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目的 探讨外源性血管抑素 K(1- 3)基因[angiostatin K(1- 3) ,AK(1- 3) ]在人脑胶质瘤中的表达及其作用 .方法 应用 PCR法 ,使人 Ig kappa链分泌信号肽序列S加装在 AK(1- 3)基因上 - SAK(1- 3) ;构建 SAK(1- 3)基因的真核表达载体 pc DNA- SAK(1- 3) .经脂质体法将其转染入SHG44人脑胶质瘤细胞 ,行 G418筛选获得转基因瘤细胞克隆 ,比较转基因和未转基因的瘤细胞生物学特性 .以免疫荧光、免疫组化和内皮细胞抑制实验检测瘤细胞表达的 AK(1-3)蛋白及其活性 .应用裸鼠皮下致瘤性实验 ,结合 SHG44瘤组织的各项检测 ,确定肿瘤表达 AK(1- 3)蛋白对人脑胶质瘤生长的影响 .结果 外源性 AK(1- 3)的表达不影响 SHG44瘤细胞的生物学特性 ,它能在体外抑制内皮细胞生长 ;表达AK(1- 3)的胶质瘤细胞体内致瘤性显著下降 ,抑瘤率为 76 .8% .结论 肿瘤表达的 AK(1- 3)蛋白能抑制其自身生长 ,该机制是通过抑制胶质瘤的血管生成 ,进而导致肿瘤局部缺血坏死实现的
Objective To investigate the expression of exogenous angiostatin K (1- 3) gene and its role in human glioma.Methods PCR method was used to detect the expression of angiostatin K (1- 3), AK (1- 3) SAK (1- 3) was inserted into the AK (1-3) gene of human Ig kappa chain and the eukaryotic expression vector pcDNA-SAK (1- 3) was constructed to construct SAK (1- 3) The recombinant plasmid was transfected into SHG44 human glioma cells by lipofectamine 2000. The clones of transgenic tumor cells were screened by G418 to compare the biological characteristics of transgenic and untransformed tumor cells.Furthermore, immunofluorescence, immunohistochemistry and endothelial cell The inhibitory activity of AK (1-3) protein and its activity in tumor cells was tested by using subcutaneous tumorigenicity test in nude mice and the detection of SHG44 tumor tissues to determine the effect of AK (1-3) The effect of exogenous AK (1- 3) expression on the proliferation and proliferation of SHG44 tumor cells was observed in vitro.Results The expression of exogenous AK (1- 3) did not affect the biological characteristics of SHG44 tumor cells and inhibited the growth of endothelial cells in vitro. The expression of AK (1- 3) The tumorigenicity was significantly decreased and the tumor inhibition rate was 76.8% .Conclusion AK (1- 3) protein expressed by tumor can inhibit its own growth by inhibiting Stromal tumor angiogenesis, leading to ischemia or necrosis of the tumor