Enhanced Effects of TRAIL-endostatin-based Double-gene-radiotherapy on Suppressing Growth, Promoting Apoptosis and Inducing Cell Cycle Arrest in Vascular Endothelial Cells

• Published in: Journal of Huazhong University of Science and Technology. Medical sciences Vol. 32; no. 2; pp. 167 - 172
• Main Author: 李艳博 郭彩霞 王志成 龚平生 孙志伟 龚守良
• Format: Journal Article
• Language: English
• Published: Heidelberg Huazhong University of Science and Technology 01.04.2012; Key Laboratory of Radiobiology,Ministry of Health,School of Public Health,Jilin University,Changchun 130021,China%School of Public Health and Family Medicine,Capital Medical University,Beijing 100069,China%Key Laboratory of Radiobiology,Ministry of Health,School of Public Health,Jilin University,Changchun 130021,China%Key Laboratory for Molecular Enzymology and Enginnering of Ministry of Education,Jilin University,Changchun 130012,China; School of Public Health and Family Medicine,Capital Medical University,Beijing 100069,China
• Subjects: Annexin V; Apoptosis; Apoptosis - physiology; Apoptosis - radiation effects; Cell cycle; Cell Cycle Checkpoints - physiology; Cell Line; Cell Proliferation - radiation effects; Combined Modality Therapy - methods; Dose-Response Relationship, Radiation; Egr; endostatin; Endostatins - genetics; Endostatins - metabolism; Endothelial cells; Endothelial Cells - cytology; Endothelial Cells - physiology; Enzyme-linked immunosorbent assay; Flow cytometry; G1 phase; gene-radiotherapy; Genetic Therapy - methods; Humans; Ionizing radiation; Medicine; Medicine & Public Health; Plasmids; promoter; propidium iodide; Radiotherapy; Radiotherapy - methods; TNF-Related Apoptosis-Inducing Ligand - genetics; TNF-Related Apoptosis-Inducing Ligand - metabolism; TRAIL; TRAIL protein; Transfection; gene-radiotherapy; TRAIL; Egr-1 promoter; endostatin
• ISSN: 1672-0733; 1993-1352
• DOI: 10.1007/s11596-012-0030-x

This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellu-lar growth, apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro. The expression of TRAIL and endostatin protein in ECV304 cells was detected by ELISA after the transfection of recombinant plasmid pshuttle-Egr1-shTRAIL-shES and X-ray irradiation. Then MTT assay was used for determining the cellular proliferation, and flow cytometry (FCM) plus Annexin V and propidium iodide (PI) double-staining or PI single-staining were employed for the detection of apoptosis and cell cycle progression. The results showed that expression of TRAIL and endostatin protein exhibited a time- and dose-dependent change in ECV304 cells after pshut-tle-Egr1-shTRAIL-shES transfection in conjunction with irradiation. In the TRAIL-endostatin-based single- or double-gene-radiotherapy, the cell viability declined in a time- and dose-dependent manner, the percentage of cells at G2/M phase and apoptotic rate was increased, and the percentage of cells at G0/G1 phase was lowered as compared with those receiving radiotherapy alone. Moreover, TRAIL-endostatin-based double-gene-radiotherapy demonstrated better effects on growth inhibition, promotion of apoptosis and induction of cell cycle arrest in ECV304 cells than sin-gle-gene-radiotherapy.