RESEARCH PAPER
D-limonene possesses cytotoxicity to tumor cells but not to hepatocytes
 
More details
Hide details
1
Department of Pharmacology, LM College of Pharmacy, Ahmedabad, India
 
2
Departments of Pharmacology and Toxicology, BV Patel Pharmaceutical Education and Research Development Centre, Ahmedabad, India
 
 
Submission date: 2017-10-06
 
 
Acceptance date: 2018-12-13
 
 
Online publication date: 2018-06-16
 
 
Corresponding author
Anita Mehta   

Department of Pharmacology, LM College of Pharmacy, Navrangpura, Ahmedabad-380009, India. Tel.: +91 9428418611.
 
 
Pol. Ann. Med. 2019;26(2):98-104
 
KEYWORDS
ABSTRACT
Introduction:
K562, human chronic myeloid leukemia cell line shows presence of constitutively active BCR–ABL gene. D-limonene, monoterpenes obtained from essential oils of citrus fruits have exhibited its antitumor activity in various types of cancers. Murine xenograft models are used for estimation of in vivo effect of D-limonene in K562 tumor xenograft model.

Aim:
The aim of present study was to evaluate the in vitro and in vivo effect of D-limonene on primary hepatocytes and K562 tumor implanted C57BL/6 mice respectively.

Material and methods:
Effect of D-limonene on growth of K562 cells and mouse primary hepatocytes was determined in vitro by MTT assay. The in vivo effect of D-limonene was also determined on chemically immunocompromised K562 tumor xenografted C57BL/6 mice.

Results and discussion:
In vitro dose dependent and time dependent studies of D-limonene shows significant reduction in viability of K562 cells. Dose dependent studies of doxorubicin and D-limonene treatment for 48 h shows significant reduction in viability of primary hepatocytes with doxorubicin whereas, the reduction was non significant with D-limonene. D-limonene treatment for 14 days also shows dose dependent reduction in tumor volume in K562 tumor xenograft C57BL/6 mice.

Conclusions:
D-limonene inhibited the growth of primary hepatocytes in-vitro and also inhibited in vivo K562 tumor growth in C57BL/6 mice, which suggests safety and efficacy of D-limonene in the treatment of chronic myeloid leukemia.

ACKNOWLEDGEMENTS
We express our deep sense of gratitude to B. V. Patel Pharmaceutical Education and Research Development (PERD) Centre for providing facilities and encouragement to conduct the research work. The financial assistance from DST-INSPIRE for junior research fellowship to Ms. Bhavini Shah (IF 140131) is gratefully acknowledged.
CONFLICT OF INTEREST
None.
 
REFERENCES (34)
1.
Deininger MWN, Goldman JM, Melo JV. The molecular biology of chronic myeloid leukemia. Blood. 2000;96(10):3343–3356.
 
2.
Goldman JM, Melo JV. Chronic myeloid leukemia – advances in biology and new approaches to treatment. N Engl J Med. 2003;349(15):1451–1464. https://doi.org/10.1056/NEJMra....
 
3.
Hochhaus A, O’Brien SG, Guilhot F, et al. Six-year follow-up of patients receiving imatinib for the first-line treatment of chronic myeloid leukemia. Leukemia. 2009;23:1054–1061. https://doi.org/10.1038/leu.20....
 
4.
O’Brien SG, Guilhot F, Larson RA, et al. Imatinib Compared with Interferon and Low-Dose Cytarabine for Newly Diagnosed Chronic-Phase Chronic Myeloid Leukemia. N Engl J Med. 2003;348:994–1004. https://doi.org/10.1056/NEJMoa....
 
5.
Kurzrock R, Kantarjian HM, Druker BJ, Talpaz M. Philadelphia chromosome-positive leukemias: From basic mechanisms to molecular therapeutics. Ann Intern Med. 2003;138(10):81–830. https://doi.org/10.7326/0003-4....
 
6.
Cortes J, Jabbour E, Kantarjian H, et al. Dynamics of BCR-ABL kinase domain mutations in chronic myeloid leukemia after sequential treatment with multiple tyrosine kinase inhibitors. Blood. 2007;110(12):4005–4011. https://doi.org/10.1182/blood-....
 
7.
Bixby D, Talpaz M. Mechanisms of resistance to tyrosine kinase inhibitors in chronic myeloid leukemia and recent therapeutic strategies to overcome resistance. ASH Education Program Book. 2009;2009(1):461–476. https://doi.org/10.1182/ashedu....
 
8.
Shah NP, Nicoll JM, Nagar B, et al. Multiple BCR-ABL kinase domain mutations confer polyclonal resistance to the tyrosine kinase inhibitor imatinib (STI571) in chronic phase and blast crisis chronic myeloid leukemia. Cancer Cell. 2002;2(2):117–125. https://doi.org/10.1016/S1535-....
 
9.
Cooper S, Giles FJ, Savona MR. Overcoming resistance in chronic myelogenous leukemia. Leuk Lymphoma. 2009;50(11):1785–1793. https://doi.org/10.3109/104281....
 
10.
Walz C, Sattler M. Novel targeted therapies to overcome imatinib mesylate resistance in chronic myeloid leukemia (CML). Crit. Rev. Oncol. Hematol. 2006;57(2):145–164. https://doi.org/10.1016/j.crit....
 
11.
Gould MN. Cancer chemoprevention and therapy by monoterpenes. Environ. Health Perspect. 1997;105(Suppl 4):977–979. https://doi.org/10.1289/ehp.97....
 
12.
Rabi T, Bishayee A. Terpenoids and breast cancer chemoprevention. Breast Cancer Res. Treat. 2009;115(2):223–239. https://doi.org/10.1007/s10549....
 
13.
Crowell PL, Gould MN. Chemoprevention and Therapy of Cancer by d-Limonene. Crit Rev Oncog. 1994;5(1):1–22. https://doi.org/10.1615/CritRe....
 
14.
Gao D, Xiao Z, Lü A-E. [Proliferation inhibition and apoptosis induction of K562 cells by D-limonene]. Zhongguo shi yan xue ye xue za zhi. 2006;14(6):1120–1122 [in Chinese].
 
15.
Guo X-M, Lu Q, Liu Z-J, Wang L-F, Feng B-A. [Effects of D-limonene on leukemia cells HL-60 and K562 in vitro]. Zhongguo shi yan xue ye xue za zhi. 2006;14(4):692–695 [in Cinese].
 
16.
Crowell PL, Ayoubi AS, Burke YD. Antitumorigenic Effects of Limonene and Perillyl Alcohol Against Pancreatic and Breast Cancer. Dietary Phytochemicals in Cancer Prevention and Treatment. Boston, MA: Springer US; 1996:131–136.
 
17.
Nakaizumi A, Baba M, Uehara H, Iishi H, Tatsuta M. D-Limonene inhibits N-nitrosobis(2-oxopropyl)amine induced hamster pancreatic carcinogenesis. Cancer Lett. 1997;117(1):99–103. https://doi.org/10.1016/S0304-....
 
18.
Miller JA, Thompson PA, Hakim IA, Chow H-HS, Thomson CA. d-Limonene: a bioactive food component from citrus and evidence for a potential role in breast cancer prevention and treatment. Oncol Rev. 2011;5(1):31–42. https://doi.org/10.4081/oncol.....
 
19.
Haag JD, Lindstrom MJ, Gould MN. Limonene-induced Regression of Mammary Carcinomas. Cancer Res. 1992;52(14):4021–4026.
 
20.
Lu XG, Zhan LB, Feng BA, Qu MY, Yu LH, Xie JH. Inhibition of growth and metastasis of human gastric cancer implanted in nude mice by d-limonene. World J Gastroenterol. 2004;10(14):2140–2144. https://doi.org/10.3748/wjg.v1....
 
21.
Guguen-Guillouzo C. Isolation and Culture of Animal and Human Hepatocytes. In: Freshney RI, Freshney MG, eds. Culture of Epithelial Cells. New York: Wiley & Sons; 2002:337–379. https://doi.org/10.1002/047122....
 
22.
Klaunig JE, Goldblatt PJ, Hinton DE, Lipsky MM, Chacko J, Trump BF. Mouse liver cell culture. I. Hepatocyte isolation. In Vitro. 1981;17(10):913–925. https://doi.org/10.1007/BF0261....
 
23.
Williams GM, Bermudez E, Scaramuzzino D. Rat hepatocyte primary cell cultures. In Vitro. 1977;13(12):809–817. https://doi.org/10.1007/BF0261....
 
24.
Jivrajani M, Shaikh MV, Shrivastava N, Nivsarkar M. An improved and versatile immunosuppression protocol for the development of tumor xenograft in mice. Anticancer Res. 2014;34(12):7177–7183.
 
25.
Carlsson G, Gullberg B, Hafstrom L. Estimation of liver tumor volume using different formulas - an experimental study in rats. J Cancer Res Clin Oncol. 1983;105(1):20–23. https://doi.org/10.1007/BF0039....
 
26.
Elson CE, Maltzman TH, Boston JL, Tanner MA, Gould MN. Anti-carcinogenic activity of d-limonene during the initiation and promotion/progression stages of DMBA-induced rat mammary carcinogenesis. Carcinogenesis. 1988;9(2):331–332. https://doi.org/10.1093/carcin....
 
27.
Maltzman TH, Hurt LM, Elson CE, Tanner MA, Gould MN. The prevention of nitrosomethylurea-induced mammary tumors by d-limonene and orange oil. Carcinogenesis. 1989;10(4):781–783. https://doi.org/10.1093/carcin....
 
28.
Vigushin DM, Poon GK, Boddy A, et al. Phase I and pharmacokinetic study of d-limonene in patients with advanced cancer. Cancer Chemother Pharmacol. 1998;42(2):111–117. https://doi.org/10.1007/s00280....
 
29.
Rabi T, Bishayee A. D -Limonene sensitizes docetaxel-induced cytotoxicity in human prostate cancer cells: Generation of reactive oxygen species and induction of apoptosis. J Carcinog. 2009;8:9–9. https://doi.org/10.4103/1477-3....
 
30.
Yang H, Ping Dou Q. Targeting Apoptosis Pathway with Natural Terpenoids: Implications for Treatment of Breast and Prostate Cancer. Curr Drug Targets. 2010;11(6):733–744. https://doi.org/10.2174/138945....
 
31.
Simunek T, Sterba M, Popelova O, Adamcova M, Hrdina R, Gersl V. Anthracycline-induced cardiotoxicity: overview of studies examining the roles of oxidative stress and free cellular iron. Pharmacol Rep. 2009;61(1):154–171. https://doi.org/10.1016/S1734-....
 
32.
Ng R, Better N, Green MD. Anticancer Agents and Cardiotoxicity. Semin Oncol. 2006;33(1):2–14. https://doi.org/10.1053/j.semi....
 
33.
Kalender Y, Yel M, Kalender S. Doxorubicin hepatotoxicity and hepatic free radical metabolism in rats: The effects of vitamin E and catechin. Toxicology. 2005;209(1):39–45. https://doi.org/10.1016/j.tox.....
 
34.
Roberto D, Micucci P, Sebastian T, Graciela F, Anesini C. Antioxidant activity of limonene on normal murine lymphocytes: relation to H2O2 modulation and cell proliferation. Basic Clin Pharmacol Toxicol. 2009;106(1):38–44. https://doi.org/10.1111/j.1742....
 
Journals System - logo
Scroll to top