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BMC Complementary Medicine and Therapies
Published in: BMC Complementary Medicine and Therapies 1/2016

Open Access 01-12-2016 | Research article

Comparison of cytotoxicity between extracts of Clinacanthus nutans (Burm. f.) Lindau leaves from different locations and the induction of apoptosis by the crude methanol leaf extract in D24 human melanoma cells

Authors: Siat Yee Fong, Terrence Piva, Chaitali Dekiwadia, Sylvia Urban, Tien Huynh

Published in: BMC Complementary Medicine and Therapies | Issue 1/2016

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Abstract

Background

Clinacanthus nutans (Burm. f.) Lindau leaves are widely used by cancer patients and the leaf extracts possess cytotoxic and antiproliferative effects on several human cancer cell lines. However, the effect of C. nutans leaf extract on human melanoma, which is the least common but most fatal form of skin cancer and one of the most common cancers diagnosed in both sexes worldwide, is unknown. There is also limited information on whether the bioactivity of extracts differs between C. nutans leaves grown in different geographical locations with varying environmental conditions.

Methods

The present study, for the first time, compared and demonstrated the cytotoxicity of the crude methanol extracts of C. nutans leaves from 11 different locations in Malaysia, Thailand and Vietnam, with diverse environmental conditions against D24 melanoma cells through WST-8 assay. The percentage of apoptotic cells following treatment with the most active extract was determined in a dose- and time-dependent manner by a cytofluorometric double staining technique. Biochemical and morphological changes in the treated and untreated cells were examined by fluorescence and transmission electron microscopy techniques, respectively, to further affirm the induction of apoptosis.

Results

The leaves of plants grown at higher elevations and lower air temperatures were more cytotoxic to the D24 melanoma cells than those grown at lower elevations and higher air temperatures, with the leaf extract from Chiang Dao, Chiang Mai, Thailand exhibited the highest cytotoxicity (24 h EC50: 0.95 mg/mL and 72 h EC50: 0.77 mg/mL). This most active crude extract induced apoptotic cell death in the D24 cells in a dose- and time-dependent manner. Typical biochemical and morphological characteristics of apoptosis were also observed in the treated D24 cells.

Conclusions

The results, showing the cytotoxicity of C. nutans and the induction of apoptosis in D24 cells, are significant and useful to facilitate the development of C. nutans as a potential novel chemotherapeutic agent for the management of skin melanoma.
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Literature
1.
Forman D, Ferlay J. The global and regional burden of cancer. In: Stewart BW, Wild CP, editors. World cancer report 2014. 1st ed. Lyon, France: International Agency for Research on Cancer; 2014. p. 16–53.
2.
3.
Cifola I, Pietrelli A, Consolandi C, Severgnini M, Mangano E, Russo V, Bellis GD, Battaglia C. Comprehensive genomic characterization of cutaneous malignant melanoma cell lines derived from metastatic lesions by whole-exome sequencing and SNP array profiling. PLoS ONE. 2013;8:e63597.CrossRefPubMedPubMedCentral
4.
Ferlay J, Soerjomataram I, Ervik M, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F. GLOBOCAN 2012 v1.0, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 11 International Agency for Research on Cancer, Lyon. 2013. http://​globocan.​iarc.​fr/​. Accessed 27 June 2015.
5.
Batus M, Waheed S, Ruby C, Petersen L, Bines S, Kaufman H. Optimal management of metastatic melanoma: current strategies and future directions. Am J Clin Dermatol. 2013;14:179–94.CrossRefPubMedPubMedCentral
6.
7.
Iyer AK, Singh A, Ganta S, Amiji MM. Role of integrated cancer nanomedicine in overcoming drug resistance. Adv Drug Deliv Rev. 2013;65:1784–802.CrossRefPubMed
8.
Kunjachan S, Rychlik B, Storm G, Kiessling F, Lammers T. Multidrug resistance: physiological principles and nanomedical solutions. Adv Drug Deliv Rev. 2013;65:1852–65.CrossRefPubMedPubMedCentral
9.
Markman JL, Rekechenetskiy A, Holler E, Ljubimova JY. Nanomedicine therapeutic approaches to overcome cancer drug resistance. Adv Drug Deliv Rev. 2013;65:1866–79.CrossRefPubMed
10.
Pratheeshkumar P, Sreekala C, Zhang Z, Budhraja A, Ding S, Son YO, Wang X, Hitron A, Hyun-Jung K, Wang L, Lee JC, Shi X. Cancer prevention with promising natural products: mechanisms of action and molecular targets. Anticancer Agents Med Chem. 2012;12:1159–84.CrossRefPubMedPubMedCentral
11.
12.
Chinembiri TN, du Plessis LH, Gerber M, Hamman JH, du Plessis J. Review of natural compounds for potential skin cancer treatment. Molecules. 2014;19:11679–721.CrossRefPubMed
13.
Wanikiat P, Panthong A, Sujayanon P, Yoosook C, Rossi AG, Reutrakul V. The anti-inflammatory effects and the inhibition of neutrophil responsiveness by Barleria lupulina and Clinacanthus nutans extracts. J Ethnopharmacol. 2008;116:234–44.CrossRefPubMed
14.
Pannangpetch P, Laupattarakasem P, Kukongviriyapan V, Kukongviriyapan U, Kongyingyoes B, Aromdee C. Antioxidant activity and protective effect against oxidative hemolysis of Clinacanthus nutans (Burm. F.) Lindau. Songklanakarin J Sci Technol. 2007;29:1–9.
15.
Yong YK, Tan JJ, Teh SS, Mah SH, Ee GCL, Chiong HS, Ahmad Z. Clinacanthus nutans extracts are antioxidant with antiproliferative affect on cultured human cancer cell lines. Evid Based Complement Alternat Med. 2013;2013:1–8.CrossRef
16.
Arullappan S, Rajamanickam P, Thevar N, Kodimani CC. In vitro screening of cytotoxic, antimicrobial and antioxidant activities of Clinacanthus nutans (Acanthaceae) leaf extracts. Trop J Pharm Res. 2014;13:1455–61.CrossRef
17.
Tiew WP, P'ng XW, Chin JH, Akowuah GA. Effect of methanol extract of Clinacanthus nutans on serum biochemical parameters in rats. J Appl Pharm. 2014;6:77–86.
18.
Wong FC, Yong AL, Ting EPS, Khoo SC, Ong HC, Chai TT. Antioxidant, metal chelating, anti-glucosidase activities and phytochemical analysis of selected tropical medicinal plants. Iranian J Pharm Res. 2014;13:1409–15.
19.
Thongchai S, Ekalaksananan T, Pientong C, Aromdee C, Seubsasana S, Sukpol C, Kongyingyoes B. Anti-herpes simplex virus type 1 activity of crude ethyl acetate extract derived from leaves of Clinacanthus nutans Lindau. J Sci Technol Mahasarakham U. 2008;27:318–26.
20.
Sakdarat S, Shuyprom A, Pientong C, Ekalaksananan T, Thongchai S. Bioactive constituents from the leaves of Clinacanthus nutans Lindau. Bioorg Med Chem. 2009;17:1857–60.CrossRefPubMed
21.
Kunsorn P, Ruangrungsi N, Lipipun V, Khanboon A, Rungsihirunrat K. The identities and anti-herpes simplex virus activity of Clinacanthus nutans and Clinacanthus siamensis. Asian Pac J Trop Biomed. 2013;3:284–90.CrossRefPubMedPubMedCentral
22.
Vachirayonstien T, Promkhatkaew D, Bunjob M, Chueyprom A, Chavalittumrong P, Sawanpanyalert P. Molecular evaluation of extracellular activity of medicinal herb Clinacanthus nutans against herpes simplex virus type-2. Nat Prod Res. 2010;24:236–45.CrossRefPubMed
23.
Thawaranantha D, Balachandra K, Jongtrakulsiri S, Chavalittumrong P, Bhumiswasdi J, Janyavasu C. In vitro antiviral activity of Clinacanthus nutans on varicellazoster virus. Siriraj Hosp Gaz. 1992;44:285–91.
24.
Sookmai W, Ekalaksananan T, Pientong C, Sakdarat S, Kongyingyoes B. The anti-papillomavirus infectivity of Clinacanthus nutans compounds. Srinagarind Med J. 2011;26:240–3.
25.
Tu SF, Liu RH, Cheng YB, Hsu YM, Du YC, El-Shazly M, Wu YC, Chang FR. Chemical constituents and bioactivities of Clinacanthus nutans aerial parts. Molecules. 2014;19:20382–90.CrossRefPubMed
26.
Hijmans RJ, Guarino L, Mathur P. DIVA-GIS. 7.5 ed. University of California Davis, California; 2012.
27.
Gatan. Gatan Microscopy Suite. DigitalMicrograph. 2.3 ed. Pleasanton, California: Gatan Inc.; 2013.
28.
Minitab. Minitab statistical software. 17 ed. State College, Pennsylvania: Minitab Inc; 2013.
29.
Akula R, Ravishankar GA. Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behav. 2011;6:1720–31.CrossRef
30.
Radušienė J, Karpavičienė B, Stanius Ž. Effect of external and internal factors on secondary metabolites accumulation in St. John’s worth. Bot Lith. 2012;18:101–8.
31.
Ayob Z, Mohd Bohari SP, Abd Samad A, Jamil S. Cytotoxic activities against breast cancer cells of local Justicia gendarussa crude extracts. Evid Based Complement Alternat Med. 2014;2014:1–12.CrossRef
32.
Basar N, Oridupa OA, Ritchie KJ, Nahar L, Osman NM, Stafford A, Kushiev H, Kan A, Sarker SD. Comparative cytotoxicity of Glycyrrhiza glabra roots from different geographical origins against immortal human keratinocyte (HaCaT), lung adenocarcinoma (A549) and liver carcinoma (HepG2) cells. Phytother Res. 2015;29:944–8.CrossRefPubMed
33.
McLachlan A, Kekre N, McNulty J, Pandey S. Pancratistatin: a natural anti-cancer compound that targets mitochondria specifically in cancer cells to induce apoptosis. Apoptosis. 2005;10:619–30.CrossRefPubMed
34.
Rodrigues MF, Obre E, de Melo FH, Santos Jr GC, Galina A, Jasiulionis MG, Rossignol R, Rumjanek FD, Amoedo ND. Enhanced OXPHOS, glutaminolysis and beta-oxidation constitute the metastatic phenotype of melanoma cells. Biochem J. 2016;473:703–15.CrossRefPubMed
35.
Piva TJ, McEvoy-Bowe E. Oxidation of glutamine in HeLa cells: role and control of truncated TCA cycles in tumour mitochondria. J Cell Biochem. 1998;68:213–25.CrossRefPubMed
36.
Liu T, Hannafon B, Gill L, Kelly W, Benbrook D. Flex-Hets differentially induce apoptosis in cancer over normal cells by directly targeting mitochondria. Mol Cancer Ther. 2007;6:1814–22.CrossRefPubMedPubMedCentral
37.
Li YX, Zhu JX, Yang HX, Yuan K. Studies on antitumor activities of six glycosylflavones from Mimosa pudica. 2011 International Symposium on IT in Medicine and Education (ITME 2011); 2011 9–11. Guangzhou, China: Institute of Electrical and Electronics Engineers (IEEE); 2011.
38.
Zhang Y, Tang H, Li A, Xu L, Chen J, Huang S, He L. Determination of six C-glycoside flavones and antitumor activity of water-soluble total flavonoids from Isodon lophanthoides var. gerardianus. Zhongguo Zhong Yao Za Zhi. 2015;40:1543–7.PubMed
39.
Brauchle E, Thude S, Brucker SY, Schenke-Layland K. Cell death stages in single apoptotic and necrotic cells monitored by Raman microspectroscopy. Sci Rep. 2014;4:1–9.CrossRef
40.
41.
Chen Y, Sun R, Han W, Zhang Y, Song Q, Di C, Ma D. Nuclear translocation of PDCD5 (TFAR19): an early signal for apoptosis? FEBS Lett. 2001;509:191–6.CrossRefPubMed
42.
Ziegler U, Groscurth P. Morphological features of cell death. News Physiol Sci. 2004;19:124–8.PubMed
43.
Svejda B, Aguiriano-Moser V, Sturm S, Hoger H, Ingolic E, Siegl V, Stuppner H, Pfragner R. Anticancer activity of novel plant extracts from Trailliaedoxa gracilis (W. W. Smith & Forrest) in human carcinoid KRJ-I cells. Anticancer Res. 2010;30:55–64.PubMed
44.
Zeiss CJ. The apoptosis-necrosis continuum: insights from genetically altered mice. Vet Pathol. 2003;40:481–95.CrossRefPubMed
45.
Cáceres-Cortés JR, Cantú-Garza FA, Mendoza-Mata MT, Chavez-González MA, Ramos-Mandujano G, Zambrano-Ramírez IR. Cytotoxic activity of Justicia spicigera is inhibited by bcl-2 proto-oncogene and induces apoptosis in a cell cycle dependent fashion. Phytother Res. 2001;15:691–7.CrossRefPubMed
46.
Alonso-Castro AJ, Ortiz-Sánchez E, Domínguez F, Arana-Argáez V, Juárez-Vázquez MC, Chávez M, Carranza-Álvarez C, Gaspar-Ramírez O, Espinosa-Reyes G, López-Toledo G, Ortiz-Andrade R, García-Carrancá A. Antitumor and immunomodulatory effects of Justicia spicigera Schltdl (Acanthaceae). J Ethnopharmacol. 2012;141:888–94.CrossRefPubMed
47.
Dey S, Roy S, Deb N, Sen KK, Besra SE. Anti-carcinogenic activity of Ruellia tuberosa L. (Acanthaceae) leaf extract on hepatoma cell line & increased superoxide dismutase activity on macrophage cell lysate. Int J Pharm Pharm Sci. 2013;5:854–61.
48.
Saengkhae C, Meewassanasuk K. Antiproliferation and apoptosis of the crude extract of Andrographis paniculata Nees, on human oropharyngeal cancer cells (KB) in vitro. Thai J Physiol Sci. 2009;21:40–7.
49.
Konur A, Schulz U, Eissner G, Andreesen R, Holler E. Interferon (IFN)-gamma is a main mediator of keratinocyte (HaCaT) apoptosis and contributes to autocrine IFN-gamma and tumour necrosis factor-alpha production. Br J Dermatol. 2005;152:1134–42.CrossRefPubMed
50.
Thongrakard V, Tencomnao T. Modulatory effects of Thai medicinal plant extract on proinflammatory cytokines-induced apoptosis in human keratinocyte HaCaT cells. Afr J Biotechnol. 2010;9:4999–5003.
51.
Hirsch T, Marchetti P, Susin SA, Dallaporta B, Zamzami N, Marzo I, Geuskens M, Kroemer G. The apoptosis-necrosis paradox. Apoptogenic proteases activated after mitochondrial permeability transition determine the mode of cell death. Oncogene. 1997;15:1573–81.CrossRefPubMed
52.
Ahmad NH, Rahim RA, Mat I. Catharanthus roseus aqueous extract is cytotoxic to jurkat leukaemic T-cells but induces the proliferation of normal peripheral blood mononuclear cells. Trop Life Sci Res. 2010;21:101–13.PubMedPubMedCentral
53.
Cock IE. Problems of reproducibility and efficacy of bioassays using crude extratcs, with reference to Aloe vera. Phcog Commn. 2011;1:52–62.CrossRef
Metadata
Title
Comparison of cytotoxicity between extracts of Clinacanthus nutans (Burm. f.) Lindau leaves from different locations and the induction of apoptosis by the crude methanol leaf extract in D24 human melanoma cells
Authors
Siat Yee Fong
Terrence Piva
Chaitali Dekiwadia
Sylvia Urban
Tien Huynh
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2016
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-016-1348-x

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