Zeng L, Quilliet X, Chevallier-Lagente O, Eveno E, Sarasin A, Mezzina M
Laboratoire de Génétique Moléculaire, UPR 42 CNRS, Villejuif, France.
Gene Ther. 1997 Oct;4(10):1077-84. doi: 10.1038/sj.gt.3300495.
With the aim to devise a long-term gene therapy protocol for skin cancers in individuals affected by the inherited autosomal recessive xeroderma pigmentosum we transferred the human DNA repair XPA, XPB/ERCC3 and XPC cDNAs, by using the recombinant retroviral vector LXSN, into primary and immortalized fibroblasts obtained from two XP-A, one XP-B (associated with Cockayne's syndrome) and two XP-C patients. After transduction, the complete correction of DNA repair deficiency and functional expression of the transgenes were monitored by UV survival, unscheduled DNA synthesis and recovery of RNA synthesis, and Western blots. The results show that the recombinant retroviruses are highly efficient vectors to transfer and stably express the human DNA repair genes in XP cells and correct the defect of DNA repair of group A, B and C. With our previous results with XPD/ERCC2, the present work extends further promising issues for the gene therapy strategy for most patients suffering from this cancer-prone syndrome.
为了设计一种针对遗传性常染色体隐性色素性干皮病患者皮肤癌的长期基因治疗方案，我们使用重组逆转录病毒载体LXSN，将人类DNA修复基因XPA、XPB/ERCC3和XPC的cDNA，导入从两名XP-A患者、一名XP-B患者（与科凯恩综合征相关）和两名XP-C患者获得的原代及永生化成纤维细胞中。转导后，通过紫外线存活率、非定标DNA合成、RNA合成恢复以及蛋白质免疫印迹法，监测DNA修复缺陷的完全纠正和转基因的功能性表达。结果表明，重组逆转录病毒是在XP细胞中转移并稳定表达人类DNA修复基因、纠正A、B和C组DNA修复缺陷的高效载体。结合我们之前关于XPD/ERCC2的研究结果，目前的工作为大多数患有这种易患癌症综合征的患者的基因治疗策略进一步拓展了有前景的方向。
