重组pegr-endostatin-egfp质粒的辐射诱导特性及其在基因-放射治疗中的应用-radiation - induced characteristics of recombinant pegr - endostatin - egfp plasmid and its application in gene - radiotherapy.docxVIP

汕头大学医学院博士学位论文 汕头大学医学院 博士学位论文 IV IV expression property of endostatin was investigated by RT-PCR and that of EGFP was detected by flow cytometry. Tumor-bearing mice were treated by the plasmid injection and 2 Gy X-irradiation for 3 times. Tumor growth was observed for 18 d after treatment. Change of tumor capillary formation was measured with histochemistry assay at the end of the experiment. [RESULTS]: It was confirmed that pEgr-Endostatin-EGFP recombinant plasmid had been constructed successfully by identification of Restriction Endonuclease Cleavage Reactions and PCR. After RT-PCR, there was no special band of endos tatin in the normal B16 cells, but the positive bands were observed in pEgr-Endostatin- EGFP transfected groups with or without irradiation.The expression of GFP in B16 melanoma cells was detected after X- irradiation with 0.05~20 Gy. Time-course studies showed that the expression of GFP in B16 cells reached its peak at 8 h after 2 Gy X-ray irradiation. The injection of pEgr-Endostatin- EGFP recombinant plasmid into the implanted B16 melanoma in C57BL/6J mice followed by local X- irradiation could significantly inhibit tumor growth with inhibition of intratumor micro-vessel density. [CONCLUSIONS] ： pEgr-Endostatin- EGFP recombind plasmid had been constructed successfully. Endostatin genes combined with irradiation are able to suppress the growth of B16 melanoma cells significantly in vitro and its inhibition effects were better than that in gene- or irradiation- treatment alone. The experimental results in tumor-bearing mouse model showed that gene-radiotherapy could significantly inhibit tumor growth with inhibition of intratumor micro- vessel density. The results are of substantial importance in investigating a new approach for clinical treatment of tumors as well as in improving radiotheraputic effects. KEY WORDS: gene-radiotherapy; radiotherapy; gene therapy; Endostatin; melanoma PAGE PAGE VI 英文缩略词表 缩略语 VEGF 英文全名 vascular endothelial g