质粒介导的组织激肽释放酶结合蛋白在肺癌相关细胞内的表达及其生物学活性研究  

Plasmid-mediated expression of kallistatin and its biological activity in lung cancer related cells

作　　者：汪宁卿 邹金[1] 刁勇[1] 

机构地区：[1]华侨大学生物医学学院,华侨大学分子药物研究院,福建泉州362021

出　　处：《药学学报》2013年第3期359-365,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(30973591,81271691);国际科技合作项目(2011DFG33320)

摘　　要：研究裸质粒能否转染肺癌相关细胞,且有效表达具有血管形成抑制活性的组织激肽释放酶结合蛋白(Kal),以及Kal的表达对体内外肺癌及相关细胞的生物学影响。采用Lipofectamine 2000介导质粒对肺癌相关细胞的转染,ELISA法检测Kal浓度,并考察Kal对细胞增殖、凋亡和迁移的影响。瘤内直接注射质粒后,观察NCI-H446皮下移植瘤内CD34、Ki-67和E-钙黏蛋白水平变化,以及肿瘤细胞凋亡的情况。转染质粒后,所有受试细胞均可表达Kal,其中HUVEC的增殖和迁移受到抑制;3种肺癌细胞NCI-H446、NCI-H460和A549的增殖被不同程度地抑制,并出现凋亡现象。体内研究显示,Kal可以抑制新生血管生成及肿瘤细胞增殖,肿瘤生长速度显著降低。因此,可介导Kal表达的裸质粒可以作为肺癌治疗的候选药物。This study is to investigate whether naked plasmid DNA can effectively transfect lung cancer related cells and express human kallistatin, an endogenous protein that inhibits angiogenesis and tumor growth, and to explore the biological activity of the low-level expressed kallistatin to lung cancer in vitro and in vivo. The plasmids were delivered with Lipofectamine 2000 to transfect various lung cancer related cells. Kal expression was determined by ELISA. The biological effects of Kal expression on proliferation, migration and apoptosis rate of the cells were examined. In subcutaneous NCI-H446 xenograft model, pKal was injected directly into tumors, the changes of CD34, Ki-67 and E-cadherin expression were detected with immunohistochemical assay, the tumor apoptosis was analyzed with TUNEL assay. Both the endothelial cell and lung cancer cells could express kallistatin after plasmid transfection. The proliferation and migration of human umbilical vein endothelial cells were inhibited, but the apoptosis rate was not affected. The proliferation rates of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited, and their apoptosis rates were enhanced, but different cells behaved differently. In subcutaneous NCI-H446 xenograft model, intratumor injection of pKal inhibited the growth of lung cancer by reducing angiogenesis and proliferation of tumor cells. In conclusion, this study demonstrated the efficacy of plasmid-mediated expression of kallistatin to lung cancer related cells, thus providing a basis for their clinical application in the treatment of lung cancer.

关 键 词：基因治疗 载体 组织激肽释放酶结合蛋白 新生血管形成 肺癌 

分 类 号：R963[医药卫生—微生物与生化药学]