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作 者:周可祥[1] 吴秉铨[1] 郑杰[1] 王洁良[1] 由江峰[1] 崔湘琳[1]
机构地区:[1]北京医科大学病理学教研室,第一军医大学病理学教研室
出 处:《中华医学杂志》1994年第7期402-405,共4页National Medical Journal of China
基 金:国家"八五"攻关基金
摘 要:利用基因重组技术构建了一个含有全长人TIMP-1cDNA的真核表达重组体。重组质粒通过脂质体法导入高转移入肺巨细胞癌细胞系(PG细胞)。随机挑选5个G418抗性克隆,并对其中的两个克隆PG-T2和PGT4进行了深入的研究。Northern印迹表明,这两个克隆具有较高的TIMP-1RNA表达,其表达水平明显高于PG细胞和空载体转染对照细胞PG-MV。PG-T2和PG-T4克隆细胞显示出很高的TIMP-1蛋白活性,相同条件下PG和PG-MV未能显示出具有TIMP-1蛋白活性。此外,与其母系PG细胞及空载体转染对照细胞PG-MV相比,TIMP-1转染细胞的增殖速度和侵袭能力明显下降,其在软琼脂上形成克隆和在裸鼠体内成瘤的能力丧失。提示PG细胞中TIMP-1基因的特异性上调不仅能抑制其侵袭能力,而且对其增殖和成瘤产生抑制作用。recombinant plasmid, which contains a fulllenth cDNA of human tissue inhibitor ofmetalloproteinases-l (TIMP-l), was constructed byusing gene recombinant technique, and introduced in-to a highly metastatic human giant eell carcinoma(PG) by lipofectin technique. Two of the transfectants,PG-T2 and PG-T4, were studied thoroughly. Northern blot showed an increased level of TIMP-1mRNA in PG-T2 and PG-T4, compared with PGand PG-MV1 (vector-transfected control). The twotransfectants also exhibited higher TIMP-1 proteinactivities. Moreover, they showed significant reductionin their proliferation rate and invasive abilities. Theabilities of forming colonies in soft agar andtumorigenecity in athymic nude mice were abrogatedin PG-T2 and PG-T4. The preliminary results sug-gest that a specific upregulation of TIMP-l expres-sion in metastatic cells could not only supress theirinvasive and metastatic phenotype, but also inhibittheir proliferation and tumorigenecity.
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