DOI: http://dx.doi.org/10.18203/2319-2003.ijbcp20162538

Studies on the antitumor potentials of betulinic acid against murine ascites Dalton’s lymphoma

Anamika Bhaumik, Surya Bali Prasad

Abstract


Background: Betulinic acid, a naturally occurring pentacyclic triterpene, exhibits a variety of biological activities including anticancer properties. Despite the wide importance of ethnobotanical studies on the anticancer therapeutic uses of betulinic acid, its exact role has not been fully elucidated. Therefore, the present studies were undertaken to evaluate the antitumor effect of betulinic acid in a murine malignant tumor model along with various biochemical changes in the tumor cells of the host.

Methods: Ascites Dalton’s lymphoma (DL) tumor-transplanted Swiss albino mice were treated with betulinic acid (i.p., 10 mg/kg body weight) and the pattern of host’s survival was analysed. The viability of DL cells was assessed using trypan blue exclusion test. The DL cells were also studied for the determination of apoptosis using fluorescence microscopy. Reduced glutathione and protein estimations were done in DL cells under different treatment conditions.

Results: Betulinic acid treatment caused a significant increase in life span (ILS ~150%) of the tumor-bearing hosts, which may indicate tumor regression/antitumor activity. Following betulinic acid treatment, decrease in DL cells viability and damaging changes in the cell membranes and a decrease in reduced glutathione content in DL cells were observed.

Conclusions: Present findings reveal the potent antitumor activity of betulinic acid against murine ascites Dalton’s lymphoma. The cytotoxicity of betulinic acid to build up antitumor effects may involve induction of apoptosis as well as a decrease in glutathione level in tumor cells.


Keywords


Betulinic acid, Dalton’s lymphoma, Antitumor activity, Apoptosis

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References


Boyle P, Levin B. World cancer report 2008, WHO, international agency for research on cancer (IARC). Eds; 2008:1-510.

Sikora K, Advani S, Koroltchouk V, Magrath I, Levy L, Pinedo H, et al. Essential drugs for cancer therapy: a world health organization consultation. Ann Oncol.1999;10:385-90.

Nygren P, Larsson R. Overview of the clinical efficacy of investigational anticancer drug. J Intern Med. 2003;253:46-75.

Newman DJ, Cragg GM, Snader KM. Natural products as sources of new drugs over the period 1981-2002. J Nat Prod. 2003;66(7):1022-37.

Ogura M, Cordell GA, Farnsworth NR. Constituents of Jacaranda caucana Pittier (Bignoniaceae), Lloydia. Potential anticancer agents. IV. 1977;40:157-68.

Sheth K, Jolad S, Wiedhopf R, Cole JR. Tumor-inhibitory agent from Hyptis emoryi (Labiatae). J Pharm Sci. 1972;61(11):1819.

Miles DH, Kokpol U, Zalkow LH, Steindel SJ, Nabors JB. Preliminary investigation of antitumor activity of Sarracenia flava. J Pharm Sci. 1974;63:613-5.

Trumbull ER, Bianchi E, Eckert DJ, Wiedhopf RM, Cole JR. Tumor inhibitory agents from Vauquelinia corymbosa (rosaceae). J Pharm Sci. 1976;65:1407-8.

Sandberg F, Duchevska K, Khristov V, Spasov S. Spondianthus preussii var. glaber Engler. Pharmacological screening and occurrence of triterpenes. Acta Pharm Suec. 1987;24:253-6.

Singh SS, Patro B, Tripathi V, Srivastava A, Pandey SC, Ghosh AC. J Med Arom Plant Sc. 2002; 24: 1031-7.

Tan YM, Yu R, Pezzuto JM. Betulinic acid-induced programmed cell death in human melanoma cells involves mitogen-activated protein kinase activation. Clin Cancer Res. 2003;9:2866-75.

Pisha E, Chai H, Lee IS, Chagwedera TE, Farnsworth NR, Cordell AC, et al. Discovery of betulinic acid as a selective inhibitor of human melanoma that functions by induction of apoptosis. Nat Med. 1995;1(10):1046-51.

Schuhly W, Heilmann J, Callis I, Sticher O. New triterpenoids with antibacterial activity from Zizyphus joazeiro. Planta Med. 1999;65(8):740-3.

Jagadeesh SG, Krupadanam GLD, Srimanna- rayana G, Murthy SS, Kaur A, Raja SS. Tobacco caterpillar antifeedant from the gotti stem wood triter-pene betulinic acid. J Agr Food Chem. 1998;46(7):2797-9.

Kashiwada Y, Wang HK, Nagao T, Kitanaka S, Yasuda I, Fujioka T, et al. Anti-AIDS agents. 30. Anti-HIV activity of oleanolic acid, pomolic acid, and structurally related triterpenoids. J Nat Prod. 1998;61(9):1090-5.

Chang CW, Wu TS, Hsieh YS, Kuo SC, Chao PDL. Terpenoids of Syzygium formosanum. J Nat Prod. 1999;62(2):327-8.

Higa M, Ogihara K, Yogi S. Bioactive naphtho- quinone derivatives from diospyros maritime blume. Chem Pharm Bull. 1998;46(8):1189-93.

Singh P, Sharma S. Triterpenoid constituents of the seed of Diospyros melanoxylon, Tecomella undulata, and Terminalia bellirica. J Indian Chem Soc. 1997;74:504-5.

Recio MC, Giner RM, Manez S, Gueho J, Julien HR, Hostettmann K, et al. Investigations on the steroidal anti-inflammatory activity of triterpenoids from Diospyros leucomelas. Planta Med. 1995;61(1):9-12.

Lin HC, Ding HY, Wu YC. Two novel com- pounds from Paeonia suffruticosa. J Nat Prod. 1998;61(3):343-6.

Zuco V, Supino R, Righetti SC, Cleris L, Marchesi E, Gambacorti-Passerini C, et al. Selective cytotoxicity of betulinic acid on tumor cell lines, but not on normal cells. Cancer Lett. 2002;175(1):17-25.

Pisha E, Chai H, Lee IS, Chagwedera TE, Fransworth NR, Cordell GA, et al. Discovery of betulinic acid as a selective inhibitor of human melanoma that functions by induction of apoptosis. Nat Med. 1995;1:1046-51.

Fulda S, Friesen C, Los M, Scaffidi C, Mier W, Benedict M, et al. Betulinic acid triggers CD95 (Apol/Fas)- and p53-independent apoptosis via activation of caspases in neuroectodermal tumors. Cancer Res.1997;57:4956-64.

Fulda S, Scaffidi C, Susin SA, Krammer PH, Kroemer G, Peter ME, et al. Activation of mitochondria and release of mitochondrial apoptogenic factors by betulinic acid. J Biol Chem. 1998;273:33942-8.

Fulda S, Jeremias I, Steiner HH, Pietsch T, Debatin KM. Betulinic acid: a new cytotoxic agent against malignant brain-tumor cells. Int J Cancer. 1999;82:435-41.

Schmidt ML, Kuzmanoff KL, Ling-Indeck L, Pezzuto JM. Betulinic acid induces apoptosis in human neuroblastoma cell lines. Eur J Cancer. 1997;33:2007-10.

Fulda S, Friesen C, Los M, Scaffidi C, Mier W, Benedict M, et al. Betulinic acid triggers CD95 (APO-1/Fas)- and p53-independent apoptosis via activation of caspases in neuroectodermal tumors. Cancer Res. 1997;57(21):4956-64.

Verma AK, Prasad SB. Changes in glutathione, oxidative stress and mitochondrial membrane potential in apoptosis involving the anticancer activity of cantharidin isolated from redheaded blister beetles, Epicauta hirticornis. Anticancer Agents Med Chem. 2013;13:1096-114.

Alitheen NB, Mashitoh AR, Yeap SK, Shuhaimi M, Abdul Manaf A, Nordin L. Cytotoxic effect of damnacanthal, nordamnacanthal, zerumbone and betulinic acid isolated from Malaysian plant sources. Int Food Res J. 2010;17:711-9.

Talwar GP. Handbook of Practical Immunology. National Book Trust, New Delhi; 1974:329.

Shylesh BS, Nair SA, Subramoniam A. Induction of cell specific apoptosis and protection from Dalton’s lymphoma challenge in mice by an active fraction from Emilia Sonchifolia. Indian J Pharmacol. 2005;37:232-7.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.

Sedlak J, Lindsay RH. Estimation of total protein-bound and non-protein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem. 1968;25:192-205.

Sriram MI, Mani Kanth SB, Kalishwaralal K, Gurunathan S. Antitumor activity of silver nanoparticles in Dalton’s lymphoma ascites tumor model. Int J Nanomed. 2010:753-62.

Higuchi Y. Chromosomal DNA fragmentation in apoptosis and necrosis induced by oxidative stress. Biochem Pharmacol. 2003;66(8):1527-35.

Pelicano H, Carney D, Huang P. ROS stress in cancer cells and therapeutic implications. Drug Resist Updat. 2004;7(2):97-110.

Gerster H. β-Carotene, vitamin E and vitamin C in different stages of experimental carcinogenesis. Eur J Clin Nutr. 1995;49(3):155-68.

Kroemer G, Reed JC. Mitochondrial control of cell death. Nat Med. 2000;6:513-9.

Dash SK, Chattopadhyay S, Ghosh T, Tripathy S, Das S, Das D, et al. Antileukemic efficacy of monomeric manganese-based metal complex on KG-1A and K562 cell lines. ISRN Oncol. 2013;1-10.