Inhibition of 12-O-Tetradecanoylphorbol~13

[CANCER RESEARCH 42, 2841-2845, July 1982] 0008-5472/82/0042-0000$02.00 Inhibition of 12-O-Tetradecanoylphorbol~13-acetate-induced Epidermal Ornithine Decarboxylase Activity by Phospholipase A2 Inhibitors and Lipoxygenase Inhibitor Teruo Nakadate, ' Satoshi Yamamoto, Miki Ishii, and Ryuichi Kato

Department of Pharmacology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160, Japan

ABSTRACT the TPA-induced elevation of mouse epidermal ODC. The bio chemical mechanism of induction of ODC by TPA, however, Application of 12-O-tetradecanoylphorbol-IS-acetate (TPA; has not yet been fully understood. 20 nmol/mouse), a tumor-promoting agent, to mouse skin Recently, TPA has been shown to enhance the release of results in an induction of epidermal ornithine decarboxylase prostaglandins and/or arachidonic acid in dog kidney (MDCK) (ODC; EC 4.1.1.17). Induction of ODC by TPA was inhibited by cells (9, 10, 24), C3H/10T'/2 mouse and chick embryo fibro- treatment of skin with indomethacin (1.12 /xmol/mouse), a blasts (19, 20), rabbit leukocytes (8), Friend erythroleukemia cyclooxygenase inhibitor, and the ODC activity suppressed by cells (39), epidermal cells (4), and epidermis (1, 3, 30). Verma indomethacin was completely restored by concurrent applica ef al. (30, 33) reported that pretreatment of prostaglandin tion of prostaglandin E? (PGE2) (140 nmol/mouse) as de synthesis inhibitors such as indomethacin, flufenamic acid, or scribed first by Vermaef a/. (Cancer Res., 40. 308-315,1980). acetylsalicylic acid markedly inhibits the induction of epidermal Treatment of mice with tetracaine (20 and 100 jiimol/mouse), ODC by TPA. The inhibition of TPA-induced ODC by indometh a nonspecific phospholipase A2 inhibitor, inhibited the induction acin was completely overcome by the treatment with PGE2. of ODC by TPA. More specific phospholipase A2 inhibitors, They suggested that PGE2 may play a crucial role in TPA- mepacrine (20 Â¡nmol/mouse) and p-bromophenacyl bromide induced elevation of ODC activity. (10 /imol/mouse), also inhibited the ODC induction. The TPA- However, our preliminary data demonstrated that TPA-in induced ODC inhibited by mepacrine was not restored by the duced ODC activity of mouse epidermis was not restored by treatment of mice with PGE2. TPA-induced ODC inhibited by PGE2 when the ODC induction was suppressed by mepacrine, either mepacrine or p-bromophenacyl bromide was partially a phospholipase A2 inhibitor. The above finding prompted us but significantly restored by treatment with arachidonic acid (1 to investigate the possible involvement of phospholipase A2 to 40 ftmol/mouse). Neither PGE2 nor arachidonic acid alone activation and arachidonate cascade metabolites in the mech could induce the epidermal ODC. Treatment of mice with anism of ODC induction by TPA in mouse epidermis. nordihydroguaiaretic acid (10 to 90 /umol/mouse), a lipoxygen- The results presented here suggest that phospholipase A2 ase inhibitor, also inhibited the induction of ODC by TPA. These stimulation and resultant arachidonate metabolites, i.e., not results strongly indicate that the stimulation of phospholipase only cyclooxygenase product(s) but also lipoxygenase prod- A2 activity is a crucial process in inducing mouse epidermal uct(s), are involved in the mechanism of ODC induction by TPA. ODC by TPA and not only cyclooxygenase product (i.e., PGE2) but also lipoxygenase product(s) are involved in the mechanism MATERIALS AND METHODS of ODC induction. Our present data also suggest that the above arachidonate metabolites are essential but not sufficient factors Female CD-1 mice (Charles River, Kanagawa-ken, Japan), 7 to 8 for the TPA-stimulated induction of ODC. weeks old, were used. The dorsal hair of each mouse was shaved with clippers at least 2 days before use, and only those mice showing no INTRODUCTION hair regrowth were used. All animals received water ad libitum through out the experimental period. Mice were routinely sacrificed between 4 ODC2 (EC 4.1.1.17; L-ornithine carboxy-lyase), the rate-lim and 6 p.m. All chemicals applied to mouse skin were dissolved in iting enzyme in the polyamine-biosynthetic pathway, is elevated reagent grade acetone except for lysophosphatidylcholine which was dissolved in ethanol:water (5.1:1). The chemicals were applied to the in various proliferative cell systems and implicated in neoplastic shaved area of individual mice in a total volume of 0.2 ml using a growth (12, 27, 31 ). In adult mouse epidermis, a rapid, transient micropipet for each occasion. Five hr after the TPA administration, the induction of ODC is observed by application of TPA, a potent mice were sacrificed by cervical dislocation and the skins were excised. tumor-promoting agent (21, 22). It has been suggested that an The skin was placed in 55Â°water for 30 sec, submerged in ice water induction of ODC is necessary for tumor promotion. However, for at least 30 sec (22), and then placed on an ice-cold plate. Epidermis recent findings indicate that ODC induction is essential but not was removed free from dermis and fat by scraping with a razor blade. a sufficient factor for the mechanism of skin tumor promotion The epidermis was immediately refrigerated by liquid nitrogen and stored at - 70Â°until just prior to homogenization. The epidermal sheets (14, 15). It was reported that pretreatment with colchicine or other microtubule-disrupting agents (23), local anesthetics were homogenized in 20 mw Tris-HCI buffer, pH 7.2, in a Polytron PT- 10 homogenizer for 10 to 15 sec at 4Â°. The supernatant fraction (41 ), retinoids (11, 31, 34, 35), and putrescine (38) suppressed obtained after centrifugation at 30,000 x g for 30 min at 0Â°was used 1 To whom requests for reprints should be addressed. for estimation of enzyme activities. In most cases, a soluble epidermal 2 The abbreviations used are: ODC, ornithine decarboxylase; TPA, 12-O- extract was prepared from one mouse. ODC activity was determined tetradecanoylphorbol-13-acetate; PGE2. prostaglandin E2; NDGA, nordihydo- by measuring the release of MCO2 from DL-fJ-""C]ornithine as de guaiaretic acid; BPB, p-bromophenacyl bromide. Received December 10, 1981; accepted April 1, 1982. scribed by Russell and Snyder (25). In brief, incubation was carried

JULY 1 982 2841

out for 60 min at 37Â° in a medium containing 0.4 /Â¿molpyridoxal shown to be a phospholipase A2 inhibitor (2, 16, 17, 29, 40). phosphate, 1.0 /imol dithiothreitol, 0.2 fimol L-ornithine, 30 fimo\ Tris BPB, another phospholipase A2 inhibitor with an inhibitory base (pH 7.2), 0.5 ml epidermal extract, and approximately 0.5 fiC\ DL-[1-MC]ornithine (51.3 mCi/mmol) in a final volume of 2.0 ml. In mechanism that is different from that of mepacrine (17, 28, 36), also inhibited the TPA-induced increase in epidermal ODC order to absorb the generated CO2 and '"CO2, Hyamine 10X was (Chart 1). The inhibition of TPA-induced ODC by mepacrine placed in a central well of the incubation flask. The reaction was stopped by the addition of 1.0 ml 2 M citric acid, and incubation was could not be reversed by treatment of mice with 140 nmol of continued for an additional 30 min to ensure complete absorption of PGE2 (Chart 2). The above dose of PGE2 completely restored MCO2. Hyamine solution was transferred into a scintillation vial con the TPA-induced ODC suppressed by indomethacin (see Table taining 10 ml of a toluene-based scintillation cocktail. Radioactivity 1). Further increase in doses of PGE2 also failed to counteract was measured in a liquid scintillation counter (Aloka, Tokyo, Japan). the inhibitory effect of mepacrine (Chart 2). Counting efficiency was approximately 87%. All enzyme activities were Topical application of arachidonic acid significantly restored corrected against the value which was measured in the presence of an the TPA-induced ODC which was suppressed by mepacrine excess amount of a-methylornithine (final 5 mM), a reversible inhibitor of ODC. Activity was expressed as nmol CO2 in 60 min per mg protein. Enzyme activity was proportional to the amount of extract and to the tetracaineD time of incubation within our experimental conditions. None of the mepacrine BBPBLiLU drugs which we used (indomethacin, tetracaine, mepacrine, BPB, arachidonic acid, or NDGA) directly interfered with the ODC assay 100 1IiÃjm system. The protein concentration of the epidermal extract was measured by the method of Lowry et al. (13), with bovine serum albumin as standard. TPA, indomethacin, mepacrine, arachidonic acid NDGA, and 50 L-a-lysophosphatidylcholine were purchased from Sigma Chemical Co., St. Louis, Mo.; a-methylornithine was from Calbiochem-Behring Corp., La Jolla, Calif., BPB was from Wako Pure Chemical Industries Ltd., Osaka, Japan. PGE2 was kindly donated by the Research Institute inhibitor 0 5 10 20 100 of Toray Co., Kamakura, Japan. DL-[1-"'C]Ornithine (51.3 mCi/mmol) (jjmol) was obtained from New England Nuclear, Boston, Mass. Chart 1. Effect of tetracaine, mepacrine, and BPB on the TPA-induced epi dermal ODC activity. Results of 3 separate experiments are shown. In Experiment 1, mice were treated with 20 nmol of TPA or TPA plus tetracaine (20 or 100 RESULTS /irnol). Two hr after TPA treatment, the same amount of tetracaine was applied to the mouse skin. In Experiment 2 or 3, mice were treated with mepacrine (5, 10, A single topical application of TPA (20 nmol/mouse) resulted or 20 /imol) or BPB (5, 10, or 20 /imol), respectively. 30 min prior to the application of TPA (20 nmol). The same amounts of mepacrine or BPB were in a substantial and transient increase in mouse epidermal ODC as reported by O'Brien ef al. (21, 22). As shown in Table 1, applied again to mouse skin concurrently with TPA. In all experiments, mice were sacrificed for determination of ODC activity 5 hr after TPA treatment. Results are treatment of mouse skin with indomethacin (1.12 jumol/mouse expressed as the percentage of ODC activity induced by TPA in the presence of inhibitor relative to the ODC activity induced by TPA alone. In fact, we obtained for 2 treatments) resulted in a suppression of ODC induction original data as mean Â±S.E. (A Â±a) of individual determinations from 5 mice. by TPA. Inhibition of TPA-induced ODC by indomethacin was Each result (mean Â±S.E. = C Â±cS.E.) was calculated by the equation:

counteracted by PGE? (140 nmol/mouse) which was applied C-A/B x 100 concurrently with indomethacin treatments. These results are consistent with those of Verma ef at. (30, 33). C - A/B [(a/A? + (b/B)2] x 100 When tetracaine (20 and 100 /Â¿mol/mouse), a local anes in which B Â±b is control value (TPA alone). â€¢,p < 0.05 versus TPA alone; â€¢â€¢.p thetic, was applied to the mouse skin, TPA-induced ODC was < 0.01 versus TPA alone. suppressed dose responsively (Chart 1). As shown in Chart 1, ODC induction by TPA was also inhibited dose responsively by the treatment of mouse skin with mepacrine which has been I j il O Table 1 Inhibitory effect of indomethacin on TPA-induced epidermal ODC activity and the restorative effect of PGÂ£2on the indomethacin-caused inhibition Acetone (vehicle), 1.12 /imol of indomethacin, or indomethacin plus 140 nmol of PGE2 was applied to mice skin 2 hr prior to the topical application of 20 nmol of TPA. The same amounts of indomethacin (1.12 /imol) and/or PGE2 (140 nmol) were applied again to the mice concurrently with TPA. Mice were sacrificed 5 hr after TPA treatment, and ODC activity was determined by using soluble epidermal Ã± Ã±Ã±Ã± extracts. activity (rifa)2 - O KO 280 560 (nmol COj/60 PretreatmentAcetone min/mgprotein)0.13 Chart 2. Effect of PGE2 on inhibition of the induction of ODC activity by Â±0.08a mepacrine. Mice were treated with acetone (vehicle), 20 /imol of mepacrine, or mepacrine plus 140, 280, and 560 nmol of PGE2 30 min prior to the topical Acetone TPA 1.07 Â±0.08 0.41 Â±0.03b application of 20 nmol of TPA. The same amounts of mepacrine (20 /imol) and/ Indomethacin TPA + indomethacin 1.05 Â±0.21Â° or PGE2 (140 to 560 nmol) were applied again to mouse skin concurrently with Indomethacin + PGE2TreatmentAcetoneTPA + indomethacin + PGE2ODC TPA. Mice were sacrificed for determination of ODC activity 5 hr after TPA * Mean Â±S.E. of individual determinations from 7 or 8 mice. treatment. Each value is the mean of individual determinations from 5 mice; bars, 6 p < 0.01 versus TPA. S.E. â€¢,p< 0.01 versus TPA; +, p < 0.01 versus TPA and p is not significant c p < 0.01 versus TPA + indomethacin; p is not significant versus TPA. versus TPA plus mepacrine.

2842 CANCER RESEARCH VOL. 42

(20 jumol/mouse), to the extent of 28% with 10 jumol and 34% with 40 /Â¿molofarachidonlc acid (Table 2). Further increase in Â§â€¢8ARBOXYLASE doses of arachidonic acid failed to complete the restoration. OuÂ»gUJQj|051"NDGA Arachidonic acid(1 and 10/Â¿mol/mouse) also partially restored the TPA-induced ODC which was inhibited by BPB (10 /Â¿mol/ mouse) (Table 2). In addition, topical application of lysophosphatidylcholine (40 fimol/mouse) significantly enhanced the ft30 restoring effect of arachidonic acid (10 /Â¿mol/mouse) when the TPA-induced ODC was suppressed by BPB and also tended to enhance the restoring effect of arachidonic acid (40 /Â¿mol/ â€”i-I0i1I3i10ft mouse) when TPA-induced ODC was suppressed by mepacrine 90 (Table 2). Uimol)i Either PGE2 (140 nmol/mouse) or arachidonic acid (40 Chart 3. Effect of NDGA on TPA-induced epidermal ODC activity. Mice were /Â¿mol/mouse) alone could not induce the epidermal ODC (Table treated with acetone (vehicle) or 1 to 90 /Â¿molof NDGA 40 min prior to the application of 20 nmol of TPA. The same amounts of NDGA (1 to 90 fimol) were 3). applied again to mouse skin concurrently with TPA. Mice were sacrificed for the NDGA, an inhibitor of lipoxygenase (7, 18, 37), inhibited the determination of ODC activity 5 hr after TPA treatment. Each value is the mean ODC induction by TPA in a dose-dependent manner (Chart 3). of individual determinations from 5 or 10 mice; bars, S.E. â€¢,p< 0.01 versus TPA. Table 2 Effects of arachidonic acid and lysophosphatidylcholine on inhibition of TPA- DISCUSSION induced epidermal ODC activity by mepacrine and BPB Mice were treated with acetone (vehicle), mepacrine (20 /Â¿mol),or BPB (10 Our present data clearly show that the TPA-induced ODC /imol) 30 min prior to the topical application of TPA (20 nmol) or TPA plus arachidonic acid (1 to 40 /imol). The same amounts of mepacrine (20 /imol) or suppressed by indomethacin, a cyclooxygenase inhibitor, was BPB (10 (Â¿mol)were applied again to the mice skin concurrently with TPA. When completely restored by the concurrent application of PGE2 with mice were treated with lysophosphatidylcholine, either lysophosphatidylcholine indomethacin. The results are consistent with those of Verma or vehicle (ethanohwater, 5.1:1) was applied on mouse skin in a volume of 100 fil 15 to 30 min after TPA. Mice were sacrificed for the determination of ODC ef al. (30, 33). TPA has been shown to enhance the release of activity 5 hr after TPA treatment. prostaglandins, such as PGE2, in various cell types including activity epidermal cells (4) and epidermis (1, 3, 30). Thus, we con (nmol CO2/60 firmed the previous findings (30) that PGE2 plays an essential PretreatmentExperiment min/mgprotein)0.06 role in the mechanism of ODC induction by TPA in mouse 1AcetoneAcetoneMepacrineMepacrineMepacrineMepacrineTreatmentAcetoneTPATPA Â±0.05a1.19 epidermal cells. Â±0.200.09 Recently, FÃ¼rstenberger ef al. (4) reported that TPA stimu Â±O.OI60.40 +mepacrineTPA Â±0.05e0.46 lates the release of arachidonic acid as well as prostaglandins + mepacrine + arachi from epidermal cells prelabeled with ['"C]arachidonic acid. The (10/imol)TPAdonic acid + mepacrine + arachi Â±0.06C0.65 above findings indicate that as with other types of cells, also in il)TPAdonic acid (40 ni'Â« epidermal cells, TPA releases arachidonic acid by stimulating + mepacrine + arachi Â±0.17 donic acid (40 Â¡Â¡mon+lyso phospholipase A2. If that is the case, phospholipase A2 inhibi phosphatidylcholine (20 fimol)n255554ODC tors should block the ODC induction by TPA. Our present data clearly demonstrate the inhibition of TPA-caused ODC induc Experiment 2 Acetone Acetone 2 0.07 Â± 0.02 tion by relatively specific phospholipase A2 inhibitors, i.e., Acetone TPA 4 1.36 Â± 0.06 0.16 Â± 0.03b mepacrine (2, 16, 17, 29, 40) and BPB (17, 28, 36). Tetra- BPB TPA + BPB 4 BPB TPA + BPB + arachidonic acid 4 caine, one of the local anesthetics which has been shown to (1 nmol) inhibit phospholipase A2 (26), also inhibits the ODC induction BPB TPA + BPB + arachidonic acid 4 by TPA. These results strongly suggest that phospholipase A2 (lOfimol) TPA + BPB + arachidonic acid 5 0.52 Â±0.06e BPB stimulation is involved in the mechanism of ODC induction by (10 fimol) + lysophosphati TPA. dylcholine (20 fimol) When ODC induction by TPA was inhibited by mepacrine, " Mean Â±S.E. of individual determinations from 4 to 5 mice. b p < 0.01 versus TPA. PGE2 failed to overcome the inhibitory effect of mepacrine c p < 0.01 versus TPA plus mepacrine. (Chart 2). This finding confirms the idea that PGE2 is an a p < 0.01 versus TPA plus BPB. a p< 0.05 versus TPA plus BPB plus arachidonic acid (10 fimol). essential factor but not one sufficient to induce ODC activity by TPA (30). The fact that PGE2 alone failed to stimulate ODC Table 3 induction (Table 3; Ref. 30) supports the above contention. Effect of PGEi and arachidonic acid on epidermal basal ODC activity When TPA enhances the release of arachidonic acid by stim Mice were treated with acetone (vehicle). TPA (20 nmol), PGE2 (140 nmol), or ulating phospholipase A2, part of the released arachidonic acid arachidonic acid (40 /imol) 5 hr before sacrifice. would be oxidized through the cyclooxygenase pathway to ODC activity prostaglandin derivatives or through the lipoxygenase pathway Treatment (nmol CO2/60 min/mg protein) to hydroperoxy or hydroxy derivatives of fatty acids. Partial Acetone 0.07 Â±0.12 restoration of TPA-stimulated ODC induction which was in TPA 1.15 Â±0.13 PGE2 0.08 Â±0.04 hibited by phospholipase A2 inhibitors, by exogenous arachi Arachidonic acid 1.12 Â±0.03 donic acid indicates the possible involvement of an unknown Mean Â±S.E. of individual determinations from 6 mice. arachidonate metabolite(s) other than PGE2 in the mechanism

JULY 1982 2843

of ODC induction by TPA. PGE2 fully restored the TPA-caused 9. Levine, L..andHassid, A. Effects of phorbol-12,13-diesters on prostaglandin ODC induction inhibited by indomethacin but failed to restore production and phospholipase activity in canine kidney (MDCK) cells. Bio chem. Biophys. Res. Commun., 79. 477-484, 1977. the ODC induction inhibited by mepacrine. Thus, it is likely that 10. Levine, L., and Ohuchi. K. Stimulation by carcinogens and promoters of the unknown arachidonate metabolite(s) (other than PGE2) is prostaglandin production by dog kidney (MDCK) cells in culture. Cancer a product(s) of the lipoxygenase pathway rather than of the Res., 38. 4142-4146. 1978. 11. Lichti, U.. Patterson, E., Hennings, H., and Yuspa. S. H. Differential retinoic cyclooxygenase pathway. Moreover, the present studies dem acid inhibition of ornithine decarboxylase induction by 12-O-tetradecanoyl- onstrated the partial but significant inhibition of TPA-caused phorbol-13-acetate and by germicidal ultraviolet light. Cancer Res., 41: 49- ODC induction by NDGA, which inhibits lipoxygenase (7, 18, 54, 1981. 12. Lowe, N. J. Epidermal ornithine decarboxylase, polyamines. cell prolifera 37) without inhibiting cyclooxygenase (18). These results tion, and tumor promotion. Arch. Dermatol., ) (6. 822-825, 1980. strongly indicate the involvement of a lipoxygenase product(s) 13. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem., ÃŽ93:265-275, in the mechanism of ODC induction by TPA. 1951. Arachidonic acid could partially but not completely overcome 14. Marks, F., Bertsch. S., and FÃ¼rstenberger. G. Ornithine decarboxylase the inhibitory effect of phospholipase A2 inhibitors, i.e., mepa activity, cell proliferation, and tumor promotion in mouse epidermis in vivo. Cancer Res., 39. 4183-4188. 1979. crine or BPB. When TPA stimulates phospholipase A2, lyso- 15. Marks. F., FÃ¼rstenberger. G., and Kownatzki, E. Prostaglandin E-mediated phospholipids would be accumulated in the tissue with a con mitogenic stimulation of mouse epidermis in vivo by divalent cation ionophore A 23187 and by tumor promoter 12-O-tetradecanoylphorbol-13-acetate. current release of arachidonic acid. Therefore, it is possible Cancer Res., 41. 696-702, 1981. that a certain type of lysophospholipid plays a significant role 16. Markus. H. B., and Ball, E. G. Inhibition of lipolytic processes in rat adipose in the mechanism of ODC induction by TPA. Recently, Verma tissue by antimalarial drugs. Biochim. Biophys. Acta, 787: 486-491, 1969. 17. Mitchell, S.. Poyser, N. L., and Wilson, N. H. Effect of p-bromophenacyl and Boutwell (32) reported that extracellular calcium is re bromide, an inhibitor of phospholipase A2, on arachidonic acid release and quired for the ODC induction by TPA. A certain type of lyso prostaglandin synthesis by the guinea pig uterus in vitro. Br. J. Pharmacol., phospholipid plays a significant role in calcium movement 59: 107-113, 1977. 18. Morris, H. R., Piper, P. J.. Taylor, G. W., and Tippins, J. R. The effect of through the biological membrane (5). Our preliminary data arachidonate lipoxygenase substrates and inhibitors on SRS-A release in showed that lysophosphatidylcholine could only slightly en the guinea pig lung. Br. J. Pharmacol., 66: 452P, 1979. hance the restorative effect of arachidonic acid. However, at 19. Mufson, R. A., DeFeo. D., and Weinstein, l. B. Effects of phorbol ester tumor promoters on arachidonic acid metabolism in chick embryo fibroblasts. Mol. this moment, it is too early to draw a conclusion concerning Pharmacol., 76: 569-578, 1979. the role of lysophospholipids in the mechanism of ODC induc 20. Mufson, R. A., Laskin, J. D.. Fisher, P. B., and Weinstein, I. B. Melittin shares certain cellular effects with phorbol ester tumor promoters. Nature tion by TPA. It cannot be denied that some nonspecific toxic (Lond.). 280: 72-74. 1979. effects of mepacrine and BPB prevented the full restoration of 21. O'Brien, T. G. The induction of ornithine decarboxylase as an early, possibly ODC activity by arachidonic acid. obligatory, event in mouse skin carcinogenesis. Cancer Res., 36: 2644- 2653, 1976. The fact that arachidonic acid alone failed to induce ODC 22. O Brien. T. G.. Simsiman, R. C., and Boutwell. R. K. Induction of the activity indicates that arachidonate metabolites are necessary polyamine-biosynthetic enzymes in mouse epidermis by tumor-promoting agents. Cancer Res.. 35: 1662-1670, 1975. but not sufficient for the induction of ODC. Recently, it was 23. O'Brien, T. G.. Simsiman. R. C.. and Boutwell, R. K. The effect of colchicine reported that 1,2-diacylglycerol is generated in plasma mem on the induction of ornithine decarboxylases by 12-O-tetradecanoyl-phor- branes of chick embryo myoblasts by TPA (6). Therefore, in bol-13-acetate. Cancer Res., 36: 3766-3770, 1976. 24. Ohuchi, K., and Levine, L. Stimulation of prostaglandin synthesis by tumor- addition to the stimulation of phospholipase A2, TPA may cause promoting phorbol-12,13-diesters in canine kidney (MDCK) cells. J. Biol. a variety of alternations in lipid metabolism. Thus, some other Chem., 253: 4783-4790, 1978. factor(s) such as 1,2-diacylglycerol, calcium ion, or other un 25. Russell. D. H.. and Snyder, S. H. Amine synthesis in rapidly growing tissues: ornithine decarboxylase activity in regenerating rat liver, chick embryo, and known intracellular second messenger(s), in addition to phos various tumors. Proc. Nati. Acad. Sei. U. S. A., 60: 1420-1427, 1968. pholipase A2 products, may play an important role in the 26. Scherphof, G. L.. Scarpa, A., and van Toorenenbergen. A. The effect of mechanism of ODC induction by TPA. local anesthetics on the hydrolysis of free and membrane-bound phospholipids catalyzed by various phospholipases. Biochim. Biophys. Acta, 270: 226-240, 1972. 27. Tabor, C. W., and Tabor. H. 1,4-Diaminobutane (putrescine), spermidine, REFERENCES and spermine. Annu. Rev. Biochem.. 45: 285-306, 1976. 28. Vallee, E.. Gougat, J.. Navarro, J., and Delahayes. J. F. Anti-inflammatory 1. Bresnick. E., Meunier. P., and Lamden, M. Epidermal prostaglandins after and platelet anti-aggregant activity of phospholipase A. inhibitors. J. Pharm. topical application of a tumor promotor. 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Downloaded from cancerres.aacrjournals.org on September 30, 2021. © 1982 American Association for Cancer Research. Inhibition of 12-O-Tetradecanoylphorbol-13-acetate-induced Epidermal Ornithine Decarboxylase Activity by Phospholipase A 2 Inhibitors and Lipoxygenase Inhibitor

Teruo Nakadate, Satoshi Yamamoto, Miki Ishii, et al.

Cancer Res 1982;42:2841-2845.

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