Abstract
CD147 is expressed at low levels in normal tissues but frequently highly expressed in a wide range of tumor types such as lung, breast, and liver and therefore it is a potentially unique therapeutic target for these diverse tumor types. We previously generated a murine antibody HAb18 which suppresses matrix met al.loproteinase-2 and matrix metalloproteinase-9 secretion, attenuates cell invasion by blocking the CD147 molecule in tumor cells. Here, we generated a chimeric antibody containing the variable heavy and variable light chains of murine HAb18 and the constant regions of human IgG1γ1 and human κ chain as a potential therapeutic agent (designated cHAb18). Quantitative measurement of cHAb18 antibody affinity for antigen CD147 with surface plasmon resonance showed the equilibrium dissociation constant KD was 2.66 × 10−10 mol/L, similar to that of KD 2.73 × 10−10 mol/L for murine HAb18. cHAb18 induced antibody-dependent cell-mediated cytotoxicity in two hepatocellular carcinoma cell lines, SMMC-7721 and Huh-7 cells. It inhibited cancer invasion and migration in hepatocellular carcinoma cells by specifically blocking CD147. Except for the depression of matrix metalloproteinase-2 and matrix metalloproteinase-9 expressions, cHAb18 antibody suppressed cell motility by rearrangement of actin cytoskeleton, which was probably induced by decreasing the phosphorylation of focal adhesion kinase, phosphatidylinositide-3 kinase (PI3K), Akt, and Girdin in the integrin signaling pathway. In an orthotopic model of hepatocellular carcinoma in BALB/c nude mice, cHAb18 treatment effectively reduced the tumor metastasis in liver and prolonged the survival. These findings reveal new therapeutic potential for cHAb18 antibody targeting CD147 on tumor therapy.
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Abbreviations
- ADCC:
-
Antibody-dependent cell-mediated cytotoxicity
- BrdU:
-
Bromodeoxyuridine
- DAPI:
-
4′,6-diamidino-2-phenylindole
- DHFR:
-
Dihydrofolate reductase
- ECM:
-
Extracellular matrix
- EGFR:
-
Epidermal growth factor receptor
- EMMPRIN:
-
Extracellular matrix metalloproteinase inducer
- FAK:
-
Focal adhesion kinase
- FBS:
-
Fetal bovine serum
- FITC:
-
Fluorescein isothiocyanate
- HCC:
-
Hepatocellular carcinoma
- H&E:
-
Hematoxylin and eosin
- HRP:
-
Horseradish peroxidase
- GHT:
-
Hypoxanthine and thymidine
- GIV/Girdin:
-
Gα-interacting vesicle-associated protein
- MMPs:
-
Matrix metalloproteinases
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- PI3K:
-
Phosphatidylinositide-3 kinase
- VH:
-
Variable heavy
- VL:
-
Variable light
- SDS:
-
Sodium dodecyl sulfate
- SPR:
-
Surface plasmon resonance
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Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (No. 81172144) and the National Science and Technology Major Project (Nos. 2012ZX10002-015 and 2012AA020806).
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Yuan Wang and Lin Yuan contributed equally to this work.
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Wang, Y., Yuan, L., Yang, XM. et al. A chimeric antibody targeting CD147 inhibits hepatocellular carcinoma cell motility via FAK-PI3K-Akt-Girdin signaling pathway. Clin Exp Metastasis 32, 39–53 (2015). https://doi.org/10.1007/s10585-014-9689-7
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DOI: https://doi.org/10.1007/s10585-014-9689-7