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5α-Oleandrin reduce Bcl-2 protein and increase Bax protein expression on Hela cervical cancer cell

Mae Sri Hartati Wahyuningsih, Sofia Mubarika, Ibnu G. Ganjar, Subagus Wahyuono, Tatsuo Takeya
Submission date: Monday, 29 May 2017
Published date: Monday, 07 August 2017
DOI: http://dx.doi.org/10.18051/UnivMed.2017.v36.102-109

Abstract


BACKGROUND

The leaves of Nerium indicum Mill. have been utilized traditionally to cure cancer. By using Bioassay guided extraction and isolation method, three compounds (NiO-1, NiO-2, NiO-3) were isolated from an active fraction of the CHCl3 extract of N. indicum leaves. NiO-2 identified as 5α- oleandrin is the best cytotoxic compound on HeLa cervical cancer cell in vitro among the other two. However mechanism of action of the compound hasn’t been evaluated yet. The aims of this study were to determine the mechanism of action at molecular level.

METHODS
The action mechanism of 5α-oleandrin on HeLa cervical cancer cells was analyzed by staining the cells with Hoechst 33342; the agarose gel electrophoresis was aimed to determine the DNA fragmentation and the western blotting was aimed to determine the Bcl-2 and Bax protein expression.

RESULTS
Incubation of HeLa cervical cancer cell with 5α-oleandrin at the concentration 3,47x10-4 mM (24 hours) followed by staining with Hoechst 33342, a broken up light blue color of nucleus was observed (compared with intensive color of untreated control). By gel electrophoresis (at the same concentration of the tested compound), a smear band at about 200 bp was observed. In addition, cells treated with 5α-oleandrin displayed a decreasing of the Bcl-2 protein expression and increasing of the Bax protein expression

CONCLUSION
5α-oleandrin induced HeLa cervical cancer cells DNA fragmentation observed by the presence of a smear band at about 200 bp indicative the apoptotic occurrence. 5α-oleandrin induces apoptosis by reducing the Bcl-2 protein expression but the Bax protein expression increases.


Keywords


Nerium indicum; 5α-oleandrin; cytotoxicity; DNA; anticancer

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References


Ferlay J, Soerjomataram I, Ervik M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 2014. doi:10.1002/ijc.29210 PMID:25220842.

Nuranna L, Donny NB, Purwoto G, et al. Prevalence, age distribution, and risk factors of visual inspection with acetic acid-positive from 2007 to 2011 in Jakarta. J Cancer Prev 2017;22: 103-7. doi: 10.15430/JCP.2017.22.2.103.

Aziz MF. Gynecological cancer in Indonesia. J Gynecol Oncol 2009;20:8-10.

Kumar S, Ahmad MK, Waseem M, et al. Drug targets for cancer treatment: an overview. Med Chem 2015;5:115-23. doi:10.4172/2161-0444. 1000252.

Ma Y, Zhu B, Yong L, et al. Regulation of intrinsic and extrinsic apoptotic pathways in osteosarcoma cells following Oleandrin treatment. Int J Mol Sci 2016;17:1950. doi: 10.3390/ijms17111950.

Callaway E. Famous HeLa human cell line gets its DNA sequenced. Nature 2013;500:132-3.

Vaughan L, Glanzel W, Korch C, et al. Widespread use of misidentified cell line KB (HeLa): incorrect attribution and its impact revealed through mining the scientific literature. Cancer Res 2017;77:2784-8. doi: 10.1158/0008-5472.

Newman RA, Yang P, Pawlus AD, et al. Cardiac glycosides as novel cancer therapeutic agents. Mol Interv 2008;8:36-49. doi: 10.1124/mi.8.1.8.

Botelhoa AFM, Mirandab AS, Jocab HC, et al. Hydroalcoholic extract from Nerium oleander L. (Apocynaceae) elicits arrhythmogenic activity. J Ethnopharmacol 2017;206:170–7. doi: 10.1016/j.jep.2017.05.031.

Yang CW, Chang HY, Hsu HY, et al. Identification of anti-viral activity of the cardenolides, Na+/K+-ATPase inhibitors, against porcine transmissible gastroenteritis virus. Toxicol Appl Pharmacol 2017. DOI: 10.1016/j.taap.2017.04.017.

Pan L, Zhang Y, Zhao W, et al. The Cardiac glycoside oleandrin induces apoptosis in human colon cancer cells via the mitochondrial pathway. Cancer Chemother Pharmacol 2017;80:91–100. Doi: 10.1007/s00280-017-3337-2.

Wahyuningsih MSH, Mubarika S, Bolhuis RLH, et al. Selectivity of compounds isolated from the leaves of Nerium indicum Mill. on various human cancer cell lines. Medical J Malaysia 2008;Suppl.A.63: 24-5.

Sonawane P, Choi YA, Pandya H, et al. Novel molecular multilevel targeted antitumor agents. Cancer Transl Med 2017;3:69–79.

Mishra T, Khullar M, Bhatia A. Anticancer potential of aqueous ethanol seed extract of Ziziphus mauritiana against cancer cell lines and Ehrlich ascites carcinoma. Evid Based Complement Altern Med 2011. Article ID 765029’11 pages. doi: 10.1155/2011/765029.

Hidayat A, Wiradisastra K, Hernowo BS, et al. Ekspresi Bcl-2 dan caspase-3 pascapaparan hipoksia hipobarik intermiten. MKB 2011;43: 166–70.

Flanagan L, Lindner AU, Chaumont CD, et al. BCL2 protein signalling determines acute responses to neoadjuvant chemoradiotherapy in rectal cancer. J Mol Med 2015;93:315–26.

Crowley LC, Marfell BJ, Waterhouse NJ. Analyzing Cell Death by Nuclear Staining with Hoechst 33342. Cold Spring Harb Protoc 2016;9. doi: 10.1101/pdb.prot087205.

Bao Z, Tian B, Wang X, et al. Oleandrin induces DNA damage responses in cancer cells by suppressing the expression of Rad51. Oncotarget 2016;7:59572-9.

Ma Y, Zhu B, Yong L, et al. Regulation of intrinsic and extrinsic apoptotic pathways in osteosarcoma cells following oleandrin treatment. Int J Mol Sci 2016;17. doi: 10.3390/ijms17111950.

Nolte E, Wach S, Silva IT, et al. A new semisynthetic cardenolide analog 3b-[2-(1-amantadine)-1-on-ethylamine]-digitoxigenin (AMANTADIG) affects G2/M cell cycle arrest and miRNA expression profiles and enhances proapoptotic survivin-2B expression in renal cell carcinoma cell lines. Oncotarget 2017;8:11676-91.


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