ABSTRACT
This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellular 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 pshuttle-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 G(2)/M phase and apoptotic rate was increased, and the percentage of cells at G(0)/G(1) 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 single-gene-radiotherapy.
ABSTRACT
AIM: Through studying the differences between wild-type acidic fibroblast growth factor (waFGF) with recombinant aFGF (raFGF), the biological effect of raFGF concerned with mitogenic activity was evaluated. METHODS: NIH3T3 cell line was used. Cell proliferation with MTT method was used to study the mitogenic activity. Flow cytometry was also used for detection of apoptosis, cell membrane permeability and mitochondria potential. The role of heparin sulfate (HS) on aFGF biological effect was studied at the same time in this research. RESULTS: The enhancement of raFGF on cell proliferation was significantly lower than that of waFGF. The restriction of raFGF on apoptosis and the enhancement of it on cell membrane permeability were all lower than those of waFGF significantly. The enhancement of raFGF on mitochondria potential was lower than that of waFGF significantly. The HS improved the biological effect of aFGF. CONCLUSION: The mitogenic activity of raFGF is lower than that of waFGF and raFGF has little effect on apoptosis.