论文部分内容阅读
目的 :构建arrestencDNA的分泌型真核表达载体 ,将其转染人胃癌SGC 790 1细胞 ,探讨arresten体外抑制血管内皮细胞生长的活性。方法 :利用PCR法将小鼠Igκ链信号序列的 6 0个碱基拼接至arrestencDNA的氨基端 ,并克隆入真核表达载体pcDNA3.1(+) ,然后以脂质体法将重组pcDNA3.1(+) arresten转染SGC 790 1细胞 ,Western印迹检测arresten的分泌表达 ,最后 ,提取SGC 790 1细胞培养上清 ,采用内皮细胞增殖抑制试验鉴定上清中arresten活性。结果 :成功构建了arrestencDNA的分泌型真核表达载体 ,获得可表达arresten的SGC 790 1细胞系 ,真核表达的arresten具有抑制血管内皮细胞增殖的活性。结论 :arresten是一种有效的血管生成抑制剂 ,其cDNA的真核表达载体的成功构建为针对人类胃癌的arresten抗血管基因治疗研究奠定基础。
OBJECTIVE: To construct the secreting eukaryotic expression vector of arresten cDNA and transfect it into SGC-7901 human gastric cancer cell line SGC-7901 to investigate the inhibitory effect of arresten on the growth of vascular endothelial cells in vitro. Methods: Sixty base pairs of mouse Ig kappa chain signal sequence were spliced to the amino terminal of arresten cDNA by PCR and cloned into the eukaryotic expression vector pcDNA3.1 (+). The recombinant pcDNA3.1 (+) arresten transfected SGC 790 1 cells, Western blot detection of arresten secretion, and finally extracted SGC 790 1 cell culture supernatant, inhibition of endothelial cell proliferation was identified arresten activity in the supernatant. Results: The secreting eukaryotic expression vector of arresten cDNA was successfully constructed and the arresten-expressing SGC 7901 cell line was obtained. The eukaryotic expression of arresten inhibited the proliferation of vascular endothelial cells. Conclusion: Arresten is a potent angiogenesis inhibitor. The successful construction of its eukaryotic expression vector for cDNA laid the foundation for arresten anti-vascular gene therapy in human gastric cancer.