Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
권호 표지
DOI QR코드

DOI QR Code

Induction of Apoptosis by Methanolic Extract of Rubia Cordifolia Linn in HEp-2 Cell Line is Mediated by Reactive Oxygen Species

  • Shilpa, P.N. (Department of Biochemistry, University of Madras) ;
  • Sivaramakrishnan, V. (School of Chemical and Biotechnology, SASTRA University) ;
  • Devaraj, S. Niranjali (Department of Biochemistry, University of Madras)
  • Published : 2012.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Rubia cordifolia Linn, which belongs to the Rubiaceae family, is a well-known herb used in Ayurvedic medicine. In the present study, we investigated the influence of a methanolic extract (RC) on the induction of apoptosis in HEp-2 (human laryngeal carcinoma) cell line, as evidenced by cytotoxicity, morphological changes and modification in the levels of pro-oxidants. Inhibition of cell proliferation and lactate dehydrogenase (LDH) release increased in a time and dose-dependent manner. Further, reduced glutathione (GSH), glutathione transferase (GST) and protein levels decreased and lipid peroxidation increased significantly on RC treatment in a dose dependent manner when compared to controls. Based on the results we determined the optimal dose as 30mg/ ml and the apoptotic effect of RC extract (30 mg/ml) on HEp-2 cells was confirmed by fluorescent microscopy and transmission electron microscopy (TEM) based on morphological and ultrastructural changes. RC extract suppressed the proliferation of HEp-2 oral cancer cells inducing apoptotic cell death in vitro. These results point to potential of RC extract as an agent for the treatment of laryngeal squamous cell carcinoma.

Keywords

References

  1. Adwankar MK, Chitnis MP, Khandalekar DD, Bhadsavale CG (1980). Anticancer acvtivity of the extracts of Rubia cordifolia Linn. Ind J Exp Biol, 18, 102-9
  2. Chatelain K, Phippen S, McCabe J, et al (2011). Cranberry and grape seed extracts inhibit the proliferative phenotype of oral squamous cell carcinomas. Evid Based Complement Altern Med, ??, 1-12
  3. Cheon JH, Kim JS, Kim JM, et al (2006). Plant sterol guggulsterone inhibits nuclear factor-kB signaling in intestinal epithelial cells by blocking IkB kinase and ameliorates acute murine colitis. Inflamm Bowel Dis, 12, 1152-61. https://doi.org/10.1097/01.mib.0000235830.94057.c6
  4. David GFX, Hebert J, Wright CS (1973). The ultrastructure of the pineal ganglion in the ferret. J Anat, 115, 79-97.
  5. Day TA, Davis BK, Gillespie MB, et al (2003). Oral cancer treatment. Curr Treat Options Oncol, 4, 27-41. https://doi.org/10.1007/s11864-003-0029-4
  6. Dosseh C, Tessier AM, Delaveau P (1981). New quinones in Rubia cordifolia L. Roots, III. Planta Med, 43, 360-6. https://doi.org/10.1055/s-2007-971524
  7. Fotakis G, Timbrell JA (2006). In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett, 160, 171-7. https://doi.org/10.1016/j.toxlet.2005.07.001
  8. Freshney I (1994). Culture of Animal Cells: a manual of basic techniques. New York, John Wiley & Sons. Fourth Edition, 330-1
  9. Ghashm AA, Othman NH, Khattak MN, Ismail NM, Saini R (2010). Antiproliferative effect of Tualang honey on oral squamous cell carcinoma and osteosarcoma cell lines. BMC Complement Altern Med, 10, 49-57.
  10. Guimaraes A, Linden R (2004). Programmed cell deaths. Apoptosis and alternative deathstyles. Eur J Biochem, 271, 1638-50. https://doi.org/10.1111/j.1432-1033.2004.04084.x
  11. Habig WH, Pabst MJ, Jakoby WB (1974). Glutathione-Stransferase, the first enzymatic step in mercapturic acid formation. J Biol Chem, 249, 7130-139.
  12. 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, 461-73. https://doi.org/10.1016/j.oraloncology.2003.09.012
  13. Ichikawa H, Aggarwal BB (2006). Guggulsterone inhibits osteoclastogenesis induced by receptor activator of nuclear factor-$\beta$ ligand and by tumor cells by suppressing nuclear factor-$\beta$ activation. Clinc Cancer Res, 12, 662-8. https://doi.org/10.1158/1078-0432.CCR-05-1749
  14. Joharapurkar AA, Deode NM, Zambad SP, Umathe SN (2003). Immunomodulatory activity of alcoholic extract of Rubia cordifolia Linn. Indian Drugs, 40, 179-81.
  15. Kasture SB, Kasture VS, Chopde CT (2001) Anti-inflammatory activity of Rubia cordifolia roots. J Nat Remed, 1, 111-5.
  16. Kasture VS, Desmukh VK, Chopde CT (2000). Anticonvulsant and behavioral actions of triterpine isolated from Rubia cordifolia Linn. Ind J Exp Biol, 38, 675-80.
  17. Kim YA, Xiao D, Xiao H, et al (2007). Mitochondria-mediated apoptosis by diallyl trisulfide in human prostate cancer cells is associated with generation of reactive oxygen species and regulated by Bax/Bak. Mol Cancer Ther, 6, 1599-609. https://doi.org/10.1158/1535-7163.MCT-06-0754
  18. King M, Chatelain K, Farris D, et al (2007). Oral squamous cell carcinoma proliferative phenotype is modulated by proanthocyanidins: a potential prevention and treatment alternative for oral cancer. BMC Complement Altern Med, 7, 22-32. https://doi.org/10.1186/1472-6882-7-22
  19. Kummalue T, Charoenrat OO, Jiratchariyakul W, et al (2007). Antiproliferative effect of Erycibe elliptilimba on human breast cancer cell line. J Ethnopharm, 110, 439-43. https://doi.org/10.1016/j.jep.2006.10.025
  20. Mari M, Morales A, Colell A, Garci'a-Ruiz C, Fernandez- Checa JC (2009). Mitochondrial glutathione, a key survival antioxidant. Antioxid Redox signal, 11, 2685-700. https://doi.org/10.1089/ars.2009.2695
  21. Moron MS, De Pierre JW, Mannervik B (1979). Levels of glutathione, glutathione reductase and glutathione-Stransferase in rat lung and liver. Biochim Biophys Acta, 82, 67-70.
  22. Mosmann T (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65, 55-63. https://doi.org/10.1016/0022-1759(83)90303-4
  23. Nieland AA (1955). Lactic dehydrogenase of heart muscle. Methods in enzymology, 1, 449-54. https://doi.org/10.1016/0076-6879(55)01074-4
  24. Okhawa H, Ohishi N, Yagi K (1979). Assay of lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem, 95, 351-8. https://doi.org/10.1016/0003-2697(79)90738-3
  25. Patel PR, Nagari AA, Patel RC, Rathod DK, Patel VR (2011a). Invitro anticancer activity of Rubia cordifolia against Hela and Hep2 cell line. Int J Pharmacy Pharmcol Sci, 3, 70-1.
  26. Patel PR, Raval BP, Karanth HA, Patel VR (2011b). Potent antitumor activity of Rubia cordifolia. Int J Phytomed, 2, 44-6.
  27. Raju J, Patlolla JM, Swamy MV, Rao CV (2004). Diosgenin, a steroid saponin of Trigonella foenum graecum (Fenugreek), inhibits azoxymethane-induced aberrant crypt foci formation in F344 rats and induces apoptosis in HT-29 human colon cancer cells. Cancer Epidem Biomar, 13, 1392-8.
  28. Ramos AM, Aller P (2008). Quercetin decreases intracellular GSH content and potentiates the apoptotic action of the antileukemic drug arsenic trioxide in human leukemia cell lines. Biochem Pharmacol, 75, 1912-23. https://doi.org/10.1016/j.bcp.2008.02.007
  29. Samudio I, Konopleva M, Safe S, McQueen T, Andreeff M (2005). Guggulsterones induce apoptosis and differentiation in acute myeloid leukemia: identification of isomer-specific antileukemic activities of the pregnadienedione structure. Molecular Cancer Ther, 4, 1982-92. https://doi.org/10.1158/1535-7163.MCT-05-0247
  30. Sertoli A, Francalanci S, Giorgini S (1994) Prevention of allergic contact dermatitis with alternative products. Contact Dermatitis, 31, 322-3. https://doi.org/10.1111/j.1600-0536.1994.tb02029.x
  31. Shafi G, Munshi A, Hasan TN, et al (2009). Induction of apoptosis in HeLa cells by chloroform fraction of seed extracts of Nigella sativa. Cancer Cell Int, ??, 29-37.
  32. Shishodia S, Aggarwal BB (2004). Guggulsterone inhibits NFkappaB and IkappaBalpha kinase activation, suppresses expression of anti-apoptotic gene products, and enhances apoptosis. J Biol Chem, 279, 47148 -58. https://doi.org/10.1074/jbc.M408093200
  33. Sies H (1991). Oxidative stress: from basic research to clinical applications. Am J Med, 91, 31-8.
  34. Spanou C, Stagos D, Aligiannis N, Kouretas D (2010). Influence of potent antioxidant leguminosae family plant extracts on growth and antioxidant defense system of Hep2 cancer cell line. J Med Food, 13, 149-55. https://doi.org/10.1089/jmf.2009.0058
  35. Sridevi Sivakami PL, Nandhini C (2011). Extraction, isolation of medicinal plant pigments for natural food colours. Plant Arch, 11, 479-84.
  36. Stander BA, Marais S, Steynberg TJ, et al (2007). Influence of Sutherlandia frutescens extracts on cell numbers, morphology and gene expression in MCF-7 cells. J Ethnopharm, 112, 312-8. https://doi.org/10.1016/j.jep.2007.03.013
  37. Wilcock E, Hickman JA (1988). Characterisation of a $Na^{+}- K^{+}-CL-$ cotransporter in alkylating agents-sensitive L1210 murine leukemia cells. Biochim Biophys Acta, 946, 359-69. https://doi.org/10.1016/0005-2736(88)90411-7
  38. Wyllie AH (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature, 284, 555-6. https://doi.org/10.1038/284555a0
  39. Xiao D, Singh SV (2007). Phenethyl isothiocyanate inhibits angiogenesis in vitro and ex vivo. Cancer Res, 67, 2239-46. https://doi.org/10.1158/0008-5472.CAN-06-3645
  40. Xiao D, Singh SV (2008). z-Guggulsterone, a constituent of Indian Ayurvedic medicinal plant Commiphora mukul, inhibits angiogenesis in vitro and in vivo. Mol Cancer Ther, 7, 171-80. https://doi.org/10.1158/1535-7163.MCT-07-0491
  41. Xiao D, Vogel V, Singh SV (2006c). Benzyl isothiocyanateinduced apoptosis in human breast cancer cells is initiated by reactive oxygen species and regulated by Bax and Bak. Mol Cancer Ther, 5, 2931-45. https://doi.org/10.1158/1535-7163.MCT-06-0396

Cited by

  1. A Review of the Botany, Phytochemistry, Pharmacology and Toxicology of Rubiae Radix et Rhizoma vol.21, pp.12, 2016, https://doi.org/10.3390/molecules21121747
  2. Simultaneous Determination of Purpurin, Munjistin and Mollugin in Rat Plasma by Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry: Application to a Pharmacokinetic Study after Oral Administration of Rubia cordifolia L. Extract vol.21, pp.6, 2016, https://doi.org/10.3390/molecules21060717
  3. Biomimetic synthesis of silver nanoparticles from Streptomyces atrovirens and their potential anticancer activity against human breast cancer cells vol.11, pp.8, 2017, https://doi.org/10.1049/iet-nbt.2016.0222
  4. FFJ-3 inhibits PKM2 protein expression via the PI3K/Akt signaling pathway and activates the mitochondrial apoptosis signaling pathway in human cancer cells vol.13, pp.4, 2017, https://doi.org/10.3892/ol.2017.5761
  5. Hydrogen peroxide-induced mitophagy contributes to laryngeal cancer cells survival via the upregulation of FUNDC1 pp.1699-3055, 2018, https://doi.org/10.1007/s12094-018-1958-5