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Perillyl Alcohol as a Chemopreventive Agent in N-Nitrosomethylbenzylamine-induced Rat Esophageal Tumorigenesis¹

Department of Pathology [B. W. L.], and Division of Environmental Health Sciences, School of Public Health and Comprehensive Cancer Center [R. N., P. S. C., A. G., R. A., W. F., G. D. S.], The Ohio State University, Columbus, Ohio 43210

	ABSTRACT

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Perillyl alcohol (POH) is a monoterpene found in lavender, spearmint, and cherries. Phase I clinical trials with this agent have shown a favorable toxicity profile and preliminary data indicate some chemotherapeutic efficacy in advanced cancers. Animal studies have demonstrated the ability of POH to inhibit tumorigenesis in the mammary gland, liver, and pancreas. Although the precise mechanism of action is unclear, POH has been shown to inhibit the farnesylation of small G-proteins, including Ras, up-regulate the mannose-6-phosphate receptor, and induce apoptosis. Previous studies in our laboratory using the rat model of squamous cell carcinoma of the esophagus have shown that a specific Ha-ras codon 12 mutation is important for tumor promotion and progression. Given the limited toxicity of POH in humans, its proven efficacy in several animal models and its potential to inhibit Ha-ras farnesylation, we conducted an animal study to evaluate the efficacy of POH as a chemopreventive agent for squamous cell carcinoma of the esophagus. Male Fischer-344 rats were treated s.c. with 0.25 mg/kg b.w. of N-nitrosomethylbenzylamine three times a week for 5 weeks. Three days after the final carcinogen dose, they were started either on control diet or diets containing 0.5 or 1.0% POH. At 25 weeks, the animals were sacrificed, and esophageal tumors were counted. Animals fed either dose of POH showed a significant increase in dysplasia when compared with controls (P < 0.05) and a nonsignificant trend toward increased tumor multiplicity. Additionally, 1.0% POH did not affect Ras membrane localization. These data indicate that POH has a weakly promoting effect early in nitrosamine-induced esophageal tumorigenesis and suggest that POH may not be an effective chemopreventive agent for esophageal cancer in humans.

	INTRODUCTION

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POH³ is a monocyclic monoterpene found in essential oils of plants such as cherries, lavender, and spearmint. Animal studies with this compound have found POH to be effective in cancer prevention. Multiple studies have shown this compound to be an effective inhibitor of tumor initiation. i.p. treatment with POH 1 week before the administration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone resulted in a decrease in tumor incidence and multiplicity in the mouse lung (1) . Topical treatment with 10 mM POH significantly inhibits tumor incidence and multiplicity, average tumor size, and average tumor burden/mouse in the UVB-induced murine skin cancer model (2) . In the azoxymethane-induced rat colon cancer model, treatment with POH before, during, and after carcinogen administration resulted in a 30% decrease in tumor incidence (3) . POH has also been effective in the postinitiation stages of cancer development. In the diethylnitrosamine induced rat model of hepatocarcinogenesis, POH treatment 2 weeks after carcinogen administration resulted in a 10-fold reduction in liver tumor mass as compared with control (4) .

Chemotherapeutic efficacy has been seen in 7,12-dimethylbenz(a)anthracene and N-nitroso-N-methylurea models of rat mammary carcinogenesis; monoterpenes have caused complete regression of >80% of established mammary carcinomas (5, 6, 7, 8) . Additionally, in the transplantable pancreatic tumor model in the hamster, POH treatment caused regression in 25% of pancreatic tumors (9) . In Phase I trials, POH has a favorable toxicity profile and has shown evidence of efficacy in several patients with refractory malignancies (10 , 11) .

The mechanisms of action of POH are not fully known. Early investigations with D-limonene and POH demonstrated their ability to inhibit the isoprenylation of small G-proteins (12, 13, 14, 15) . Crowell et al. (13) demonstrated POH’s ability to selectively inhibit isoprenylation of 21–26-kDa proteins implicating the disruption of the intracellular localization of small GTP-binding proteins such as Ras as a mechanism of action. Isoprenylation of Ras is critical to its membrane association and transforming ability (16) . Inhibition of isoprenylation enzymes, farnesyl and geranylgeranyl transferases, has been demonstrated with both POH and its metabolites (14 , 17) . The doses required for this inhibition are readily attainable in vivo (5 , 18 , 19) . Studies by Ren and Gould (20) demonstrated a 17% reduction in Ras prenylation by farnesyl transferase in vivo with 2% dietary POH. However, this inhibition may not effect Ras plasma membrane association (21) and, therefore, may not be the primary mechanism of action. Other mechanisms believed to be involved include modulation of cell cycle regulator molecules such as cyclin D1 (22 , 23) , the induction of the mannose 6-phosphate receptor, and TGF-ß1 (4 , 8 , 24) , induction of apoptosis (4 , 24, 25, 26) , and modulation of AP-1 activity (2 , 27) .

Esophageal cancer ranks eighth in incidence and is the fifth most common cause of cancer deaths throughout the world (28) . Extensive epidemiological research in high incidence areas such as China and South Africa has suggested that exposure to N-nitrosamine compounds is a probable etiological factor in the development of esophageal cancer (29 , 30) . Among these, NMBA is the most potent esophageal carcinogen in rodents (31, 32, 33) . Because of its potency, organ specificity and possible role as a human carcinogen, NMBA has been used extensively to study the development, progression, and chemoprevention of esophageal tumors in rodents.

In the NMBA-induced rat model of squamous cell carcinoma of the esophagus, Ha-ras GA transition mutations have been observed in the majority of papillomas (34, 35, 36, 37) , and this mutation is believed to contribute to postinitiation events (36 , 37) . The role of ras mutation in the development of esophageal cancer provides an ideal target for the evaluation of chemopreventive agents in this model. Given POH’s ability to inhibit Ras farnesylation, its demonstrated efficacy in preventing some forms of cancer and its favorable toxicity profile, we conducted an animal study to evaluate its ability to inhibit postinitiation events in NMBA-induced squamous cell carcinoma in the rat esophagus. In this study, we found that POH treatment had a weakly promoting effect early in esophageal tumorigenesis. Additionally, doses causing toxicity to rats were not sufficient to inhibit Ras membrane association. These data suggest that POH may not be an effective chemopreventive agent for esophageal cancer.

	MATERIALS AND METHODS

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Animals.
Four-to-5-week-old male Fischer 344 rats were purchased from Harlan Sprague Dawley (Indianapolis, IN). The animals were housed under standard conditions (20 ± 2°C; 50 ± 10% relative humidity; 12 h light/dark cycles). Rats were fed a modified AIN-76A-purified diet containing 20% casein, 0.3% D,L-methionine, 52% cornstarch, 13% dextrose, 5% cellulose, 5% corn oil, 3.5% AIN salt mixture, 1.0% AIN vitamin mixture, and 0.2% choline bitartrate (Dyets, Bethlehem, PA). Food and water were provided ad libitum. Hygienic conditions were maintained by twice weekly cage changes.

Chemicals.
NMBA was purchased from Ash Stevens (Detroit, MI) and determined to have a purity >98% by high-performance liquid chromatography analysis. S(-) POH (96% pure) was purchased from Sigma-Aldrich (Milwaukee, WI).

Chemoprevention Study.
Following a two-week acclimation period, rats were randomized into experimental groups (Table 1) . Animals treated with NMBA were dosed with 0.25 mg/kg body weight of NMBA by s.c. injection three times/week for 5 weeks. Group 1 was dosed with 20% DMSO in water as a vehicle control. Three days after the final NMBA dosing, POH was added to the diets of groups 3–5. POH was mixed with AIN-76A in concentrations of 0.5 and 1.0% weekly. Previous bioassays with POH in Fisher 344 rats have yielded conflicting toxicity results. Reddy et al. (3) observed a significant decrease in body weight to occur in rats fed 0.25% POH, whereas Mills et al. (4) did not observe toxicity in rats fed 1.0–2.0% POH. Therefore, we chose to use intermediate dose levels to maximize efficacy and minimize toxicity. Food jars were changed daily, 6 days/week, to minimize the loss of POH because of volatilization. Daily food consumption and weekly weight changes were recorded. Rats were housed 3/cage, except for 6 animals in groups 2 and 5. These 12 animals were housed individually to more closely monitor individual food consumption. At 25 weeks, rats were euthanized by CO₂ asphyxiation and subjected to gross necropsy. Blood samples, liver, lung, and stomach were collected from 5 animals in groups 2, 4, and 5 to evaluate for toxicity. The esophagus of each rat was removed and opened longitudinally. Papillomas were counted and measured. Tumor size was calculated using the formula for a prolate spheroid [(length x width x height x )/6]. The esophagus was then divided into two parts. Esophageal epithelium from one part was stripped of the submucosal and muscularis layers and frozen in liquid nitrogen. Papillomas were frozen separately. The other portion was fixed in 10% neutral-buffered formalin for 4 h and transferred to PBS.

Western Blotting.
The esophageal epithelium from 6 rats in groups 2 and 5 was placed in PBS and mechanically disrupted with a tissue homogenizer. The homogenate was ultracentrifuged at 100,000 x g for 30 min. The supernatant containing cytoplasmic proteins was removed, and the pellet containing membrane bound proteins was dissolved in an equal volume of PBS. Positive control SW480 cells were prepared in the same manner. Protein concentrations were determined using the Bio-Rad protein quantification assay. Aliquots of protein (10 µg) from both the cytoplasmic and membrane bound samples from each rat, as well as from the membrane positive control, were mixed with an appropriate volume of 4x NuPage LDS sample buffer (Invitrogen, Carlsbad, CA) and heated to 70°C for 10 min. Samples were loaded on precast 1.0% Bis-Tris gel (Invitrogen) and electrophoresed with NuPage 4-morpholinepropanesulfonic acid running buffer (Invitrogen) at 200 V for 1 h. Proteins were transferred onto a polyvinylidene difluorite membrane by electroblotting for 1.5 h at 30 V using NuPage transfer buffer (Invitrogen). Membranes were evaluated for Ras protein using the WesternBreeze Novex chromogenic Western blot immunodetection kit (Invitrogen) according to the manufacturers protocol. Briefly, membranes were washed, incubated with 1 µg/ml pan-ras (Ab-3) antibody (Oncogene, Boston, MA) for 1 h, and then with antimouse secondary antibody solution for 30 min. Membranes were developed with the provided chromogenic substrate for 30 min and dried on filter paper for analysis.

Statistical Analysis.
Tumor multiplicity was evaluated using one-way ANOVA followed by Kruskal-Wallis multiple comparisons test. Preneoplastic lesions and blood sample analysis were compared with NMBA controls using one-way ANOVA followed by Newman-Keuls range test. Tumor incidence was compared using ² analysis. Tumor size data were analyzed by linear regression, one-way ANOVA, Kruskal-Wallis multiple comparisons test, and the Dunnett test.

	RESULTS

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Toxicity.
Animals were fed normal diet or diet containing 0.5 or 1.0% POH beginning 3 days after carcinogen administration according to the treatment groups outlined in Table 1 . Animals in groups 3 and 5 that received 1.0% POH in their diets had a significant food aversion, consuming 5 g less diet/day than the other groups during the first week of exposure (Fig. 1) . This gradually improved but did not return to the consumption exhibited by groups 1, 2, and 4 until week 10. Groups 3 and 5 showed a concomitant decrease in body weight gain during the first weeks of 1.0% POH administration (Fig. 2) . At week 12, the rate of weight gain in these two groups was similar and remained so for the remainder of the study. However, the initial slowing of weight gain resulted in a >10% difference in body weight in groups 3 and 5 compared with control at the termination of the study. Group 4, receiving 0.5% POH mixed in their diets, had no aversion to food or decrease in weight gain.

View larger version (21K): [in this window] [in a new window] [Download PPT slide]   Fig. 1. Food consumption. Consumption was measured daily in grams and graphed over time. , group 1, DMSO control; , group 2, NMBA control; , group 3, POH control; x, group 4, 0.5% POH; *, group 5, 1.0% POH.

View larger version (22K): [in this window] [in a new window] [Download PPT slide]   Fig. 2. Body weights. Body weight was measured weekly and graphed over time. , group 1, DMSO control, , group 2, NMBA control; , group 3, POH control; x, group 4, 0.5% POH; *, group 5, 1.0% POH.

Tumor Data.
Esophageal tumors were counted and measured immediately after euthanization of rats. Animals treated with vehicle control or 1.0% POH alone did not have tumors. There were no significant differences in tumor incidence between groups 2 (77.4%), 4 (83.3%), and 5 (80.6%; Table 2 ). Analysis of tumor multiplicity in animals treated with NMBA plus 0.5% POH (2.09 tumors/rat) and 1.0% POH (2.08 tumors/rat) as compared with the NMBA control (1.54 tumors/rat) revealed a trend toward increasing tumors/rat in POH-fed animals. However, this was not statistically significant. Kruskal-Wallis multiple comparisons test results in z-values of 1.59 and 1.62 for the 0.5 and 1.0% POH-treated groups, respectively. This trend toward an increase in tumor multiplicity in POH-treated animals is not dose related.

Preneoplastic Lesions.
H&E-stained esophageal tissues were evaluated for preneoplastic changes. Slides were examined at x200 magnification, and each field was classified as normal, hyperplastic, and dysplastic. A statistically significant increase in dysplastic lesions was seen in both POH-treated groups as compared with NMBA control (Table 3 ; P < 0.05).

View larger version (31K): [in this window] [in a new window] [Download PPT slide]   Fig. 3. Representative Western blot. Membrane and cytosolic protein was extracted from esophagi of rats treated with NMBA alone (group 2) or NMBA + 1.0% POH (group 5) and subject to Western blot analysis with pan-ras antibody. Lanes 1, 2, and 4, SW480-positive control. Lanes 3 and 5: group 5 membrane proteins. Lanes 6 and 8, group 5 cytosolic proteins. Lanes 7 and 9, group 2 membrane proteins. Lane 10, group 2 cytosolic proteins. Protein standard.

	DISCUSSION

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This lack of chemopreventive efficacy in the esophagus may have several explanations. We have previously shown that the large majority of papillomas in our model contain Ha-ras GA transition mutations (34 , 37) . However, our data show that Ras membrane association was not affected by 1.0% POH administration in the diet. This is consistent with at least one report in the literature demonstrating that monoterpenes did not affect Ras cellular distribution in viral Ha-ras-transformed rat liver epithelial cells (21) . Similarly, a human Phase I trial indicates that in breast and prostate cancer cells, Ras expression and isoprenylation status was not affected at plasma concentrations of POH achieved by nontoxic doses (11) . Therefore, it is unlikely that POH is effectively inhibiting Ras signaling in this study.

An additional mechanism by which POH is believed to act involves the induction of apoptosis through the TGF-ß pathway. TGF-ß1 is secreted as inactive complex and must be released from this complex to become biologically active (38) . This latent complex has been shown to bind to the M6P/IGF-IIR (39) , and this binding facilitates the proteolytic activation of TGF-ß (40) . Up-regulation of the M6P/IGF-IIR has been seen in animal studies with POH and limonene, resulting in cytostasis and apoptosis (4 , 8 , 24) . Increased expression of M6P/IGF-IIR and TGF-ß1 is associated with mammary tumor regression, and only tumors showing this response regress (8) . However, in the NMBA-induced rat model of esophageal carcinogenesis, expression of TGF-ß1 is not increased until late in tumor development, at 45 weeks (41) . Animals in this current study were sacrificed at 25 weeks. Therefore, an increase in M6P/IGF-IIR mediated by POH would not have maximal effect until late in papilloma development.

This is the second model system we are aware of in which POH appears to enhance early tumorigenesis. Low-Baselli et al. (42) recently showed a similar promoting effect in rat hepatocarcinogenesis, demonstrating an increase in preneoplastic G+ foci after treatment with escalating dose of POH (maximum dose 1.8%). This study hypothesized that the enhancement may be attributable to decreased apoptosis, as well as the ability of monoterpenes to inhibit intercellular communication. Previous work with rat esophageal epithelial cells shows altered heterologous gap-junctional intercellular communication in neoplastic cells (43) . Therefore, it is possible that this is a mechanism by which POH promotes. Additionally, the esophagus is among the first organs exposed to any compound administered in the diet. Although no evidence of toxicity was observed in the stomach, an increase in nuclear and cytoplasmic vacuolization was noted in all groups treated with POH during histopathological evaluation of the esophagus that may indicate nonspecific toxicity. It is possible that a local toxic effect resulted in increased cellular proliferation and an enhancement of postinitiation effects by POH.

We observed a statistically significant decrease in tumor size in the high-dose POH group. This did not result in a decrease in total tumor volume/rat, however, a nonsignificant trend toward decreasing tumor burden in the 1.0% POH-treated group was observed. This adds to the growing body of literature suggesting that monoterpenes have differential effects early and late in tumorigenesis (42 , 44) . It is possible that the decrease in tumor size is related to monoterpene’s demonstrated efficacy in regressing tumors (5, 6, 7, 8, 9) . However, this is confounded by the decreased dietary intake observed in this treatment group. It is well known that caloric restriction can inhibit tumorigenesis. Although rats were fed ad lib synthetic diet and were not restricted in the classic sense, previous work in the zinc-deficient model of NMBA-induced rat esophageal cancer (45 , 46) has demonstrated an association between decreased dietary intake and decreased cell proliferation. It is possible that the tumors were inhibited by food aversion alone. In fact, it is conceivable that decreased dietary intake may have negatively affected tumor multiplicity, masking an even greater enhancing effect of high-dose POH. Therefore, although we conclude that POH is unlikely to be effective as a chemopreventive agent in squamous cell carcinoma of the esophagus, we are unable to comment on its potential as a chemotherapeutic agent.

In summary, POH showed a slight enhancing effect early in NMBA-induced rat esophageal tumorigenesis. In the high-dose group, a slight decrease in tumor size was noted, possibly indicating an effect on tumor growth or regression later in tumor development. High dietary concentrations of POH did not affect Ras membrane localization and may not be a feasible target of action for this agent. Our data suggest caution for the use of POH as a chemopreventive agent for squamous cell carcinoma of the esophagus.

	ACKNOWLEDGMENTS

 
We thank Dr. Thomas Knobloch for his help with the Western blots and Dr. Mark Morse for his aid in preparation of this manuscript. We also thank the Ohio State University School of Public Health Biostatistics Program for help in conducting the data analysis.

	FOOTNOTES

 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work supported by National Cancer Institute Grant PO1 CA46535.

² To whom requests for reprints should be addressed, at The Ohio State University, School of Public Health, CHRI, Room 1148, 300 West 10^th Avenue, Columbus, OH 43210. Phone: (614) 293-3713; Fax: (614) 293-3333.

³ The abbreviations used are: POH, perillyl alcohol; NMBA, N-nitrosomethylbenzylamine; TGF-ß, transforming growth factor ß; M6P, mannose-6-phosphate; IGF-IIR, insulin-like growth factor II receptor.

Received 8/19/02. Accepted 3/18/03.

	REFERENCES

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 

1. Lantry L. E., Zhang Z., Gao F., Crist K. A., Wang Y., Kelloff G. J., Lubet R. A., You M. Chemopreventive effect of perillyl alcohol on 4-(methylnitrosamine)-1-(3-pyridyl)-1-butanone induced tumorigenesis in (C3H/HeJ X A/J) F1 mouse. J. Cell. Biochem. Suppl., 27: 20-25, 1997.[Medline]
2. Barthelman M., Chen W., Gensler H. L., Huang C., Dong Z., Bowden G. T. Inhibitory effects of perillyl alcohol on UVB-induced murine skin cancer and AP-1 transactivation. Cancer Res., 58: 711-716, 1998.[Abstract/Free Full Text]
3. Reddy B. S., Wang C. X., Samaha H., Lubet R., Steele V. E., Kelloff G. J., Rao C. V. Chemoprevention of colon carcinogenesis by dietary perillyl alcohol. Cancer Res., 57: 420-425, 1997.[Abstract/Free Full Text]
4. Mills J. J., Chari R. S., Boyer I. J., Gould M. N., Jirtle R. L. Induction of apoptosis in liver tumors by the monoterpene perillyl alcohol. Cancer Res., 55: 979-983, 1995.[Abstract/Free Full Text]
5. Haag J. D., Gould M. N. Mammary carcinoma regression induced by perillyl alcohol, a hydroxylated analog of limonene. Cancer Chemother. Pharmacol., 34: 477-483, 1994.[Medline]
6. Elegbede J. A., Elson C. E., Tanner M. A., Qureshi A., Gould M. N. Regression of rat primary mammary tumors following dietary d-limonene. J. Natl. Cancer Inst. (Bethesda), 76: 323-325, 1986.
7. Haag J. D., Lindstrom M. J., Gould M. N. Limonene-induced regression of mammary carcinomas. Cancer Res., 52: 4021-4026, 1992.[Abstract/Free Full Text]
8. Jirtle R. L., Haag J. D., Ariazi E. A., Gould M. N. Increased mannose 6-phosphate/insulin-like growth factor II receptor and transforming growth factor ß 1 levels during monoterpene-induced regression of mammary tumors. Cancer Res., 53: 3849-3852, 1993.[Abstract/Free Full Text]
9. Stark M. J., Burke Y. D., McKinzie J. H., Ayoubi A. S., Crowell P. L. Chemotherapy of pancreatic cancer with the monterpene perillyl alcohol. Cancer Lett., 96: 15-21, 1995.[Medline]
10. Ripple G. H., Gould M. N., Arzoomanian R. Z., Alberti D., Feierabend C., Simon K., Binger K., Tutsch K. D., Pomplun M., Wahamaki A., Marnocha R., Wilding G., Bailey H. H. Phase I clinical and pharmacokinetic study of perillyl alcohol administered four times a day. Clin. Cancer Res., 6: 390-396, 2000.[Abstract/Free Full Text]
11. Hudes G. R., Sazrka C. E., Adams A., Ranganathan S., McCauley R. A., Weiner L. M., Langer C. J., Litwin S., Yeslow G., Halberr T., Qian M., Gallo J. M. Phase I pharmacokinetic trial of perillyl alcohol (NSC 641066) in patients with refractory solid malignancies. Clin. Cancer Res., 6: 3071-3080, 2000.[Abstract/Free Full Text]
12. Crowell P. L., Ren Z., Lin S., Vedejz E., Gould M. N. Structure-activity relationships among monoterpene inhibitors of protein isoprenylation and cell proliferation. Biochem. Pharmacol., 47: 1405-1415, 1994.[Medline]
13. Crowell P. L., Chang R. R., Ren Z. B., Elson C. E., Gould M. N. Selective inhibition of isoprenylation of 21–26-kDa proteins by the anticarcinogen d-limonene and its metabolites. J. Biol. Chem., 266: 17679-17685, 1991.[Abstract/Free Full Text]
14. Gelb M. H., Tamonoi F., Yokoyama K., Ghomashchi F., Esson K., Gould M. N. The inhibition of protein prenyltransferases by oxygenated metabolites of limonene and perillyl alcohol. Cancer Lett., 91: 169-175, 1995.[Medline]
15. Schulz S., Buhling F., Ansorge S. Prenylated proteins and lymphocyte proliferation: inhibition by d-limonene related monoterpenes. Eur. J. Immunol., 24: 301-307, 1994.[Medline]
16. Kato K., Cox A. D., Hisaka M. M., Graham S. M., Buss J. E., Der C. J. Isoprenoid addition to Ras protein is the critical modification for its membrane association and transforming activity. Proc. Natl. Acad. Sci. USA, 89: 6403-6407, 1992.[Abstract/Free Full Text]
17. Hardcastle I. R., Rowlands M. G., Barber A. M., Grimshaw R. M., Mohan M. K., Nutley B. P., Jarman M. Inhibition of protein prenylation by metabolites of limonene. Biochem. Pharmacol., 57: 801-809, 1999.[Medline]
18. Crowell P. L., Lin S., Vedejs E., Gould M. N. Identification of metabolites of the antitumor agent d-limonene capable of inhibiting protein isoprenylation and cell growth. Cancer Chemother. Pharmacol., 31: 205-212, 1992.[Medline]
19. Phillips L. R., Malspeis L., Supko J. G. Pharmacokinetics of active drug metabolites after oral administration of perillyl alcohol, an investigational antineoplastic agent, to the dog. Drug Metab. Dispos., 23: 676-680, 1995.[Abstract]
20. Ren Z., Gould M. N. Modulation of small G protein isoprenylation by anticancer monoterpenes in in situ mammary gland epithelial cells. Carcinogenesis (Lond.), 19: 827-832, 1998.[Abstract/Free Full Text]
21. Ruch R. J., Sigler K. Growth inhibition of rat liver epithelial tumor cells by monoterpenes does not involve Ras plasma membrane association. Carcinogenesis (Lond.), 15: 787-789, 1994.[Abstract/Free Full Text]
22. Shi W., Gould M. N. Induction of cytostasis in mammary carcinoma cells treated with the anticancer agent perillyl alcohol. Carcinogenesis (Lond.), 23: 131-142, 2002.[Abstract/Free Full Text]
23. Bardon S., Picard K., Martel P. Monoterpenes inhibit cell growth, cell cycle progression, and cyclin D1 gene expression in human breast cancer cell lines. Nutr. Cancer, 32: 1-7, 1998.[Medline]
24. Ariazi E. A., Satomi Y., Ellis M. J., Haag J. D., Shi W., Sattler C. A., Gould M. N. Activation of the transforming growth factor ß signaling pathway and induction of cytostasis and apoptosis in mammary carcinomas treated with the anticancer agent perillyl alcohol. Cancer Res., 59: 1917-1928, 1999.[Abstract/Free Full Text]
25. Stayrook K. R., McKinzie J. H., Burke Y. D., Burke Y. A., Crowell P. L. Induction of the apoptosis-promoting protein Bak by perillyl alcohol in pancreatic ductal adenocarcinoma relative to untransformed ductal epithelial cells. Carcinogenesis (Lond.), 18: 1655-1658, 1997.[Abstract/Free Full Text]
26. Wei X., Si M. S., Imagawa D. K., Ji P., Tromberg B. J., Cahalan M. D. Perillyl alcohol inhibits TCR-mediated Ca[2+] signaling, alters cell shape and motility and induced apoptosis in T lymphocytes. Cell. Immunol., 201: 6-13, 2000.[Medline]
27. Satomi Y., Miyamoto S., Gould M. N. Induction of AP-1 activity by perillyl alcohol in breast cancer cells. Carcinogenesis (Lond.), 20: 1957-1961, 1999.[Abstract/Free Full Text]
28. American Institute for Cancer Res. Food, nutrition and the prevention of cancer. World Cancer Res. Fund, : 188-229, 1997.
29. Yang C. Research in esophageal cancer in China: A review. Cancer Res., 40: 2633-2644, 1980.[Abstract/Free Full Text]
30. Stoner G. D., Rustgi A. K. Biology of esophageal squamous cell carcinoma Rustgi A. K. eds. . Gastrointestinal cancers: biology, diagnosis and therapy, 141-147, Lippincott-Raven Philadelphia 1995.
31. Druckery H. Organospecific carcinogenesis in the digestive tract Odashima S. eds. . Topics in Chemical Carcinogenesis, 73-101, University Park Press Baltimore 1972.
32. Lijinsky W., Rueber M. D. Carcinogenicity in rats of nitrosomethylethylamines labeled with deuterium in several positions. Cancer Res., 40: 19-21, 1980.[Abstract/Free Full Text]
33. Lijinsky W., Saavedra J. E., Reuber M. D., Singer S. S. Esophageal carcinogenesis in F344 rats by nitrosomethylethylamines substituted in the ethyl group. J. Natl. Cancer Inst. (Bethesda), 68: 681-684, 1982.
34. Wang Y., You M., Reynolds S. H., Stoner G. D., Anderson M. W. Mutational activation of the cellular Harvey ras oncogene in rat esophageal papillomas induced by methylbenzylnitrosamine. Cancer Res., 50: 1591-1595, 1990.[Abstract/Free Full Text]
35. Barch D. H., Jacoby R. F., Brasitus T. A., Radosevich J. A., Carney W. P., Iannaccone P. M. Incidence of Harvey ras oncogene point mutations and their expression in methylbenzylnitrosamine-induced esophageal tumorigenesis. Carcinogenesis (Lond.), 12: 2373-2377, 1991.[Abstract/Free Full Text]
36. Lozano J. C., Nakazawa H., Cros M. P., Cabral R., Yamasaki H. GA mutation in p53 and Ha-ras genes in esophageal papillomas induced by N-nitrosomethylbenzylamine in two strains of rats. Mol. Carcinog., 9: 33-39, 1994.[Medline]
37. Liston B. W., Gupta A., Nines R., Carlton P. S., Kresty L. A., Harris G. K., Stoner G. D. Incidence and effects of Ha-ras codon 12 GA transition mutations in preneoplastic lesions induced by N-nitrosomethylbenzylamine in the rat esophagus. Mol. Carcinog., 32: 1-8, 2001.[Medline]
38. Lyons R. M., Keski-Oja J., Moses H. L. Proteolytic activation of latent transforming growth factor ß from fibroblast-conditioned medium. J. Cell. Biol., 106: 1659-1665, 1988.[Abstract/Free Full Text]
39. Kornfeld S. Structure and function of the mannose 6-phosphate/insulin-like growth factor II receptor. Annu. Rev. Biochem., 61: 307-330, 1992.[Medline]
40. Dennis P. A., Rifkin D. B. Cellular activation of latent transforming growth factor ß requires binding to the cation-independent mannose 6-phosphate/insulin-like growth factor type II receptor. Proc. Natl. Acad. Sci. USA, 88: 580-584, 1991.[Abstract/Free Full Text]
41. Wang Q-S., Sabourin C. L., Kresty L. A., Stoner G. D. Dysregulation of transforming growth factor ß 1 expression in N-nitrosomethylbenzylamine-induced rat esophageal tumorigenesis. Int. J. Oncol., 9: 473-479, 1996.
42. Low-Baselli A., Huber W. W., Kafer M., Bukowska K., Schulte-Hermann R., Grasl-Kraupp B. Failure to demonstrate chemoprevention by the monoterpene perillyl alcohol during early rat hepatocarcinogenesis: a cautionary note. Carcinogenesis (Lond.), 21: 1869-1877, 2002.
43. Garber S. A., Fernstrom M. J., Stoner G. D., Ruch R. J. Altered gap junctional intercellular communication in neoplastic rat esophageal epithelial cells. Carcinogenesis (Lond.), 18: 1149-1153, 1997.[Abstract/Free Full Text]
44. Kimura J., Takahashi S., Ogiso T., Yoshida Y., Akagi K., Hasegawa R., Kurata M., Hirose M., Shirai T. Lack of chemoprevention effects of the monoterpene d-limonene in a rat multi-organ carcinogenesis model. Jpn. J. Cancer Res., 87: 589-594, 1996.[Medline]
45. Fong L. Y. Y., Li J. X., Farber J. L., Magee P. N. Cell proliferation and esophageal carcinogenesis in the zinc-deficient rat. Carcinogenesis (Lond.), 17: 1841-1848, 1996.[Abstract/Free Full Text]
46. Fong L. Y. Y., Farber J. L., Magee P. N. Zinc replenishment reduces esophageal cell proliferation and N-nitrosomethylbenyzlamine (NMBA)-induced esophageal tumor incidence in zinc-deficient rats. Carcinogenesis (Lond.), 19: 1501-1596, 1998.

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