Abstract
UCN-01 (7-hydroxy-staurosporine) is a potent and selective inhibitor of protein kinase C (PKC), one of several protein kinases examined. UCN-01 itself was shown to exhibit antitumor activity in vitro and in vivo in oncogene-activated human and murine tumor cell lines. Since the mechanism(s) of action of UCN-01 is thought to be different from those of alkylating agents, including mitomycin C (MMC), we tested the combined effect of UCN-01 with MMC on human epidermoid carcinoma A431 cells. UCN-01 potentiated the antiproliferative activity of MMC and yet it did not affect the growth of the cells in vitro. However, other nonselective protein kinase inhibitors, such as staurosporine, K-252a, KT6124 (a derivative of K-252a) and H7, did not enhance the activity of MMC. Isobologram analysis revealed that the interaction of UCN-01 with MMC was synergistic in its antiproliferative activity. A DNA histogram of A431 cells treated with both UCN-01 and MMC showed a block in the cell cycle at the G1/S phase. However, a histogram of cells treated with UCN-01 or MMC alone showed a G1 or a G2M block, respectively. The combined effect of UCN-01 with MMC was further examined in vivo in xenografted A431 cells in nude mice. The combination of both drugs in a single i.v. injection exhibited greater antitumor activity than MMC and UCN-01 alone (P<0.01). This synergistic antitumor effect was also confirmed in two other solid tumor cell lines, i.e. human xenografted colon carcinoma Co-3 and murine sarcoma 180. The same was observed in the i.v.-inoculated P388 leukemia model, in which we saw an increased lifespan of mice when UCN-01 was combined with MMC. These results suggests the feasibility of using UCN-01 in clinical oncology, especially in combination with alkylating agents such as MMC. In addition, this combination therapy might be a novel chemotherapeutic approach to MMC-insensitive tumors in clinical trials.
Similar content being viewed by others
References
Akinaga S, Gomi K, Morimoto M, Tamaoki T, Okabe M (1991) Antitumor activity of UCN-01, a selective inhibitor of protein kinase C, in murine and human tumor models. Cancer Res 51: 4888
Akinaga S, Ashizawa T, Gomi K, Ohno H, Morimoto M, Murakata C, Okabe M (1992) Antitumor activity of KT6124, a novel derivative of protein kinase inhibitor K-252a, and its mechanism of action. Cancer Chemother Pharmacol 29: 266
Akinaga S, Nomura K, Gomi K, Okabe M (1992) Synergistic antitumor effect of UCN-01, a protein kinase (C) inhibitor, combined with various anti-cancer agents. Proc Am Assoc Cancer Res 29: 3072
Barlogie B, Drewinko B (1980) Lethal and cytokinetic effects of mitomycin C on cultured human colon cancer cells. Cancer Res 40: 1973
Basu A, Lazo JS (1992) Sensitization of human cervical carcinoma cells to cis-diamminedichloroplatinum(II) by bryostatin 1.ibid 52: 3119
Basu A, Teicher BA, Lazo JS (1990) Involvement of protein kinase C in phorbol ester-induced sensitization of HeLa cells to cis-diamminedichloroplatinum(II). J Biol Chem 265: 8451
Basu A, Kozikowski AP, Sato K, Lazo JS (1991) Cellular sensitization of cis-diamminedichloroplatinum(II) by novel analogues of protein kinase C activators lyngbyatoxin A. Cancer Res 51: 2511
Berenbaum MC (1989) What is synergy? Pharmacol Rev 41: 93
Bush JA, Long BH, Catino JJ, Brander WT, Tomita K (1987) Production and biological activities of rebeccmycin, a novel antitumor agent. J Antibiot 40: 666
Dorr RT, Bowden GT, Alberts DS, Liddil JD (1985) Interaction of mitomycin C with mammalian DNA detected by alkaline elution. Cancer Res 45: 3510
Fine RL, Patel J, Chabner BA (1988) Phorbol esters induce multidrug resistance in human breast carcinoma cells. Proc Natl Acad Sci USA 85: 582
Geran RI, Greenberg NH, MacDonald MM, Schumacher AM, Abbot BJ (1972) Protocols for screening chemical agents and natural products against animal tumors and other biological activities. Cancer Chemother Rep 3: 1
Giard DJ, Aaronson SA, Todaro GJ, Arnstein P, Kersey JH, Dosik H, Parks WP (1973) In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst 51: 1417
Hidaka H, Inagaki M, Kawamoto S, Sasaki Y (1984) Isoquinoline-sulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinases and protein kinase C. Biochemistry 23: 5036
Hoffman J, Doppler W, Jakob A, Maly K, Posch L, Uberall F, Grunicke HH (1988) Enhancement of the antiproliferative effect of cis-diamminedichloroplatinum(II) and nitrogen mustard by inhibitors of protein kinase C. Int J Cancer 42: 382
Hoffman J, Uberall F, Posch L, Maly K, Hermann DBJ, Grunicke H (1989) Synergistic enhancement of the antiproliferative activity of cis-diamminedichloroplatinum(II) by the ether lipid analogue BM41 440, an inhibitor of protein kinase C. Lipids 24: 312
Isonishi S, Andrews PA, Howell SB (1990) Increased sensitivity to cis-diamminedichloroplatinum(II) in human ovarian carcinoma cells in response to treatment with 12-O-tetradecanoylphorbol 13-acetate. J Biol Chem 265: 3632
Kase H, Iwahashi K, Nakanishi S, Matsuda M, Yamada K, Takahashi M, Murakata C, Sato A, Kaneko M (1987) K-252 compounds, novel and potent inhibitors of protein kinase C and cyclic nucleotide-dependent protein kinases. Biochem Biophys Res Commun 142: 436
Morimoto M, Ashizawa T, Ohno H, Azuma M, Kobayashi E, Okabe M, Gomi K, Kono M, Saitoh Y, Kanda Y, Arai H, Sato A, Kasai M, Tusruo T (1991) Antitumor activity of 7-N{{2-{[2-(γ-L-glutamyl-amino)ethyl]dithio}ethyl}}-mitomycin C. Cancer Res 51: 110
Takahashi I, Saitoh Y, Yoshida M, Sano H, Nakano H, Morimoto M, Tamaoki T (1989) UCN-01 and UCN-02, a new selective inhibitors of protein kinase C: II. Purification, physico-chemical properties, structural elucidations and biological activities. J Antibiot 42: 571
Takahashi I, Kobayashi E, Nakano H, Murakata C, Saitoh H, Suzuki K, Tamaoki T (1990) Potent selective inhibition of 7-O-methyl UCN-01 against protein kinase C. J Pharmacol Exp Ther 255: 1218
Tamaoki T, Nomoto T, Takahashi I, Kato Y, Morimoto M, Tomita F (1986) Staurosporine, a potent inhibitor of phospholipid/Ca2+-dependent protein kinase. Biochem Biophys Res Commun 135: 397
Tomasz M, Lipman R, Chowdary D, Pawlak J, Verdine GL, Nakanishi K (1987) Isolation and structure of a covalent cross-link adduct between mitomycin C and DNA. Science (Washington DC) 235: 1204
Yu G, Ahmad S, Aquino A, Fairchild CR, Trepel JB, Ohno S, Suzuki K, Tsuruo T, Cowan KH, Glazer RI (1991) Transfection with protein kinase Cα confers increased multidrug resistance to MCF-7 cells expressing P-glycoprotein. Cancer Commun 3: 181
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Akinaga, S., Nomura, K., Gomi, K. et al. Enhancement of antitumor activity of mitomycin C in vitro and in vivo by UCN-01, a selective inhibitor of protein kinase C. Cancer Chemother. Pharmacol. 32, 183–189 (1993). https://doi.org/10.1007/BF00685833
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00685833