Kobayashi N, Noguchi H, Westerman K A, Matsumura T, Watanabe T, Totsugawa T, Fujiwara T, Leboulch P, Tanaka N
First Department of Surgery, Okayama University Medical School, Japan.
Cell Transplant. 2000 Sep-Oct;9(5):737-42. doi: 10.1177/096368970000900525.
A worldwide shortage of donor livers is a limiting factor of the clinical application of hepatocyte transplantation (HTX). To resolve this issue, we focused on a reversible immortalization system that allows temporary expansion of primary hepatocyte populations by transfer of an oncogene that can be subsequently excised. As a preliminary test toward this goal, we examined the efficacy of Cre/loxP site-specific recombination in a transformed human liver cell line, HepG2. The present study utilized retroviral transfer of a prototypical immortalizing gene, simian virus 40 large T antigen (SV40Tag), flanked by a pair of loxP recombination targets and adenovirus-mediated Cre/loxP recombination. Here we report that complete elimination of the retroviral transferred oncogene was achieved by site-specific recombination using a replication-deficient recombinant adenovirus vector producing Cre recombinase (Ad-Cre).
供体肝脏的全球短缺是肝细胞移植（HTX）临床应用的一个限制因素。为了解决这个问题，我们专注于一种可逆的永生化系统，该系统通过转移随后可切除的癌基因来实现原代肝细胞群体的临时扩增。作为朝着这个目标的初步测试，我们检测了Cre/loxP位点特异性重组在转化的人肝癌细胞系HepG2中的效果。本研究利用了由一对loxP重组靶点侧翼的原型永生化基因猿猴病毒40大T抗原（SV40Tag）的逆转录病毒转移以及腺病毒介导的Cre/loxP重组。在此我们报告，使用产生Cre重组酶的复制缺陷型重组腺病毒载体（Ad-Cre）通过位点特异性重组实现了逆转录病毒转移的癌基因的完全消除。
