LOW LEVEL OF CYCLOOXYGENASE-2 TRANSCRIPT IN THE SPLEEN OF LYMPHOMA RATS SUPPLEMENTED WITH GARLIC POWDER

Authors

  • NURUL SAS
  • Hazilawati H Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia
  • SAJJARATTUL NNA
  • HUTHEYFA AH
  • ROSLY SM Animal Science Research Centre, Malaysian Agricultural Research and Development Institute, Serdang, Selangor, Malaysia
  • WAN SWZ Animal Science Research Centre, Malaysian Agricultural Research and Development Institute, Serdang, Selangor, Malaysia
  • NORHAIZAN ME Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor
  • SHANMUGAVELU S Animal Science Research Centre, Malaysian Agricultural Research and Development Institute, Serdang, Selangor, Malaysia
  • NOORDIN MM Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia
  • JASNI S Faculty of Veterinary Medicine, Universiti Malaysia Kelantan Kota Bharu, Kelantan, Malaysia

Abstract

Cyclooxygenases (COXs) regulate tumor cell proliferation and metastasis in many types of cancers including hematological cancers. Organosulfurs
derived from garlic have a potential to inhibit the expression of COX-2 in cancer patients.
Objective: In this study, we evaluate the transcription levels of COX-2 in the spleen of lymphoma rats supplemented with garlic powder.
Methods: Three groups of rats were equally divided into control (n=3), lymphoma (n=3), and lymphoma supplemented with garlic powder (n=3)
groups. Lymphoma was induced via administration of N-methyl-N-nitrosourea (MNU) intraperitoneally 4 times in 2 week periods. Garlic powder
mixed with ground commercial rat diet was given daily at 5% of feed intake, starting at day 1 of MNU exposure. All rats were kept for 24 weeks before
spleen samples were collected and extracted for total RNA. The transcription levels of COX-2 transcript in the total RNA were determined using
quantitative real-time reverse transcriptase polymerase chain reaction assay. The total RNA was converted into cDNA followed by amplification of
COX-2 and beta-actin genes.
Results: Results of the amplification of COX-2 transcripts were normalized with the housekeeping gene, beta-actin. The relative transcription level
of COX-2 transcript in the spleen of lymphoma rats was 1.941±0.131 fold higher (p<0.05) than control rats (1.00±0.001 fold), while the transcription
levels in the spleen of lymphoma rats supplemented with garlic was significantly lower (0.423±0.239 SE fold) than the lymphoma rats that received
no supplementation of garlic powder.
Conclusion: The findings suggest that garlic powder reduces the transcription of COX-2 transcript in the spleen of lymphoma rats
Keywords: Garlic, Cyclooxygenase-2, N-methyl-N-nitrosourea, Splenic lymphoma, Real-time reverse transcriptase polymerase chain reaction assay.

Downloads

Download data is not yet available.

References

World Health Organization (WHO). Report of the National Policy on

Traditional Medicine and Regulation of Herbal Medicine. Geneva:

World Health Organization; 2005.

Bernard S, Christopher PW. World Cancer Report 2014. Lyon:

International Agency for Cancer Research; 2014.

Kathiresan K, Boopathy NS, Kavitha S. Costal vegetation - An

underexplored source of anticancer drugs. Natl Prod Radiance

;5(2):115-9.

Ariga T Oshiba S. Platelet aggregation inhibitor in garlic. Lancet

;317:150-1.

Singh V, Belloir C, Siess MH, Le Bon AM. Inhibition of carcinogeninduced

DNA

damage in

rat liver and colon by garlic powders

with

varying

alliin content. Nutr Cancer 2006;55(2):178-84.

Boivin D, Simon LD, Jessica J, Edith B, Martine C, Albert M, et al.

Anti-proliferative and antioxidant activities of common vegetables:

A comparative study. Food Chem 2009;112(2):374-80.

Capasso A. Antioxidant action and therapeutic efficacy of Allium

sativum L. Molecules 2013;18(1):690-700.

Tao M, Gao L, Pan J, Wang X. Study on the inhibitory effect of allicin

on human gastric cancer cell line SGC-7901 and its mechanism. Afr J

Tradit Complement Altern Med 2013;11(1):176-9.

Secchiero P, Barbarotto E, Gonelli A, Tiribelli M, Zerbinati C,

Celeghini C, et al. Potential pathogenetic implications of

cyclooxygenase-2 overexpression in B chronic lymphoid leukemia

cells. Am J Pathol 2005;167(6):1599-607.

Barisik NO, Bozkurt S, Gumus M, Kaygusuz I, Karadayi N, Bas E,

et al. Expression and prognostic significance of Cox-2 and p-53 in

Hodgkin lymphomas: A retrospective study. Diagn Pathol 2010;5:19.

Katoh H, Hosono K, Ito Y, Suzuki T, Ogawa Y, Kubo H, et al. COX-2

and prostaglandin EP3/EP4 signaling regulate the tumor stromal

proangiogenic microenvironment via CXCL12-CXCR4 chemokine

systems. Am J Pathol 2010;176(3):1469-83.

Breyer RM, Bagdassarian CK, Myers SA, Breyer MD. Prostanoid

receptors: Subtypes and signalling. Ann Rev Pharmacol Toxicol

;41:661-90.

Stratton MS, Alberts DS. Current application of selective COX-2

inhibitors in cancer prevention and treatment. Oncology (Williston

Park) 2002;16 5 Suppl 4:37-51.

Bernard MP, Bancos S, Sime PJ, Phipps RP. Targeting cyclooxygenase-2

in hematological malignancies: Rationale and promise. Curr Pharm Des

;14(21):2051-60.

Asproni P, Vignoli M, Cancedda S, Millanta F, Terragni R, Poli A.

Immunohistochemical expression of cyclooxygenase-2 in normal,

hyperplastic and neoplastic canine lymphoid tissues. J Comp Pathol

;151(1):35-41.

Cao Y, Prescott SM. Many actions of cyclooxygenase-2 in cellular

dynamics and in cancer. J Cell Physiol 2002;190(3):279-86.

Chan MW, Wong CY, Cheng AS, Chan YW, Chan KK, To FK,

et al. Targeted inhibition of COX-2 expression by RNA interference

suppresses tumour growth and potentiates chemosensitivity to cisplatin

in human gastric cancer cells. Oncol Rep 2001;18:1557-62.

Liu B, Qu L, Yan S. Cyclooxygenase-2 promotes tumor growth and

suppresses tumor immunity. Cancer Cell Int 2015;15:106.

Gately S, Li WW. Multiple roles of COX-2 in tumor angiogenesis: A

target for antiangiogenic therapy. Semin Oncol 2004;31 2 Suppl 7:2-11.

Asian J Pharm Clin Res, Vol 9, Issue 3, 2016, 187-192

Nurul et al.

Le Bon AM, Vernevaut MF, Guenot L, Kahane R, Auger J, Arnault I,

et al. Effects of garlic powders with varying alliin contents on

hepatic drug metabolizing enzymes in rats. J Agric Food Chem

;51(26):7617-23.

Nie SN, Sun HC, Wu XH, Qian XM. Cyclooxygenase 2, pS2, inducible

nitric oxide synthase and transforming growth factor alpha in gastric

adaptation to stress. World J Gastroenterol 2004;10(23):3537-41.

Nursyuhada H, Hazilawati H, Hutheyfa AH, Rosly SM,

Shanmugavellu S, Noordin MM, et al. Detection of Bcl-2 in leukemic

rats using an Eva Green real-time RT-PCR assay. Pertanika J Trop Agric

Sci 2011;34:372-80.

Hutheyfa AH, Hazilawati H, Rosly SM, Jasni S, Noordin MM,

Shanmugavelu S. Histopathological features of peripheral T-cell

lymphoma in Sprague-Dawley rats induced with N-methyl-Nnitrosourea.

Pertanika J Trop

Agric

Sci 2011;34:357-68.

Maekawa A, Ogiu T, Onodera H, Furuta K, Matsuoka C, Ohno Y, et al.

Carcinogenicity studies of sodium nitrite and sodium nitrate in F-344

rats. Food Chem Toxicol 1982;20(1):25-33.

da Silva Franchi CA, Bacchi MM, Padovani CR, de Camargo JL.

Thymic lymphomas in Wistar rats exposed to N-methyl-N-nitrosourea

(MNU). Cancer Sci 2003;94(3):240-3.

Tsubura A, Lai LC, Miki Y, Tomo S, Norihisa U, Yuri T, et al. Animal

models of N-Methyl-N-nitrosourea-induced mammary cancer and

retinal degeneration with special emphasis on therapeutic trials. Int J

Exp Clin Pathophysiol Drug Res 2011;25(1):11-22.

Swann FP, Magee PN. Nitrosamine induced carcinogen. The

alkylation of nucleic acids of the rat by N-methyl-N-nitrosourea,

dimethylnitrosamine, dimethyl sulphate and methyl methanesulphate.

Biochem J 1968;110:39-47.

Chang YC, Hsu JD, Lin WL, Lee YJ, Wang CJ. High incidence of

acute promyelocytic leukemia specifically induced by N-nitrosoN-methylurea

(NMU)

in Sprague-Dawley

rats.

Arch

Toxicol

;86(2):315-27.

Sajjaratul NN, Hazilawati H, Rosly MS, Hakimi MK, Nursyuhada H,

Yusnaini MY, et al. Effect of different routes of N-methyl-N-nitrosourea

(MNU) exposure on the levels of vascular endothelial growth factor

(VEGF) transcript in spleen of rats. Malays J Vet Res 2014;5:184-5.

Gal AF, Miclăuș V, Taulescu M, Bolfă P, Tăbăran F, Bouari C, et al.

Concurrent mammary adenocarcinoma and lymphoma in MNUinduced

tumourigenesis

in a Sprague-Dawley female rat.

Bull UASMV

Vet

Med 2012;69:1-2.

Zeng T, Zhang CL, Zhao XL, Xie KQ. The roles of garlic on the lipid

parameters: A systematic review of the literature. Crit Rev Food Sci

Nutr 2013;53(3):215-30.

Neil A, Silagy C. Garlic: Its cardio-protective properties. Curr Opin

Lipidol 1994;5(1):6-10.

Rahman K. Historical perspective on garlic and cardiovascular disease.

J Nutr 2001;131(3s):977S-9.

Tanaka S, Haruma K, Yoshihara M, Kajiyama G, Kira K, Amagase H,

et al. Aged garlic extract has potential suppressive effect on colorectal

adenomas in humans. J Nutr 2006;136 3 Suppl:821S-6.

Zhou Y, Zhuang W, Hu W, Liu GJ, Wu TX, Wu XT. Consumption of

large amounts of Allium vegetables reduces risk for gastric cancer in a

meta-analysis. Gastroenterology 2011;141(1):80-9.

Ngo SN, Williams DB, Cobiac L, Head RJ. Does garlic reduce risk of

colorectal cancer? A systematic review. J Nutr 2007;137(10):2264-9.

Thomson M, Ali M. Garlic [Allium sativum]: A review of its potential

use as an anti-cancer agent. Curr Cancer Drug Targets 2003;3(1):67-81.

Mocellin S, Bronte V, Nitti D. Nitric oxide a double edged sword in

cancer biology: Searching for therapeutic opportunities. Med Res Rev

;27(3):317-52.

Wink DA, Hines HB, Cheng RY, Switzer CH, Flores-Santana W,

Vitek MP, et al. Nitric oxide and redox mechanisms in the immune

response. J Leukoc Biol 2011;89(6):873-91.

Wang HC, Yang JH, Hsieh SC, Sheen LY. Allyl sulfides inhibit cell

growth of skin cancer cells through induction of DNA damage mediated

G2/M arrest and apoptosis. J Agric Food Chem 2010;58(11):7096-103.

Schäfer G, Kaschula CH. The immunomodulation and antiinflammatory

effects

of garlic organosulfur

compounds in cancer

chemoprevention.

Anticancer

Agents

Med Chem 2014;14(2):233-40.

Lin WW, Karin M. A cytokine-mediated link between innate immunity,

inflammation, and cancer. J Clin Invest 2007;117(5):1175-83.

Kleinert H, Schwarz PM, Förstermann U. Regulation of the expression

of inducible nitric oxide synthase. J Biol Chem 2003;384:1343-64.

Massi D, Franchi A, Sardi I, Magnelli L, Paglierani M, Borgognoni L,

et al. Inducible nitric oxide synthase expression in benign and malignant

cutaneous melanocytic lesions. J Pathol 2001;194(2):194-200.

Wun T, McKnight H, Tuscano JM. Increased cyclooxygenase-2

(COX-2): A potential role in the pathogenesis of lymphoma. Leuk Res

;28(2):179-90.

Hazar B, Ergin M, Seyrek E, Erdogan S, Tuncer I, Hakverdi S.

Cyclooxygenase-2 (Cox-2) expression in lymphomas. Leuk Lymphoma

;45(7):1395-9.

Wang XH, Shao DH, Liang GW, Zhang R, Xin Q, Zhang T, et al.

Cyclooxygenase inhibitors in some dietary vegetables inhibit platelet

aggregation function induced by arachidonic acid. Zhongguo Shi Yan

Xue Ye Xue Za Zhi 2011;19(5):1260-3.

Archana S, Samit G, Shamee B, Sukta D. Experimental evaluation of

anticarcinogenic effect of garlic in rats colon cancer. Asian Pac J Cancer

;5:126-32.

Hong CH, Hur SK, Oh OJ, Kim SS, Nam KA, Lee SK. Evaluation of

natural products on inhibition of inducible cyclooxygenase (COX-2)

and nitric oxide synthase (iNOS) in cultured mouse macrophage cells.

J Ethnopharmacol 2002;83(1-2):153-9.

Lev-Ari S, Starr A, Vexler A, Karaush V, Loew V, Greif J, et al.

Inhibition of pancreatic and lung adenocarcinoma cell survival by

curcumin is associated with increased apoptosis, down-regulation of

COX-2 and EGFR and inhibition of Erk1/2 activity. Anticancer Res

;26(6B):4423-30.

Kwon HK, Jeon WK, Hwang JS, Lee CG, So JS, Park JA, et al.

Cinnamon extract suppresses tumour progression by modulating

angiogenesis and the effector function of CD8+ T cells. Cancer Lett

;278(2):174-82.

Brock TG, McNish RW, Peters-Golden M. Arachidonic acid is

preferentially metabolized by cyclooxygenase-2 to prostacyclin and

prostaglandin E2. J Biol Chem 1999;274(17):11660-6.

Harris RE. Cyclooxygenase-2 (COX-2) and the inflammogenesis of

cancer. Subcell Biochem 2007;42:93-126.

Li HJ, Reinhardt F, Herschman HR, Weinberg RA. Cancer-stimulated

mesenchymal stem cells create a carcinoma stem cell niche via

prostaglandin E2 signalling. Cancer Discov 2012;2(9):840-55.

Banning A, Florian S, Deubel S, Thalmann S, Müller-Schmehl K,

Jacobasch G, et al. GPx2 counteracts PGE2 production by dampening

COX-2 and mPGES-1 expression in human colon cancer cells. Antioxid

Redox Signal 2008;10(9):1491-500.

Soni K, Shashi PM, Shyam BP, Suresh SY, Mitali D, Mohan K, et al.

Differential expression of COX2: A potential marker for clinical

phenotypes of gastric cancer. Int J Adv Res 2014;2(2):278-88.

Konturek PC, Konturek SJ, Pierzchalski P, Starzyńska T,

Marlicz K, Hartwich A, et al. Gastric MALT-lymphoma, gastrin and

cyclooxygenases. Acta Gastro Enterol Belg 2002;65(1):17-23.

Kopp KL, Kauczok CS, Lauenborg B, Krejsgaard T, Eriksen KW,

Zhang Q, et al. COX-2-dependent PGE(2) acts as a growth factor in

mycosis fungoides (MF). Leukemia 2010;24(6):1179-85.

Massi DV, Alexandra KK, Hildebert W, Angelika MV. Effect of allicin

and ajoene, two compounds of garlic, on inducible nitric oxide synthase.

Atherosclerosis 1998;139(2):333-9.

Chang HP, Chen YH. Differential effects of organosulfur compounds

from garlic oil on nitric oxide and prostaglandin E2 in stimulated

macrophages. Nutrition 2005;21(4):530-6.

Lang A, Lahav M, Emad S, Iris B, Herma HF, Benjamin A, et al. Allicin

inhibits spontaneous and TNF-α induced secretion of proinflammatory

cytokines and chemokines from intestinal epithelial cells. Clin Nutr

;23(5):1199-208.

Oommen S, Anto RJ, Srinivas G, Karunagaran D. Allicin (from garlic)

induces caspase-mediated apoptosis in cancer cells. Eur J Pharmacol

;485(1-3):97-103.

Pari L, Murugavel P, Sitasawad SL, Kumar KS. Cytoprotective and

antioxidant role of diallyl tetrasulfide on cadmium induced renal injury:

An in vivo and in vitro study. Life Sci 2007;80(7):650-8.

Son EW, Mo SJ, Rhee DK, Pyo S. Inhibition of ICAM-1 expression

by garlic component, allicin, in gamma-irradiated human vascular

endothelial cells via downregulation of the JNK signalling pathway. Int

Immunopharmacol 2006;6(12):1788-95.

Murugavel P, Pari L. Effects of diallyl tetrasulfide on cadmiuminduced

oxidative

damage

in

the

liver

of rats. Hum

Exp Toxicol

;26(6):527-34.

Uefuji K, Ichikura T, Mochizuki H. Cyclooxygenase-2 expression is

related to prostaglandin biosynthesis and angiogenesis in human gastric

cancer. Clin Cancer Res 2000;6(1):135-8.

Uefuji K, Ichikura T, Mochizuki H. Expression of cyclooxygenase-2

in human gastric adenomas and adenocarcinomas. J Surg Oncol

;76(1):26-30.

Koh YW, Park C, Yoon DH, Suh C, Huh J. Prognostic significance of

Asian J Pharm Clin Res, Vol 9, Issue 3, 2016, 187-192

Nurul et al.

COX-2 expression and correlation with Bcl-2 and VEGF expression,

microvessel density, and clinical variables in classical Hodgkin

lymphoma. Am J Surg Pathol 2013;37(8):1242-51.

Tong Z, Gao Z, Wang F, Zhou J, Zhang Z. Selection of reliable reference

genes for gene expression studies in peach using real-time PCR. BMC

Mol Biol 2009;10:71.

Alexandre HP, Caroline BM, Eric A, Florence BM, Frederic B, Cellier

Dominic C, et al. Reverse transcriptase quantitative polymerase chain

reaction: Description of a RIN-based algorithm for accurate data

normalization. BMC Mol Biol 2009;10:31.

Jebbink J, Bai X, Rogers BB, Dawson DB, Scheuermann RH, DomiatiSaad

R. Development of

real-time PCR

assays for the quantitative

detection

of

Epstein-Barr virus

and cytomegalovirus, comparison

of

TaqMan

probes,

and molecular beacons.

J

Mol

Diagn 2003;5(1):15-20.

Marum L, Miguel A, Ricardo CP, Miguel C. Reference gene selection

for quantitative real-time PCR normalization. Publ Lib Sci 2012;7:1371.

Guénin S, Mauriat M, Pelloux J, Van Wuytswinkel O, Bellini C,

Gutierrez L. Normalization of qRT-PCR data: The necessity of adopting

a systematic, experimental conditions-specific, validation of references.

J Exp Bot 2009;60(2):487-93.

Published

01-05-2016

How to Cite

SAS, N. ., H. H, S. . NNA, H. . AH, R. . SM, W. . SWZ, N. . ME, S. . S, N. . MM, and J. . S. “LOW LEVEL OF CYCLOOXYGENASE-2 TRANSCRIPT IN THE SPLEEN OF LYMPHOMA RATS SUPPLEMENTED WITH GARLIC POWDER”. Asian Journal of Pharmaceutical and Clinical Research, vol. 9, no. 3, May 2016, pp. 187-92, https://journals.innovareacademics.in/index.php/ajpcr/article/view/10965.

Issue

Section

Original Article(s)