Intracellular expression of an antibody fragment-neutralizing p21 ras promotes tumor regression.
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.
Cochet
O
O
Laboratoire de Biotechnologie des Anticorps, Institut Curie, Paris, France.
Kenigsberg
M
M
Delumeau
I
I
Virone-Oddos
A
A
Multon
M C
MC
Fridman
W H
WH
Schweighoffer
F
F
Teillaud
J L
JL
Tocqué
B
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage; D017209:Apoptosis; D016158:Genes, p53; D011905:Genes, ras; D006801:Humans; D007128:Immunoglobulin Fragments / Q000008:administration & dosage*; D007167:Immunotherapy; D051379:Mice; D008819:Mice, Nude; D008845:Microinjections; D009374:Neoplasms, Experimental / Q000628:therapy*; D016283:Proto-Oncogene Proteins p21(ras) / Q000037:antagonists & inhibitors* / Q000276:immunology; D015398:Signal Transduction; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)