V staining by flow cytometry. The DNA single-and double-strand breaks of HCEs were examined by alkaline comet assay and by immunofluorescence microscope detection of the phosphorylated form of histone variant H2AX (gH2AX) foci, respectively. Reactive oxygen species (ROS) production was assessed by the fluorescent probe, 2 0 ,Results EDTA exhibited no adverse effect on cell viability and did not induce cell apoptosis in human corneal epithelial cells at concentrations lower than 0.01%. However, a significant increase of DNA single-and double-strand breaks was observed in a dose-dependent manner with all the concentrations of EDTA tested in HCEs. In addition, EDTA treatment led to elevated ROS generation. Moreover, 30 min preincubation with high molecular weight HA significantly decreased EDTA-induced ROS generation and DNA damage. Conclusions EDTA could induce DNA damage in HCEs, probably through oxidative stress. Furthermore, high molecular weight HA was an effective protective agent that had antioxidant properties and decreased DNA damage induced by EDTA.

Aims To evaluate the involvement of multipotential stem and progenitor cells in the pathogenesis of pterygium. Methods Paraffin-embedded and snap-frozen primary pterygium (n ¼ 10) were serially sectioned and analysed immunohistochemically to determine the expression level of AC133 (marker for the primitive haematopoietic progenitors), CD34 (marker for the haematopoietic progenitor cells and endothelium), c-Kit (marker for haematopoietic and stromal progenitor cells), and STRO-1 (a differentiation antigen present on bone marrow fibroblast cells and on various nonhaematopoietic progenitor cells). Results In all the primary pterygium, immunoreactivity of AC133 and STRO-1 was found in some of the epithelial and stromal cells, CD34 was observed in the vascular endothelium, and some scattered ovoidal cells were found in the subepithelial connective tissue. C-Kit was expressed mainly in the basal epithelium of the head portions, and some spindle-shaped stromal cells. There is no immunoreactivity of AC133, c-Kit, and STRO-1 in normal conjunctiva, whereas CD34 was mildly stained with vessel wall. Conclusion Multipotential stem and progenitor cells may be involved in the pathogenesis of pterygium through its differentiation into fibroblasts and vascular endothelial cells.