环孢素A作用于人滋养层细胞差异表达的功能基因  

Functional Genes Differentially Expressed in Human Trophoblast Cells Before and After Treated With Cyclosporin A

作　　者：杜美蓉[1] 闫凤亭[1] 李大金[1] 严缘昌[2] 李逸平[2] 金莉萍[1] 王明雁[1] 朱影[1] 袁敏敏[1] 孟毅[1] 

机构地区：[1]复旦大学妇产科研究所生殖免疫研究室,上海200011 [2]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031

出　　处：《中国药学杂志》2006年第3期189-192,共4页Chinese Pharmaceutical Journal

基　　金：上海市基础研究重点项目(03JC14016);复旦大学"985工程"项目基金

摘　　要：目的探讨环孢素A(cyclosporin A,CsA)作用于人滋养层细胞差异表达的功能基因。方法应用抑制性消减杂交筛选1.0μmol.L-1CsA作用于JAR细胞系前后差异表达的功能基因,经RT-PCR及蛋白质印迹进一步验证CsA作用前后原代分离的人早孕期滋养层细胞及JAR细胞株titin表达的变化。结果CsA作用于人JAR细胞系前后出现6个具有差异表达的基因,其中1个为已知基因titin,2个为功能未知的mRNA,3个为位于16号染色体上的EST。RT-PCR显示,CsA作用于人滋养层细胞24 h后出现titinmRNA的表达,72 h达高峰,并呈现明显的剂量依赖性,1.0μmol.L-1CsA作用最明显。蛋白质印迹显示,CsA可诱导titin的表达。结论CsA可能通过诱导滋养层细胞titin的高表达,改变其生物学行为,从而有利于胚胎着床及早期发育。OBJECTIVE To explore the functional gene expressions in human trophoblast cell induced by eyclosporin A （CsA）. METHODS The functional genes differentially expressed in JAR cell line treated with cyclosporin A were screened by means of suppression subtraetive hybridization （SSH）.The differential gene, titin, was testified by means of RT-PCR and Western blotting in first-trimester human trophoblast cells and JAR cell line. RESULTS There were six genes which expressed differentially in human trophoblast cells after being treated with cyclosporin A. Among the six differential expressed genes, one was known gene, titin, two were predicted genes, and three were ESTs located in chromosome 16. RT-PCR showed CsA induced titin mRNA expression in human trophoblast ceils after incubating with CsA for 24 h, and its expression was in a time dependent and dosage-dependent manner. Expression peak was reached after 72 h treatment with 1.0μmol· L^-1 CsA. Western blotting confirmed titin protein expression in human trophoblast cells. CONCLUSION CsA can induce titin high expression in human trophoblast cells and change its biological charactelistics so as to favor embryo implantation and early development.

关 键 词：环孢素A 滋养层细胞 抑制性消减杂交 反转录-聚合酶链反应 蛋白质印迹 肌肉巨球蛋白 

分 类 号：R979.5[医药卫生—药品]