Cochet O, Kenigsberg M, Delumeau I, Virone-Oddos A, Multon M C, Fridman W H, Schweighoffer F, Teillaud J L, Tocqué B
Laboratoire de Biotechnologie des Anticorps, Institut Curie, Paris, France.
Cancer Res. 1998 Mar 15;58(6):1170-6.
Mutated ras genes are found in a large number of human tumors and, therefore, constitute one of the primary targets for cancer treatment. Microinjection of the neutralizing anti-Ras monoclonal antibody Y13-259 was previously reported to induce transient phenotypic reversion of ras-transformed rodent fibroblasts in vitro. We have prepared a single-chain Fv fragment (scFv) derived from Y13-259, and here, we show that intracellular expression of the scFv led to the specific inhibition of the Ras signaling pathway in Xenopus laevis oocytes and NIH3T3 fibroblasts. Moreover, neutralizing Ras with the scFv specifically promoted apoptosis in vitro in human cancer cells but not in untransformed cells. As a step toward cancer gene therapy, we finally demonstrated that intratumor transduction of HCT116 colon carcinoma cells with the anti-Ras scFv using an adenoviral vector elicited sustained tumor regression in nude mice.
在大量人类肿瘤中发现了突变的ras基因，因此，它构成了癌症治疗的主要靶点之一。先前有报道称，显微注射中和性抗Ras单克隆抗体Y13 - 259可在体外诱导ras转化的啮齿动物成纤维细胞发生短暂的表型逆转。我们制备了源自Y13 - 259的单链Fv片段（scFv），在此我们表明，scFv在细胞内的表达导致非洲爪蟾卵母细胞和NIH3T3成纤维细胞中Ras信号通路的特异性抑制。此外，用scFv中和Ras可在体外特异性促进人癌细胞凋亡，但对未转化细胞无此作用。作为癌症基因治疗的一个步骤，我们最终证明，使用腺病毒载体将抗Ras scFv转导至HCT116结肠癌细胞瘤内可在裸鼠中引起肿瘤持续消退。
