caa a22 4500 397561377 CHVBK 20180308164816.0 cr unu---uuuuu 161202e19960403xx s 000 0 eng 10.1093/jnci/88.7.419 doi (NATIONALLICENCE)oxford-10.1093/jnci/88.7.419 Antitumor Effect of c-myc Antisense Phosphorothioate Oligodeoxynucleotides on Human Melanoma Cells In Vitro and in Mice [Elektronische Daten] [Carlo Leonetti, Igea D' Agnano, Francesco Lozupone, Alessandra Valentini, Tim Geiser, Gerald Zon, Bruno Calabretta, Gennaro Citro, Gabriella Zupi] Background Phosphorothioate oligodeoxynucleotides ([S]ODNs) contain a modified internucleoside phosphate backbone. Antisense [S] ODNs targeted to specific oncogenes have been used with some therapeutic success in animal models of human leukemia; however, the potential for antisense [S]ODN treatment of solid tumors has only recently been explored. Purpose We evaluated the effects of antisense [S]ODNs targeted to the c-myc oncogene on the proliferation of human melanoma cells in vitro and on the growth of human melanoma xenografts in CD-1 nude (nu/nu) mice. Methods The effects of 15-mer [S]ODNs containing c-mycsense, c-myc antisense, and two different scrambled sequences on the proliferation and viability of cultures of three established human melanoma cell lines (M14, JR8, and PLF2) were determined by measuring cell numbers and use of the trypan blue exclusion test. The induction of apoptosis in these cells following treatment with [S]ODNs was evaluated by fluorescence-activated cell sorter (FACS) analysis. FACS analysis was also used to determine the effects of [S]ODN treatment on the proliferation of primary cultures of a human melanoma explant (NG cells). The expression of cMyc protein in cultured NG cells after treatment with [S]ODNs was examined by western blot analysis. The antitumor activity and the toxic effects of several [S]ODN treatment regimens were monitored by measuring differences in tumor weight (percent tumor weight inhibition), tumor growth rate (tumor growth inhibition), animal lifespan (percent increase in lifespan), the number of toxic deaths, and the median number of lung metastases in treated and control mice bearing NG xenografts. c-Myc protein expression in NG tumor cells following [S]ODN treatment was evaluated by FACS analysis, and the extent of apoptosis in these cells was determined by FACS analysis and morphologic examination. Results Treatment with antisense [S]ODNs, but not the others, inhibited the growth of all tested melanoma cultures in vitro; FACS analysis revealed that growth inhibition was associated with the induction of apoptosis. Antisense [S]ODN treatment also led to reduced cellular levels of c-Myc protein. In vivo, [S]ODN antitumor activity and toxicity were dose and schedule dependent; however, only antisense [S]ODNs exhibited antitumor activity. Mice bearing NG xenografts treated with antisense [S]ODNs showed a marked inhibition of tumor growth, a reduction in the number of lung metastases, and an increase in lifespan. Reduced levels of c-Myc protein and increased levels of apoptosis were also observed in NG tumor cells following antisense [S]ODN treatment. Conclusions Treatment of human melanoma cells and solid tumors with antisense [S]ODNs targeted to c-myc inhibits their growth and is associated with the induction of apoptosis. [J Natl Cancer Inst 1996;88:419-29] © Oxford University Press ARTICLE nationallicence Leonetti Carlo Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut D' Agnano Igea Institute of Biomedical Technology Rome aut Lozupone Francesco Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut Valentini Alessandra Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut Geiser Tim Lynx Therapeutics Inc. Hayward, CA aut Zon Gerald Lynx Therapeutics Inc. Hayward, CA aut Calabretta Bruno Thomas Jefferson University, Jefferson Cancer Institute Philadelphia, PA aut Citro Gennaro Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut Zupi Gabriella Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut JNCI: Journal of the National Cancer Institute Oxford University Press 88/7(1996-04-03), 419-429 0027-8874 88:7<419 1996 88 jnci https://doi.org/10.1093/jnci/88.7.419 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/jnci/88.7.419 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Leonetti Carlo Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut NATIONALLICENCE 700 1- D' Agnano Igea Institute of Biomedical Technology Rome aut NATIONALLICENCE 700 1- Lozupone Francesco Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut NATIONALLICENCE 700 1- Valentini Alessandra Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut NATIONALLICENCE 700 1- Geiser Tim Lynx Therapeutics Inc. Hayward, CA aut NATIONALLICENCE 700 1- Zon Gerald Lynx Therapeutics Inc. Hayward, CA aut NATIONALLICENCE 700 1- Calabretta Bruno Thomas Jefferson University, Jefferson Cancer Institute Philadelphia, PA aut NATIONALLICENCE 700 1- Citro Gennaro Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut NATIONALLICENCE 700 1- Zupi Gabriella Experimental Chemotherapy Laboratory, Regina Elena Cancer Institute Rome, Italy aut NATIONALLICENCE 773 0- JNCI: Journal of the National Cancer Institute Oxford University Press 88/7(1996-04-03), 419-429 0027-8874 88:7<419 1996 88 jnci Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford