Expression of coagulation factor IX (Christmas factor) in human hepatoma (HepG2) cell cultures after retroviral vector-mediated transfer.
Expression of coagulation factor IX (Christmas factor) in human hepatoma (HepG2) cell cultures after retroviral vector-mediated transfer.
复制标题
逆转录病毒载体介导的转移后人肝癌 (HepG2) 细胞培养物中凝血因子 IX（圣诞因子）的表达。
DOI：
10.1097/00043426-199305000-00007
复制
发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Kohn,DB
中科院分区：
文献类型：
--
作者：
Gordon,EM;Tang,H;Salazar,RL;Kohn,DB
目的在本研究中，我们比较了用巨细胞病毒（CMV）启动子控制的因子IX载体转导的HepG 2细胞的重组人因子IX的产生与非巨细胞病毒（CMV）启动子控制的因子IX载体转导的HepG 2细胞的内源性因子IX的产生。方法和结果Northern分析显示在原代肝细胞制备物中对应于已知大小的因子IX mRNA的2.8kb转录物和在原代肝细胞制备物中预期大小的载体因子IX转录物。转导的HepG 2细胞制备物。由转导的HepG 2细胞产生的因子IX被抗人因子IX的单特异性抗体完全抑制。Western分析表明，重组因子IX迁移到56 Kd的天然血浆因子IX的区域。由转导的HepG 2细胞的生物活性因子IX的生产是20倍大于由nontransduced原代hepatocyte.ConclusionThese数据表明，转导的HepG 2细胞转录，合成，并分泌正宗的因子IX，这些基因工程细胞分泌的因子IX的显着更大量的比非transduced原代肝细胞。研究正在进行中，以确定在转导的HepG 2细胞和原代肝细胞中肝细胞有丝分裂原对因子IX产生的影响。
PurposeIn this study, we compared production of recombinant human factor IX by HepG 2 cells transduced with a cytomegalovirus (CMV) promoter-controlled factor IX vector to endogenous production of factor IX by non-transduced primary rat hepatocytes.Methods and ResultsNorthern analysis showed 2.8 kb transcripts corresponding to the known size of factor IX mRNA in primary hepatocyte preparations and vector factor IX transcripts of the expected sizes in transduced HepG 2 cell preparations. Factor IX produced by transduced HepG 2 cells was completely inhibited by a mono-specific antibody against human factor IX. Western analysis showed that recombinant factor IX migrated to the region of native plasma factor IX at 56 Kd. Production of biologically active factor IX by transduced HepG 2 cells was 20-fold greater than that by nontransduced primary hepatocytes.ConclusionThese data indicate that transduced HepG 2 cells transcribe, synthesize, and secrete authentic factor IX, and that these genetically engineered cells secrete significantly greater amounts of factor IX than do non-transduced primary hepatocytes. Studies are in progress to determine the effect of hepatocyte mitogens on production of factor IX in transduced HepG 2 cells and primary hepatocytes.