Antiproliferative effect of a lectin- and anti-Thy-1.2 antibody-targeted HPMA copolymer-bound doxorubicin on primary and metastatic human colorectal carcinoma and on human colorectal carcinoma transfected with the mouse Thy-1.2 gene.
The aim of this study was to compare the potential of two plant lectins [peanut agglutinin (PNA) and wheat germ agglutinin (WGA)], monoclonal antibody (anti-Thy-1.2), its F(ab')(2) fragments, and galactosamine as targeting moieties bound to the polymer drug carrier to deliver a xenobiotic, doxorubicin, to selected cancer cell lines. We have used primary (SW 480, HT 29) and metastatic (SW 620) human colorectal cancer cell lines and a transfectant, genetically engineered SW 620 cell line with mouse gene Thy-1.2 (SW 620/T) to test the possibility of marking human cancer with xenogeneic mouse gene and use it for effective site-specific targeting. The targeting moieties and doxorubicin were conjugated to a water-soluble copolymer based on N-(2-hydroxypropyl)methacrylamide (HPMA) acting as a carrier responsible for controlled intracellular release of the targeted drug. FACS analysis showed a strong binding of WGA-FITC to all tested cell lines. Binding of PNA-FITC was considerably weaker. The in vitro antiproliferative effect of lectin-targeted HPMA carrier-bound doxorubicin evaluated as [(3)H]TdR incorporation reflected both the intensity of the binding and the different sensitivity of the tested cancer cells lines to doxorubicin. The antiproliferative effect of conjugates targeted with WGA was comparable to that with the conjugates targeted with the anti-Thy-1.2 monoclonal antibody or their F(ab')(2) fragments. The magnitude of the cytotoxic effect of HPMA-doxorubicin targeted with PNA was lower in all tested cell lines. While the conjugates with WGA were more cytotoxic, the conjugates with PNA were more specific as their binding is limited to cancer cells and to the sites of inflammation. Noncytotoxic conjugates with a very low concentration of doxorubicin and targeted with PNA, anti-Thy-1.2, or their F(ab')(2) fragments exerted in some lines (SW 480, SW 620) low mitogenic activity. The Thy-1.2 gene-transfected SW 620 metastatic colorectal cancer cell line was sensitive to the antiproliferative effect of Thy-1.2-targeted doxorubicin as was shown for the Thy-1. 2(+) EL4 cell line and for Thy-1.2(+) concanavalin A-stimulated mouse T lymphocytes. These results represent the first indication of the suitability of transfection of human cancer cells with selected targeting genes for site-specific therapy of malignancies.
Ríhová
B
B
Institute of Microbiology, Academy of Sciences of the Czech Republic, 142 20 Prague 4-Krc, Czech Republic. Rihova@biomed.cas.cz
Jelínková
M
M
Strohalm
J
J
St'astný
M
M
Hovorka
O
O
Plocová
D
D
Kovár
M
M
Dráberová
L
L
Ulbrich
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000633:toxicity*; D002455:Cell Division / Q000187:drug effects*; D015179:Colorectal Neoplasms; D004317:Doxorubicin / Q000031:analogs & derivatives / Q000633:toxicity*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000633:toxicity; D007202:Indicators and Reagents; D008213:Lymphocyte Activation; D008689:Methacrylates; D051379:Mice; D019887:Peanut Agglutinin; D011994:Recombinant Proteins / Q000276:immunology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D018800:Thy-1 Antigens / Q000235:genetics / Q000276:immunology*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014909:Wheat Germ Agglutinins
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007202:Indicators and Reagents; D008689:Methacrylates; D019887:Peanut Agglutinin; D011994:Recombinant Proteins; D018800:Thy-1 Antigens; D014909:Wheat Germ Agglutinins; D004317:Doxorubicin; C032802:hydroxypropyl methacrylate