摘要
OSM是一种对黑色素瘤细胞显示抑制作用的细胞因子.为进行OSM针对黑色素瘤的基因-放射治疗研究,构建了小鼠Egr-1基因调控序列引导入OSMcDNA真核表达质粒(pEO),pEO质粒转染小鼠B-16黑色素瘤细胞,经G418和抗人OSM抗体的筛选,获得了稳定表达OSM的克隆细胞(pEO-1细胞),OSM表达量可达5.97ng每105细胞天,分子量为32kD.pEO-1细胞用一定浓度H2O2处理后OSM表达量可提高62%。
Oncostatin M(OSM),a single chain glycoprotein cytokine produced by stimulated macrophages and T lymphocytes,was previously identified by its ability to inhibit in vitro the proliferation of cells from melanoma and other solid tumors.To prove the potential application of OSM in the gene radiotheraphy for melanoma in vivo,a human OSM cDNA recombinant expression vector directed by Egr 1 gene promoter (pCl neo Egr 1R hOSM cDNA,pEO) was constructed and used to transfect mouse B 16 melanoma cells.The cloned cells secreting OSM continuously had been selected with G418 and anti hOSM antibodies,and the amount of secreted OSM in serum free supernatants was about 5 97 ng(per day 10 5 cells) determined by ELISA and with an apparent molecular weight of 32 kD by SDS electrophoresis and Western blot.It was also found that 62% of increased secretion of OSM from the pEO transfected cells was achieved as compared with control levels when H 2O 2 was added in medium,indicating that the Egr 1R hOSM construct could be activated by oxygen radicals so as to enhance the expression of OSM gene.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1999年第1期1-5,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金