A novel single‐chain variable fragment antibody against FGF‐1 inhibits the growth of breast carcinoma cells by blocking the intracrine pathway of FGF‐1

Abstract

SummaryThe fibroblast growth factors (FGFs) are important for embryo development, wound healing, hematopoiesis, and angiogenesis. FGF-1, a member of FGF family, is involved in both receptor-dependent pathways and an intracrine pathway. Studies have recently shown that FGF-1 is overexpressed in the early stages of several kinds of cancer. Thus, FGF-1 is a candidate for cancer immunotargeting. To study the potential use of therapeutic antibodies against FGF-1, a monoclonal hybridoma 1C9 secreting monoclonal antibody specific for FGF-1 was developed. Then, a single-chain variable fragment (scFv) antibody was genetically engineered from hybridama 1C9. The binding of the scFv1C9 to the antigen FGF-1 was demonstrated by ELISA and immunoprecipitation assays. Functional analysis showed that the overexpressed scFv1C9 in MCF-7 cells targeted endogenous FGF-1 and prevented the translocation of FGF-1 into the nucleus, resulting in the blockade of the intracrine pathway of FGF-1, which caused the G1 arrest by p21 up-regulation. These results suggest that the generated scFv1C9 is an effective inhibitor of the intracrine pathway of FGF-1 and has a potential application as anti-tumoral agent in breast cancer.

A simple strategy to obtain ultra-sensitive single-chain fragment variable antibodies for aflatoxin detection

“…Mouse immunoglobulin VH and VL genes were amplied for each mAb using the primers in the RPAS Mouse scFv Module (GE Healthcare Bio-Sciences, Shanghai, China). 16 The following thermal conditions were used for polymerase chain reaction (PCR) analysis of the VH genes: 30 cycles of 94 C for 30 s, 65 C for 30 s, and 72 C for 40 s, followed by a nal extension at 72 C for 7 min; the following thermal conditions were used for PCR analysis of the VL genes: 25 cycles of 94 C for 30 s, 65 C for 1 min, and 72 C for 1 min, followed by a nal extension at 72 C for 7 min.…”

Clinicopathological significance of fibroblast growth factor 1 in non–small cell lung cancer

“…More recently, several studies indicate that specific FGFs and FGF receptors (FGFRs), such as FGF2, FGF9, FGFR1, FGFR2, and FGFR3, are overexpressed or mutated in NSCLC cells and associated with poor prognosis of patients [6][7][8][9]. Similar with FGF2 and FGF9, FGF1 also is identified as a candidate cancer biomarker [10]. However, as far as we know, there is no study focused on the expression and function of FGF1 in NSCLC.…”

Humanization of fibroblast growth factor 1 single‐chain antibody and validation for its antitumorigenic efficacy in breast cancer and glioma cells

“…Revealing the working mechanism of hscFv1C9 is needed to promote its medical possibility. We had previously shown that the intracellular expressed mscFv1C9 arrested cell cycles via inhibiting the FGF‐1 intracrine pathway 12, 13. However, FGF‐1 works in both intracrine and paracrine pathways.…”

“…Next, we tested the effects of hscFv1C9 in cancer cells. As we had previously tested mscFv1C9 in breast cancer cells, 12,13 MDA-MB-231 was firstly used in cell proliferation and cell cycle tests.…”