Study of the cytotoxicity of asiaticoside on rats and tumour cells

BMC Cancer

Fatma J Al-Saeedi¹ 

[1]Nuclear Medicine Department, Faculty of Medicine, Kuwait University, Al-Jabriya, Kuwait

关键词: Rats;    Apoptosis;    Proliferation;    Tumour;    DMBA;    Asiaticoside;   

Others  :  858937
DOI  :  10.1186/1471-2407-14-220

 received in 2013-02-27, accepted in 2014-03-13,  发布年份 2014

【 摘 要 】

Background

Cancer chemoprevention is considered one of the most promising areas in current cancer research, and asiaticoside, which is derived from the plant Centella asiatica, has a relative lack of systemic toxicity. The purpose of this study was to investigate whether asiaticoside is effective against 7,12-dimethylbenz(a)anthracene (DMBA)-induced carcinogenicity in vitro (MCF-7 and other cells) and in vivo (DMBA-induced rat cancer).

Methods

An MTT assay was performed involving the treatment of MCF-7 cells for 48 h with H₂O₂ alone and H₂O₂ + different asiaticoside concentrations. Flow cytometry was performed, and the level of caspase 3, tumour necrosis factor-alpha (TNF-α) and interleukin-1 (IL-1) were quantified. Adult female Sprague–Dawley (SD) rats were divided into five groups designated I (control), II (DMBA-induced cancer), III (pre- and post-treatment with asiaticoside (200 μg/animal) in DMBA-induced cancer), IV (post-treatment with asiaticoside in DMBA-induced cancer), and V (treated with asiaticoside alone, drug control). Twelve weeks post-DMBA, rats developed mammary tumours. Rats either were sacrificed or imaged with MIBI. Histological examination of tumour tissues was performed. Tumour MIBI uptake ratios were determined. The data are expressed as the means ± standard deviation. Appropriate t-test and ANOVA statistical methods were used to compare data.

Results

The IC50 of asiaticoside for MCF-7 cells was determined to be 40 μM. Asiaticoside has potential for hydrogen peroxide cytotoxicity, and the caspase-3 activity increased with increasing asiaticoside dose in MCF-7 cells treated for 48 h. The expression of the cytokines TNF-α and IL-1β was significantly decreased and correlated with MIBI uptake ratios in vitro and in vivo after asiaticoside administration.

Conclusion

This study demonstrates that asiaticoside is effective in vitro and in vivo in inducing apoptosis and enhancing anti-tumour activity.

【 授权许可】

   
2014 Al-Saeedi; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]World Health Organization, WHO: Cancer. Chapter 1 Burden. World Health Organization. http://www.who.int/nmh/publications/ncdreportchapter1.pdf webcite
• [2]Babu TD, Kuttan G, Padikkala J: Cytotoxic and antitumor properties of certain texa of umbelliferae with specific reference to Centella asiatica (L.) urban. J Ethnopharmacol 1995, 48:53-57.
• [3]Cheng CL, Guo JS, Luk J, Koo MW: The healing effects of Centella extract and asiaticoside on acetic acid induced gastric ulcers in rats. Life Sci 2004, 74:2237-2249.
• [4]Komarcević A: The modern approach to wound treatment. Med Pregl 2000, 53:363-368.
• [5]Suguna L, Sivakumar P, Chandrakasan G: Effects of centella asiatica extract on dermal wound healing in rats. Indian J Exp Biol 1996, 34:1208-1211.
• [6]Apparao MVR, Srinivasan K, Rao K: The effect of mandookparni (Centella asiatica) on the general mental ability (Medhya) of mentally retarded children. J Res Indian Med 1973, 8:9-16.
• [7]Oyedeji OA, Afolayan AJ: Chemical composition and antibacterial activity of the essential oil of Centella asiatica growing in South Africa. Pharm Biol 2005, 43:249-252.
• [8]Mook-Jung I, Shin JE, Yun SH, Huh K, Koh JY: Protective effects of asiaticoside derivatives against beta-amyloid neurotoxicity. J Neurosci Res 1999, 58:417-425.
• [9]Shukla A, Rasik AM, Dhawan BN: Asiaticoside-induced elevation of antioxidant levels in healing wounds. Phytother Res 1999, 13:50-54.
• [10]Salami S, Karami TF: Biochemical studies of apoptosis induced by tamoxifen in estrogen receptor positive and negative breast cancer cell lines. Clin Biochem 2003, 36:247-253.
• [11]Ma D, Zhang Y, Yang T, Xue Y, Wang P: Isoflavone intake inhibits the development of 7,12-dimethylbenz(a)anthracene(DMBA)-induced mammary tumors in normal and ovariectomized rats. J Clin Biochem Nutr 2014, 54:31-38. doi:10.3164/jcbn.13-33
• [12]Miyata M, Furukawa M, Takahashi K, Gonzalez FJ, Yamazoe Y: Mechanism of 7, 12-Dimethylbenz[a]anthracene-induced immunotoxicity: Role of metabolic activation at the target organ. Jpn J Pharmacol 2001, 86:302-309.
• [13]Kinuya S, Bai J, Shiba K, Yokoyama K, Mori H: 99mTc-sestamibi to monitor treatment with antisense oligodeoxynucleotide complementary to MRP mRNA in human breast cancer cells. Ann Nucl Med 2006, 20:29-34.
• [14]Aloj L, Zannetti A, Caracó C, Del Vecchio S, Salvatore M: Bcl-2 overexpression prevents 99mTc-MIBI uptake in breast cancer cell lines. Eur J Nucl Med Mol Imaging 2004, 31:521-527.
• [15]Rodrigues M, Chehne F, Kalinowska W, Berghammer P, Zielinski C: Uptake of 99mTc-MIBI and 99mTc-tetrofosmin into malignant versus nonmalignant breast cell lines. J Nucl Med 2000, 41:1495-1499.
• [16]de Jong M, Bernard BF, Breeman WA, Ensing G, Benjamins H: Comparison of uptake of 99mTc-MIBI, 99mTc-tetrofosmin and 99mTc-Q12 into human breast cancer cell lines. Eur J Nucl Med 1996, 23:1361-1366.
• [17]Al-Saeedi FJ, Bitar M, Pariyani S: Effect of asiaticoside on 99mTc-tetrofosmin and 99mTc-sestamibi uptake in MCF-7 cells. J Nucl Med Technol 2011, 39:279-283.
• [18]Piwnica-Worms D, Kronauge JF, Chiu ML: Uptake and retention of hexakis (2-methoxy isobutyl isonitrile) technetium (I) in cultured chick myocardial cells: mitochondrial and plasma membrane potential dependence. Circulation 1990, 82:1826. 1838
• [19]Piwnica-Worms D, Chiu ML, Budding J: Functional imaging of multi drug-resistant P-glycoprotein with an organo-technetium complex. Cancer Res 1993, 53:977-984.
• [20]Carlsson J, Nilsson K, Westermark B, Pontén J, Sundström C, Larsson E, Bergh J, Påhlman S, Busch C, Collins VP: Formation and growth of multicellular spheroids of human origin. Int J Cancer 1983, 31:523-533.
• [21]Huang YH, Zhang SH, Zhen RX, Xu XD, Zhen YS: Asiaticoside inducing apoptosis of tumor cells and enhancing anti-tumor activity of vincristine. Ai Zheng 2004, 23:1599-1604.
• [22]Gurfinkel DM, Chow S, Hurren R, Gronda M, Henderson C, Berube C, Hedley DW, Schimmer AD: Disruption of the endoplasmic reticulum and increases in cytoplasmic calcium are early events in cell death induced by the natural triterpenoid Asiatic acid. Apoptosis 2006, 11:1463-1471.
• [23]Gnanapragasam A, Yogeeta S, Subhashini R, Ebenezar KK, Sathish V, Devaki T: Adriamycin induced myocardial failure in rats: protective role of Centella asiatica. Mol Cell Biochem 2007, 294:55-63.
• [24]Wijeweera P, Arnason JT, Koszycki D, Merali Z: Evaluation of anxiolytic properties of Gotukola–(Centella asiatica) extracts and asiaticoside in rat behavioral models. Phytomedicine 2006, 13:668-676.
• [25]Harris RE, Alshafie GA, Abou-Issa H, Seibert K: Chemoprevention of breast cancer in rats by celecoxib, a cyclooxygenase 2 inhibitor. Cancer Res 2000, 60:2101-2103.
• [26]Ouhtit A, Ismail MF, Othman A, Fernando A, Abdraboh ME, El-Kott AF, Azab YA, Abdeen SH, Gaur RL, Gupta I, Shanmuganathan S, Al-Farsi YM, Al-Riyami H, Raj MH: Chemoprevention of rat mammary carcinogenesis by spirulina. Am J Pathol 2014, 184:296-303. doi: 10.1016/j.ajpath.2013.10.025
• [27]Hamdy SM, Latif AK, Drees EA, Soliman SM: Prevention of rat breast cancer by genistin and selenium. Toxicol Ind Health 2012, 28:746-757. doi:10.1177/0748233711422732
• [28]Kubatka P, Stollárová N, Škarda J, Žihlavníková K, Kajo K, Kapinová A, Adamicová K, Péč M, Dobrota D, Bojková B, Kassayová M, Orendáš P: Preventive effects of fluvastatin in rat mammary carcinogenesis. Eur J Cancer Prev 2013, 22:352-357. doi:10.1097/CEJ.0b013e32835b385d
• [29]Sharmila G, Athirai T, Kiruthiga B, Senthilkumar K, Elumalai P, Arunkumar R, Arunakaran J: Chemopreventive effect of quercetin in MNU and testosterone induced prostate cancer of Sprague–Dawley rats. Nutr Cancer 2014, 66:38-46. doi:10.1080/01635581.2014.847967
• [30]Huggins C, Briziarelli G, Sutton H: Rapid induction of mammary carcinoma in the rat and the influence of hormones on the tumors. J Exp Med 1959, 109:25-41.
• [31]Huggins C, Grand LC, Brillantes FP: Mammary cancer induced by a single feeding of polynuclear hydrocarbons, and its suppression. Nature 1961, 189:204-207.
• [32]Thompson HJ, McGinley JN, Rothhammer K, Singh M: Rapid induction of mammary intraductal proliferations, ductal carcinoma in situ and carcinomas by the injection of sexually immature female rats with 1-methyl-1-nitrosourea. Carcinogenesis 1995, 16:2407-2411.
• [33]Banerjee S, Bueso-Ramos C, Aggarwal BB: Suppression of 7,12-dimethylbenz(a)anthracene-induced mammary carcinogenesis in rats by resveratrol: role of nuclear factor-kappaB, cyclooxygenase 2, and matrix metalloprotease 9. Cancer Res 2002, 62:4945-4954.
• [34]Terada S, Uchide K, Suzuki N, Akasofu K, Nishida E: Induction of ductal carcinomas by intraductal administration of 7,12-dimethylbenz(a)anthracene in Wistar rats. Breast Cancer Res Treat 1995, 34:35-43.
• [35]Perumal BS, Sakharkar KR, Chow VT, Pandjassarame K, Sakharkar MK: Intron position conservation across eukaryotic lineages in tubulin genes. Front Biosci 2005, 10:2412-2419.
• [36]Padmavathi R, Senthilnathan P, Chodon D, Sakthisekaran D: Therapeutic effect of paclitaxel and propolis on lipid peroxidation and antioxidant system in 7,12 dimethyl benz (a) anthracene-induced breast cancer in female Sprague Dawley rats. Life Sci 2006, 8:2820-2825.
• [37]Shobi V, Goel HC: Protection against radiation-induced conditioned taste aversion by centella asiatica. Physiol Behav 2001, 73:19-23.
• [38]Sharma J, Sharma R: Radioprotection of Swiss albino mouse by centella asiatica extract. Phytother Res 2002, 16:785-786.
• [39]Horne T, Pappo I, Cohen-Pour M, Baumer M, Orda R: 99Tc(m)-tetrofosmin scintimammography for detecting breast cancer: a comparative study with 99Tc(m)-MIBI. Nucl Med Commun 2001, 22:807-811.
• [40]Söderlund V, Jonsson C, Bauer HC, Brosjö O, Jacobsson H: Comparison of technetium-99 m-MIBI and technetium-99 m-tetrofosmin uptake by musculoskeletal sarcomas. J Nucl Med 1997, 38:682-686.
• [41]Gao J, Huang F, Zhang J, Zhu G, Yang M: Cytotoxic cycloartane triterpene saponins from Actaea asiatica. J Nat Prod 2006, 69:1500-1502.
• [42]Beanlands RSB, Dawood F, Wen WH, McLaughlin PR, Butany J: Are the kinetics of technetium-99 m methoxyisobutyl isonitrile affected by cell metabolism and viability? Circulation 1990, 82:1802. 1814
• [43]Khalkhali I, Mena I, Jouanne E, Diggles L, Venegas R: Prone scintimammography in patients with suspicion of carcinoma of the breast. J Am Coll Surg 1994, 178:491-497.
• [44]Khalkhali I, Cutrone J, Mena I, Diggles L, Venegas R: Technetium-99 m sestamibi scintimammography of breast lesions: clinical and pathological follow-up. J Nucl Med 1995, 36:1784-1789.
• [45]Abdel Dayem MH, Scott AM, Macapinlac HA, El-Gazzar AH, Larson SM: Role of 201Tl chloride and 99mTc sestamibi in tumor imaging. In Nuclear Medicine Annual. Edited by Freeman LM. New York: Raven; 1994:181-234.
• [46]Palmedo H, Schomburg A, Grünwald F, Mallmann P, Krebs D: Technetium-99 m-MIBI scintimammography for suspicious breast lesions. J Nucl Med 1996, 37:626-630.
• [47]Pappo I, Horne T, Weissberg D, Wasserman I, Orda R: The usefulness of MIBI scanning to detect underlying carcinoma in women with acute mastitis. Breast J 2000, 6:126-129.
• [48]Grivennikov SI, Karin M: Inflammatory cytokines in cancer: tumour necrosis factor and interleukin 6 take the stage. Ann Rheum Dis 2011, 70:i104-i108. doi:10.1136/ard.2010.140145
• [49]Zheng J, Zhang L, Wu M, Li X, Zhang L, Wan J: Protective effects of asiaticoside on sepsis-induced acute kidney injury in mice. Zhongguo Zhong Yao Za Zhi 2010, 35:1482-1485.
• [50]Zhang Z, Qin DL, Wan JY, Zhou QX, Xiao SH: Effects of asiaticoside on the balance of inflammatory factors of mouse’s acute lung injury induced by LPS. Zhong Yao Cai 2008, 31:547-549.
• [51]Zhang LN, Zheng JJ, Zhang L, Gong X, Huang H: Protective effects of asiaticoside on septic lung injury in mice. Exp Toxicol Pathol 2011, 63:519-525.
• [52]Pan S, Li T, Li Y: Effects of asiaticoside on cell proliferation and Smad signal pathway of hypertrophic scar fibroblasts. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2004, 18:291-294.
• [53]Nowwarote N, Osathanon T, Jitjaturunt P, Manopattanasoontorn S, Pavasant P: Asiaticoside induces type I collagen synthesis and osteogenic differentiation in human periodontal ligament cells. Phytother Res 2013, 27:457-462. doi:10.1002/ptr.4742
• [54]Pizzorno JE, Murray MT: Textbook of Natural Medicine. London: Churchill Livingstone Press; 1999.
• [55]Boiteau P, Nigeon-Dureuil M, Ratsimamanga AR: Action of asiaticoside on the reticuloendothelial tissue. Acad Sci Compt Rend 1951, 232:760-762.