血管内皮生长因子-C反义多肽对人骨肉瘤细胞生物学行为影响的研究

目的检测血管内皮生长因子-C（VEGF—C）反义多肽对人骨肉瘤细胞的生物学活性的影响，探讨VEGF—C反义多肽用于骨肉瘤基因治疗的可行性。方法制备靶向VEGF—C的反义多肽，通过与骨肉瘤细胞株共培养，噻唑蓝（MTT）法检测细胞增殖抑制率，人工基底膜侵袭实验检测细胞侵袭能力；构建骨肉瘤细胞裸鼠皮下移植瘤模型，瘤体内注射VEGF—C反义多肽，观察其对骨肉瘤细胞的抑制作用；Western Blot检测VEGF—C在瘤组织的表达情况；免疫组织化学分析肿瘤组织VEGFR-3阳性脉管密度。结果VEGF—C反义多肽具有较强的生物学活性，对骨肉瘤细胞具有明显的增殖抑制作用，其抑增殖效应呈时间和浓度依赖性；瘤组织内给予VEGF—C反义多肽后，Western blot分析VEGF—C在瘤组织中的表达降低，肿瘤生长受制，抑瘤率为52％（P〈0．05）；免疫组化分析VEGF—C反义多肽可抑制肿瘤组织VEGFR-3阳性脉管形成。结论VEGF—C具有良好的生物学活性，可通过抑制骨肉瘤细胞的增殖、侵袭转移能力及肿瘤组织VEGFR-3阳性脉管形成而发挥抗骨肉瘤作用。

Studying on the cellular Biologic behavior effects of human antisense-VEGF gene C antisense polypeptide to human osteosarcoma

Objective To detect the cellular Biologic behavior effects of on human antisense - VEGF gene C antisense polypeptide to human osteosarcoma. Investigating the possibility of antisense - VEGF gene C for of genetic therapy human osteosarcoma. Methods Preparation of target to antisense -VEGF gene C antisense polypeptides, by it and the human osteosarcoma cell strain training, detecting the cell proliferation inhibiting rate by the methods of MTT, detecting the cellinvasive ability by manual basal membrane;constructing osteosarcoma cells invasive mole in nude mice,intratumor injection antisense -VEGF gene C antisense polypeptide, observeing the inhibition role to osteosarcoma cells, Western Blot Study on the expression status in tumor tissues ;analysising on tumor tissues antisense -VEGFR -3 positive vessel density by immunohistochemistry. Resuits The antisense - VEGF gene C has more strong biological activity, it has obvious proliferative inhibitory effect to osteosarcoma, its repress and proliferative effect showed time and concentration dependence;After giving antisense polypeptide in tumor tissue, Western blot analysis on antisense - VEGF gene Cexpression reduced in tumor tissue, the Tumor Growth was controlled, tumor inhibitory rate was 52% （ P 〈 0.05 ） ; immtmohistochemistry antisense - VEGF gene C antisense polypeptide can inhibit tumor tissues the formation of antisense - VEGFR - 3 positive vessel. Conclusion The antisense - VEGF gene C has strong biological activity,it can play the effect of resistance tumor tissues by controlling the proliferation and invasion abilities of tumor tissues and the formation of antisense - VEGFR - 3 positive vessel.