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休眠蛋白 (restin)是内皮抑素 (endostatin)的同源蛋白 ,它们之间相同的氨基酸占整个蛋白质的 6 2 %。休眠蛋白位于胶原XV羧端的非胶原结构域。在大肠杆菌中重组表达的休眠蛋白经复性后可以专一地抑制牛主动脉内皮细胞的生长 ,并引起凋亡。将鼠源的休眠蛋白基因与信号肽连接后克隆入真核表达载体 pCDNA3,取名为pCDNAXV并转染Bel74 0 4细胞。利用RT PCR和Western印迹证实休眠蛋白在稳定转染株中表达。将这一稳定转染株细胞接入裸鼠皮下观察肿瘤生长 ,发现转染有休眠蛋白基因的肿瘤生长显著慢于对照组 ;由于休眠蛋白基因对肿瘤细胞的增殖没有影响 ,这种抑制作用很可能是通过抑制血管生长来实现的。同时 ,这种方法为研究抑制血管生长蛋白提供了新的手段
Restin is a homologous protein of endostatin, with the same amino acids accounting for 62% of the total protein. The dormant protein is located in the non-collagenous domain at the carboxy-terminus of collagen XV. The recombinant protein expressed in Escherichia coli can restrain the growth of bovine aortic endothelial cells and cause apoptosis specifically after refolding. The mouse dormant protein gene was ligated with the signal peptide and cloned into the eukaryotic expression vector pCDNA3, which was named pCDNAXV and transfected into Bel74 0 4 cells. The expression of dormant protein in stable transfectants was confirmed by RT PCR and Western blotting. The stable transfected cells were injected into the nude mice subcutaneously to observe the tumor growth and found that the transfection of the dormant protein gene of tumor growth was significantly slower than the control group; as the dormant protein gene has no effect on the proliferation of tumor cells, this inhibition is May be achieved by inhibiting blood vessel growth. At the same time, this method provides a new means of studying the protein that inhibits the growth of blood vessels