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01-06-2009 | PRECLINICAL STUDIES

Nimbolide a limonoid from Azadirachta indica inhibits proliferation and induces apoptosis of human choriocarcinoma (BeWo) cells

Authors: G. Harish Kumar, K.V.P. Chandra Mohan, A. Jagannadha Rao, S. Nagini

Published in: Investigational New Drugs | Issue 3/2009

Summary

We investigated the cytotoxic effects of nimbolide, a limonoid present in leaves and flowers of the neem tree (Azadirachta indica) on human choriocarcinoma (BeWo) cells. Treatment with nimbolide resulted in dose- and time-dependent inhibition of growth of BeWo cells with IC₅₀ values of 2.01 and 1.19 μM for 7 and 24 h respectively, accompanied by downregulation of proliferating cell nuclear antigen. Examination of nuclear morphology revealed fragmentation and condensation indicating apoptosis. Increase in the generation of reactive oxygen species (ROS) that was reversed by addition of reduced glutathione suggested ROS involvement in the cytotoxicity of nimbolide. A decrease in Bcl-2/Bax ratio with increased expression of Apaf-1 and caspase-3, and cleavage of poly(ADP-ribose) polymerase provide compelling evidence that nimbolide-induced apoptosis is mediated by the mitochondrial pathway. The results of the present study suggest that nimbolide has immense potential in cancer prevention and therapy based on its antiproliferative and apoptosis inducing effects.

Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100:57–70 doi:10.1016/S0092-8674(00)81683-9 PubMedCrossRef

Lowe SW, Lin AW (2000) Apoptosis in cancer. Carcinogenesis 21:485–495 doi:10.1093/carcin/21.3.485 PubMedCrossRef

Reed JC (1999) Mechanisms of apoptosis avoidance in cancer. Curr Opin Oncol 11:68–75 doi:10.1097/00001622-199901000-00014 PubMedCrossRef

Vermeulen K, Van Bockstaele DR, Berneman ZN (2005) Apoptosis: mechanisms and relevance in cancer. Ann Hematol 84:627–639 doi:10.1007/s00277-005-1065-x PubMedCrossRef

Sun S-Y, Hail N, Lotan R Jr (2004) Apoptosis as a novel target for cancer chemoprevention. J Natl Cancer Inst 96:662–672PubMedCrossRef

Hsu S, Singh B, Schuster G (2004) Induction of apoptosis in oral cancer cells: agents and mechanisms for potential therapy and prevention. Oral Oncol 40:461–473 doi:10.1016/j.oraloncology.2003.09.012 PubMedCrossRef

Khan N, Afaq F, Mukhtar H (2008) Cancer chemoprevention through dietary antioxidants: progress and promise. Antioxid Redox Signal 10:475–510 doi:10.1089/ars.2007.1740 PubMedCrossRef

Davis CD (2007) Nutritional interactions: Credentialing of molecular targets for cancer prevention. Exp Biol Med 232:176–183

Deorukhkar A, Krishnan S, Sethi G et al (2007) Back to basics: how natural products can provide the basis for new therapeutics. Expert Opin Investig Drugs 16:1753–1773 doi:10.1517/13543784.16.11.1753 PubMedCrossRef

10.

Kang K, Lee HJ, Kim CY (2007) The chemopreventive effects of Saussurea salicifolia through induction of apoptosis and phase II detoxification enzyme. Biol Pharm Bull 30:2352–2359 doi:10.1248/bpb.30.2352 PubMedCrossRef

11.

Biswas K, Chattopadhyay I, Banerjee RK (2002) Biological activities and medicinal properties of neem (Azadirachta indica). Curr Sci 82:1336–1345

12.

Subapriya R, Nagini S (2005) Medicinal properties of neem leaves: A review. Curr Med Chem Anticancer Agents 5:149–156 doi:10.2174/1568011053174828 PubMedCrossRef

13.

Subapriya R, Kumaraguruparan R, Nagini S (2006) Expression of PCNA, cytokeratin, Bcl-2 and p53 during chemoprevention of hamster buccal pouch carcinogenesis by ethanolic neem (Azadirachta indica) leaf extract. Clin Biochem 39:1080–1087 doi:10.1016/j.clinbiochem.2006.06.013 PubMedCrossRef

14.

Subapriya R, Velmurugan B, Nagini S (2005) Modulation of xenobiotic-metabolizing enzymes by ethanolic neem leaf extract during hamster buccal pouch carcinogenesis. J Exp Clin Cancer Res 24:207–214

15.

Subapriya R, Nagini S (2003) Ethanolic neem leaf extract protects against N-methyl-N’-nitro-N-nitrosoguanidine-induced gastric carcinogenesis in Wistar rats. Asian Pac J Cancer Prev 4:215–223PubMed

16.

Manikandan P, Vidjaya Letchoumy P, Gopalakrishnan M et al (2008) Evaluation of Azadirachta indica leaf fractions for in vitro antioxidant potential and in vivo modulation of biomarkers of chemoprevention in the hamster buccal pouch carcinogenesis model. Food Chem Toxicol 46:2332–2343 doi:10.1016/j.fct.2008.03.013 PubMedCrossRef

17.

Roy MK, Kobori M, Takenaka M et al (2007) Antiproliferative effect on human cancer cell lines after treatment with nimbolide extracted from an edible part of the neem tree (Azadirachta indica). Phytother Res 21:245–250 doi:10.1002/ptr.2058 PubMedCrossRef

18.

Sastry BS, Suresh Babu K, Hari Babu T et al (2006) Synthesis and biological activity of amide derivatives of nimbolide. Bioorg Med Chem Lett 16:4391–4394 doi:10.1016/j.bmcl.2006.05.105 PubMedCrossRef

19.

Cohen E, Quistad GB, Casida JE (1996) Cytotoxicity of nimbolide, epoxyazadiradione and other limonoids from neem insecticide. Life Sci 58:1075–1081 doi:10.1016/0024-3205(96)00061-6 PubMedCrossRef

20.

Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63 doi:10.1016/0022-1759(83)90303-4 PubMedCrossRef

21.

Keum YS, Kim J, Lee KH et al (2002) Induction of apoptosis and caspase-3 activation by chemopreventive [6]-paradol and structurally related compounds in KB cells. Cancer Lett 177:41–47 doi:10.1016/S0304-3835(01)00781-9 PubMedCrossRef

22.

Wang IK, Lin-Shiau SY, Lin JK (1999) Induction of apoptosis by apigenin and related flavonoids through cytochrome c release and activation of caspase-9 and caspase-3 in leukaemia HL-60 cells. Eur J Cancer 35:1517–1525 doi:10.1016/S0959-8049(99)00168-9 PubMedCrossRef

23.

Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254 doi:10.1016/0003-2697(76)90527-3 PubMedCrossRef

24.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 doi:10.1038/227680a0 PubMedCrossRef

25.

Setzer WN, Setzer MC (2003) Plant-derived triterpenoids as potential antineoplastic agents. Mini Rev Med Chem 3:540–556 doi:10.2174/1389557033487854 PubMedCrossRef

26.

Mandal-Ghosh I, Chattopadhyay U, Baral R (2007) Neem leaf preparation enhances Th1 type immune response and anti-tumor immunity against breast tumor associated antigen. Cancer Immun 7:8PubMed

27.

Bose A, Haque E, Baral R (2007) Neem leaf preparation induces apoptosis of tumour cells by releasing cytotoxic cytokines from human peripheral mononuclear cells. Phytother Res 21:914–920 doi:10.1002/ptr.2185 PubMedCrossRef

28.

Suresh K, Suresh PK, Vijayababu MR et al (2006) Anticancer effects of ethanolic neem leaf extract on prostate cancer cell line (PC-3). J Ethnopharmacol 105:246–250 doi:10.1016/j.jep.2005.11.006 PubMedCrossRef

29.

Akudugu J, Gade G, Bohm L (2001) Cytotoxicity of azadirachtin A in human glioblastoma cell lines. Life Sci 68:1153–1160 doi:10.1016/S0024-3205(00)01017-1 PubMedCrossRef

30.

Nanduri S, Thunuguntla SSR, Nyavanandi VK et al (2003) Biological investigation and structure-activity relationship studies on azadirone from Azadirachta indica A. Juss. Bioorg Med Chem Lett 13:4111–4115 doi:10.1016/j.bmcl.2003.08.030 PubMedCrossRef

31.

Kigodi PG, Blasko G, Thebtaranonth Y et al (1989) Spectroscopic and biological investigation of nimbolide and 28-deoxonimbolide from Azadirachta indica. J Nat Prod 52:1246–1251 doi:10.1021/np50066a008 PubMedCrossRef

32.

Cohen E, Quisted GB, Jefferies PR (1996) Nimbolide is the principle cytotoxic component of neem seed insecticide preparations. Pestic Sci 48:135–140 doi: 10.1002/(SICI)1096-9063(199610)48:2>135::AID-PS451<3.0.CO;2-JCrossRef

33.

Moldovan GL, Pfander B, Jentsch S (2007) PCNA, the maestro of the replication fork. Cell 129:665–679 doi:10.1016/j.cell.2007.05.003 PubMedCrossRef

34.

Maga G, Hubscher U (2003) Proliferating cell nuclear antigen (PCNA): a dancer with many partners. J Cell Sci 116:3051–3060 doi:10.1242/jcs.00653 PubMedCrossRef

35.

Jeong S-Y, Seol D-W (2008) The role of mitochondria in apoptosis. BMB Rep 41:11–22PubMedCrossRef

36.

Youle RJ, Strasser A (2008) The Bcl-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9:47–59 doi:10.1038/nrm2308 PubMedCrossRef

37.

Mackey TJ, Borkowski A, Amin P et al (1998) Bcl-2/bax ratio as a predictive marker for therapeutic response to radiotherapy in patients with prostate cancer. Urology 52:1085–1090 doi:10.1016/S0090-4295(98)00360-4 PubMedCrossRef

38.

Roy MK, Kobori M, Takenaka M et al (2006) Inhibition of colon cancer (HT-29) cell proliferation by a triterpenoid isolated from Azadirachta indica is accompanied by cell cycle arrest and upregulation of p21. Planta Med 72:917–923 doi:10.1055/s-2006-946694 PubMedCrossRef

39.

Ralph SJ, Low P, Dong L et al (2006) Mitocans: mitochondrial targeted anti-cancer drugs as improved therapies and related patent documents. Recent Patents Anticancer Drug Discov 1:327–346CrossRef

Title: Nimbolide a limonoid from Azadirachta indica inhibits proliferation and induces apoptosis of human choriocarcinoma (BeWo) cells
Authors: G. Harish Kumar
K.V.P. Chandra Mohan
A. Jagannadha Rao
S. Nagini
Publication date: 01-06-2009
Publisher: Springer US
Published in: Investigational New Drugs / Issue 3/2009
Print ISSN: 0167-6997
Electronic ISSN: 1573-0646
DOI: https://doi.org/10.1007/s10637-008-9170-z

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