Abstract
Background: Colon cancer is one of the most common cancers in both men and women. The present study is an effort to unravel the anticarcinogenic effects of rosmarinic acid (RA) in 1,2-dimethylhydrazine (DMH)-induced rat colon carcinogenesis. Administration of DMH induces multiple tumors in the rat colon, which mimics human colon cancer.
Methods: Male Wistar rats were divided into six groups and fed a high-fat diet. Group 1 served as control, group 2 rats were given RA [5 mg/kg body weight (b.w.)] orally every day for a total period of 30 weeks, and groups 3–6 were given weekly injections of DMH (20 mg/kg b.w. subcutaneous) once a week in the groin for the first 15 weeks. In addition to DMH, groups 4–6 received RA at a dose of 5 mg/kg b.w. during the initiation and postinitiation stages, and also throughout the entire study period. Colon tissues were examined histologically; further, the extent of oxidative stress was assessed by measuring lipid peroxidation and antioxidant levels in the colonic mucosa of rats.
Results: Macroscopic and microscopic tumors were identified in all the groups that received DMH. The results revealed that supplementation with RA significantly inhibited the tumor formation and tumor multiplicity in DMH-treated rats. RA supplementation to DMH-administered rats significantly reduced the cell proliferation markers, namely, argyrophilic nucleolar organizing regions as well as proliferative cell nuclear antigen labeling index. In addition, RA supplementation reduces the expressions of tumor necrosis factor-α, interlukin-6, and cyclooxygenase-2, and modulates the expression of p65.
Conclusions: The above findings clearly underline the chemopreventive efficacy of RA against DMH-induced colon carcinogenesis.
Acknowledgments
The financial assistance offered by the Indian Council of Medical Research, New Delhi, India, in the form of Senior Research Fellowship to V. Karthikkumar is gratefully acknowledged.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Abe S, Ogura S, Knikane H, Suko N, Watanabe N, Nakajima I, et al. Nucleolar organizer regions in precancerous and cancerous lesions of bronchus. Cancer (Phila.) 1991;67:472–5.Search in Google Scholar
2. Greenlee RT, Murray T, Bolden S, Wingo PA. Cancer statistics. CA Cancer J Clin 2000;50:7–33.10.3322/canjclin.50.1.7Search in Google Scholar
3. Johnson JJ, Mukhtar H. Curcumin for chemoprevention of colon cancer. Cancer Lett 2007;255:170–81.10.1016/j.canlet.2007.03.005Search in Google Scholar
4. Karthik kumar V, Vennila S, Nalini N. Modifying effects of morin on the development of aberrant crypt foci and bacterial enzymes in experimental colon cancer. Food Chem Toxicol 2009;47:309–15.10.1016/j.fct.2008.11.017Search in Google Scholar
5. Sangeetha N, Aranganathan S, Panneerselvam J, Shanthi P, Rama G, Nalini N. Oral supplementation of silibinin prevents colon carcinogenesis in a long term preclinical model. Eur J Pharmacol 2010;643:93–100.10.1016/j.ejphar.2010.05.060Search in Google Scholar
6. Fiala ES. Investigations into the metabolism and mode of action of the colon carcinogens 1,2-dimethylhydrazine and azoxymethane. Cancer 1977;40:2436–45.10.1002/1097-0142(197711)40:5+<2436::AID-CNCR2820400908>3.0.CO;2-USearch in Google Scholar
7. Feinberg A, Zedeck MS. Production of a highly reactive alkylating agent from the organospecific carcinogen methylazoxymethanol by alcohol dehydrogenase. Cancer Res 1980;40:4446–50.Search in Google Scholar
8. Wolter S, Frank N. Metabolism of 1,2-dimethylhydrazine in isolated perfused rat liver. Chem Biol Interact 1982;42:335–44.10.1016/0009-2797(82)90077-1Search in Google Scholar
9. Sugimura T. Multistep carcinogenesis: a perspective. Science 1992;258:603–7.10.1126/science.1411570Search in Google Scholar
10. Hanahan D, Weinberg RA. The hallmarks of cancer. Cell 2000;100:57–70.10.1016/S0092-8674(00)81683-9Search in Google Scholar
11. Vincent TL, Gatenby RA. An evolutionary model for initiation, promotion, and progression in carcinogenesis. Int J Oncol 2008;32:729–37.Search in Google Scholar
12. Miller AB, Bartsch H, Boffetta P, Dragsted L, Vainio H, editors. Biomarkers in cancer chemoprevention. Lyon: International Agency for Research on Cancer (IARC) Scientific Publication, 2001: 294pp.Search in Google Scholar
13. Smith R, Crocker J. Evaluation of nucleolar organizer region associated proteins in breast malignancy. Histopathology 1988;12:113–25.10.1111/j.1365-2559.1988.tb01923.xSearch in Google Scholar
14. Ruschoff J, Bittinger A, Neumann K, Schmitz-Moormann P. Prognostic significance of nucleolar organizing regions (NORs) in carcinomas of the sigmoid colon and rectum. Pathol Res Pract 1990;86:85–91.10.1016/S0344-0338(11)81014-9Search in Google Scholar
15. Ofner D, Tötsch M, Sandbichler P, Hallbrucker C, Margreiter R, Mikuz G, et al. Silver stained nucleolar organizer region proteins (Ag-NORs) as a predictor of prognosis in colonic cancer. J Pathol 1990;162:43–9.10.1002/path.1711620109Search in Google Scholar
16. Suarez V, Newman J, Hiley C, Crocker J, Collins M. The value of NOR numbers in neoplastic and non-neoplastic epithelium of stomach. Histopathology 1989;14:61–6.10.1111/j.1365-2559.1989.tb02114.xSearch in Google Scholar
17. Zhou DY, Feng FC, Zhang YL. Comparison of Shams’ test for rectal mucus to an immunological test for fecal occult blood in large intestinal carcinoma screening, analysis of a check-up of 6480 asymptomatic subjects. Chin Med J (Engl.) 1993;106:739–42.Search in Google Scholar
18. Demers RY, Stawick LE, Demers P. Relative sensitivity of the fecal occult blood test and flexible sigmoidoscopy in detecting polyps. Prev Med 1988;14:55–62.10.1016/0091-7435(85)90020-9Search in Google Scholar
19. Tsujii M, Kawano S, DuBois RN. Cyclooxygenase-2 expression in human colon cancer cells increases metastatic potential. Proc Natl Acad Sci USA 1997;94:3336–40.10.1073/pnas.94.7.3336Search in Google Scholar
20. Oeckinghaus A, Ghosh S. The NF-κB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol 2009;1:a000034.10.1101/cshperspect.a000034Search in Google Scholar
21. Gupta S, Bi R, Kim C, Chiplunkar S, Yel L, Gollapudi S. Role of NF-κB signaling pathway in increased tumor necrosis factor-alpha-induced apoptosis of lymphocytes in aged humans. Cell Death Differ 2005;12:177–83.10.1038/sj.cdd.4401557Search in Google Scholar
22. Rajasekaran A, Sivagnanam G, Xavier R. Nutraceuticals as therapeutic agents, a review research. J Pharm Tech 2008;1:328–40.Search in Google Scholar
23. del Bano MJ, Lorente J, Castillo J, Benavente-García O, del Río JA, Ortuño A, et al. Phenolic diterpenes, flavones, and rosmarinic acid distribution during the development of leaves, flowers, stems, and roots of Rosmarinus officinalis. Antioxidant activity. J Agric Food Chem 2003;51:4247–53.Search in Google Scholar
24. Kintzios S, Makri O, Panagiotopoulos E, Scapeti M. In vitro rosmarinic acid accumulation in sweet basil (Ocimum basilicum L.). Biotechnol Lett 2003;25:405–8.10.1023/A:1022402515263Search in Google Scholar
25. Osakabe N, Yasuda A, Natsume M, Sanbongi C, Kato Y, Osawa T, et al. Rosmarinic acid, a major polyphenolic component of Perilla frutescens, reduces lipopolysaccharide (LPS)-induced liver injury in D-galactosamine (D-GalN)-sensitized mice. Free Radic Biol Med 2002;33:798–6.10.1016/S0891-5849(02)00970-XSearch in Google Scholar
26. Osakabe N, Yasuda A, Natsume M, Yoshikawa T. Rosmarinic acid inhibits epidermal inflammatory responses, anticarcinogenic effect of Perilla frutescens extract in the murine two-stage skin model. Carcinogenesis 2004;25:549–57.10.1093/carcin/bgh034Search in Google Scholar PubMed
27. Kosar M, Goger F, Can Baser KH. In vitro antioxidant properties and phenolic composition of Salvia virgata Jacq. From Turkey. J Agric Food Chem 2008;56:2369–74.Search in Google Scholar
28. Huang S, Zheng R. Rosmarinic acid inhibits angiogenesis and its mechanism of action in vitro. Cancer Lett 2006;239:271–80.10.1016/j.canlet.2005.08.025Search in Google Scholar PubMed
29. McKay D, Blumberg J. A review of the bioactivity and potential health benefits of peppermint tea (Mentha piperita L.). Phytother Res 2006;20:619–33.10.1002/ptr.1936Search in Google Scholar
30. Venkatachalam K, Gunasekaran S, Jesudoss VA, Namasivayam N. The effect of rosmarinic acid on 1,2-dimethylhydrazine induced colon carcinogenesis. Exp Toxicol Pathol 2013;65:409–18.10.1016/j.etp.2011.12.005Search in Google Scholar
31. Bosman FT. Prognostic value of pathological characteristics of colorectal cancer. Eur J Cancer 2005;31A:1216–21.10.1016/0959-8049(95)00153-ASearch in Google Scholar
32. Nyska A, Zusman I, Klein T, Sheila N, Weis O, Madar Z, et al. Assessment of the nucleolar organizer regions by automated image analysis in benign and malignant colonic tumours and adjacent tissues in rats. J Comp Pathol 1995;113:45–50.10.1016/S0021-9975(05)80067-2Search in Google Scholar
33. Jiang ZY, Hunt JV, Wolff SP. Ferrous ion oxidation in the presence of xylenol orange for detection of lipid hydroperoxides in low density lipoprotein. Anal Biochem 1993;202:384.10.1016/0003-2697(92)90122-NSearch in Google Scholar
34. Kakkar PS, Das B, Viswanathan PN. A modified spectrophotometric assay for superoxide dismutase. Indian J Biochem Biophys 1984;21:130–2.Search in Google Scholar
35. Sinha KA. Colorimetric assay of catalase. Anal Biochem 1972;47:389–94.10.1016/0003-2697(72)90132-7Search in Google Scholar
36. Carlberg I, Mannervik B. Glutathione reductase. In: Meister A, editor. Methods in enzymology, vol. 7. New York: Academic Press, 1985:484.Search in Google Scholar
37. Rotruck JT, Pope AL, Ganther HE, Hafeman DG, Hoekstra WG. Selenium: biochemical role as a component of glutathione peroxidase. Science 1973;179:588–90.10.1126/science.179.4073.588Search in Google Scholar PubMed
38. Sinha R, Anderson DE, McDonald SS, Greenwald P. Cancer risk and diet in India. J Postgrad Med 2003;49:222–8.Search in Google Scholar
39. Moreno JJ. New aspects of the role of hydroxyeicosatetraenoic acids in cell growth and cancer development. Biochem Pharmacol 2009;77:1–10.10.1016/j.bcp.2008.07.033Search in Google Scholar PubMed
40. Das UN. Influence of polyunsaturated fatty acids and their metabolites on stem cell biology. Nutrition 2011;27:21–5.10.1016/j.nut.2010.04.003Search in Google Scholar PubMed
41. Serhan CN, Clish CB, Brannon J, Colgan SP, Chiang N, Gronert K. Novel functional sets of lipid-derived mediators with antiinflammatory actions generated from omega-3 fatty acids via cyclooxygenase 2-nonsteroidal antiinflammatory drugs and transcellular processing. J Exp Med 2000;192:1197–204.10.1084/jem.192.8.1197Search in Google Scholar PubMed PubMed Central
42. Arita M, Yoshida M, Hong S, Tjonahen E, Glickman JN, Petasis NA, et al. Resolvin E1, an endogenous lipid mediator derived from omega-3 eicosapentaenoic acid, protects against 2,4,6-trinitrobenzene sulfonic acid-induced colitis. Proc Natl Acad Sci USA 2005;102:7671–6.10.1073/pnas.0409271102Search in Google Scholar
43. Day DW, Jass JR, Price AB, Shepherd NA, Sloan Talbot IC, Warren BF, et al. Morson and Dawson’s gastrointestinal pathology, 4th ed. Berlin: Blackwell, 2003.10.1002/9780470755983Search in Google Scholar
44. Kawamori T, Tanaka T, Hara A, Yamahara J, Mori H. Modifying effects of naturally occurring products on the development of colonic aberrant crypt foci induced by azoxymethane in F344 rats. Cancer Res 1995;55:1277–82.Search in Google Scholar
45. Fakan S, Hernandez-Verdun D. The nucleolus and the nucleolar organizer regions. Biol Cell 1986;56:189–205.10.1111/j.1768-322X.1986.tb00452.xSearch in Google Scholar
46. Tanaka T, Takeuchi T, Nishikawa A, Takami T, Mori H. Nucleolar organizer regions in hepatocarcinogenesis induced by N-2-fluorenylacetamide in rats: comparison with bromodeoxyuridine immunohistochemistry. Jpn J Cancer Res 1989;80:1047–51.10.1111/j.1349-7006.1989.tb02257.xSearch in Google Scholar
47. Moldovan GL, Pfander B, Jentsch S. PCNA, the maestro of the replication fork. Cell 2007;129:665–79.10.1016/j.cell.2007.05.003Search in Google Scholar
48. Hur YG, Yun Y, Won J. Rosmarinic acid induces p56lck-dependent apoptosis in Jurkat and peripheral T cells via mitochondrial pathway independent from Fas/Fas ligand interaction. J Immunol 2004;172:79–87.10.4049/jimmunol.172.1.79Search in Google Scholar
49. Dianzani MU. Lipid peroxidation and cancer. Crit Rev Oncol Hematol 1993;15:125–47.10.1016/1040-8428(93)90052-6Search in Google Scholar
50. Sreedharan V, Venkatachalam KK, Namasivayam N. Effect of morin on tissue lipid peroxidation and antioxidant status in 1,2-dimethylhydrazine induced experimental colon carcinogenesis. Invest New Drugs 2009;27:21–30.10.1007/s10637-008-9136-1Search in Google Scholar
51. Tanaka T. Effect of diet on human carcinogenesis. Crit Rev Oncol Hematol 1997;25:73–95.10.1016/S1040-8428(96)00228-4Search in Google Scholar
52. Pillai MG, Thambi BS, Menon VP, Leelamma S. Influence of dietary fiber from coconut kernel (Cocus nucifera) on the 1,2-dimethylhydrazine induced lipid peroxidation in rats. J Nutr Biochem 1999;10:555–60.10.1016/S0955-2863(99)00035-2Search in Google Scholar
53. Matés JM, Pérez-Gómez C, Núñez de Castro I. Antioxidant enzymes and human diseases. Clin Biochem 1999;32:595–603.10.1016/S0009-9120(99)00075-2Search in Google Scholar
54. Rajeshkumar NV, Ramadasan K. Modulation of carcinogenic response and antioxidant enzymes of rats administered with 1,2-dimethylhydrazine by Picroliv. Cancer Lett 2003;191:137–43.10.1016/S0304-3835(02)00203-3Search in Google Scholar
55. Aranganathan S, Nalini N. Efficacy of the potential chemopreventive agent, hesperetin (citrus flavanone), on 1,2-dimethylhydrazine induced colon carcinogenesis. Food Chem Toxicol 2009;47:2594–600.10.1016/j.fct.2009.07.019Search in Google Scholar
56. Das UN. A radical approach to cancer. Med Sci Monit 2002;8:79–92.Search in Google Scholar
57. Cao H, Cheng W-X, Li C, Pan X-L, Xie X-G, Li T-H. DFT study on the antioxidant activity of rosmarinic acid. J Mol Struct 2005;719:177–83.10.1016/j.theochem.2005.01.029Search in Google Scholar
58. Huls G, Koornstra JJ, Kleibeuker JH. Non-steroidal anti-inflammatory drugs and molecular carcinogenesis of colorectal carcinomas. Lancet 2003;362:230–2.10.1016/S0140-6736(03)13915-3Search in Google Scholar
59. López-Posadas R, Ballester I, Abadía-Molina AC, Suárez MD, Zarzuelo A, Martínez-Augustin O, et al. Effect of flavonoids on rat splenocytes, a structure-activity relationship study. Biochem Pharmacol 2008;76:495–506.10.1016/j.bcp.2008.06.001Search in Google Scholar PubMed
60. Gurnell M, Wentworth JM, Agostini M, Adams M, Collingwood TN, Provenzano C, et al. A dominant-negative peroxisome proliferator-activated receptor γ (PPARγ) mutant is a constitutive repressor and inhibits PPARγ-mediated adipogenesis. J Biol Chem 2000;275:5754–9.10.1074/jbc.275.8.5754Search in Google Scholar PubMed
61. Konturek PC, Nikiforuk A, Kania J, Raithel M, Hahn EG, Muhldorfer S. Activation of NF-κB represents the central event in the neoplastic progression associated with Barrett’s esophagus: a possible link to the inflammation and overexpression of COX-2, PPARγ and growth factors. Dig Dis Sci 2004;49:1075–83.10.1023/B:DDAS.0000037790.11724.70Search in Google Scholar
62. Vaish V, Tanwar L, Sanyal SN. The role of NF-κB and PPARγ in experimentally induced colorectal cancer and chemoprevention by cyclooxygenase-2 inhibitors. Tumor Biol 2010;31:427–36.10.1007/s13277-010-0051-7Search in Google Scholar PubMed
63. Moon DO, Kim MO, Lee JD, Choi YH, Kim GY. Rosmarinic acid sensitizes cell death through suppression of TNF-α-induced NF-κB activation and ROS generation in human leukemia U937 cells. Cancer Lett 2010;288:183–91.10.1016/j.canlet.2009.06.033Search in Google Scholar PubMed
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