DOI QR코드

DOI QR Code

Purification of Lovastatin from Aspergillus terreus (KM017963) and Evaluation of its Anticancer and Antioxidant Properties

  • Bhargavi, SD (Department of Microbiology and Biotechnology, Bangalore University) ;
  • Praveen, VK (Department of Microbiology and Biotechnology, Bangalore University) ;
  • Marium, Salah (Department of Microbiology and Biotechnology, Bangalore University) ;
  • Sreepriya, M (Department of Microbiology and Biotechnology, Bangalore University) ;
  • Savitha, J (Department of Microbiology and Biotechnology, Bangalore University)
  • Published : 2016.08.01

Abstract

Cervical cancer is the second most common malignancy in women worldwide and thus one of the leading causes of mortality in women. Lovastatin, a non polar, anticholesterol drug has previously been reported to exert antitumour activity in vitro. In the present study, lovastatin from Aspergillus terreus (KM017963) was purified by adsoprtion chromatography and evaluated for its anticancer and anti-oxidant properties with a human cervical cancer cell line (HeLa). Growth inhibitory and proapoptotic effects of purified lovastatin on HeLa cells were investigated by determining its influence on cell numbers, mitochondrial membrane potential (MMP), DNA fragmentation and antioxidant properties in terms of hydroxy radical scavenging effects as well as levels of total reduced glutathione. Cell cycle analysis by flow cytometry (propidium iodide staining) confirmed induction of apoptotic cell death and revealed cell cycle arrest in the G0/G1 phase. Results of the study give leads for the anticancer effects of lovastatin and its potential usefulness in the chemotherapy of cervical cancer.

Keywords

Purification;lovastatin;Aspergillus terreus;proapoptotic effects;antioxidant activity

Acknowledgement

Supported by : Department of Science & Technology (DST)

References

  1. Agarwal B, Bhendwal S, Halmos B (1999). Lovastatin augments apoptosis induced by hemotherapeutic agents in colon cancer cells. Clin Cancer Res, 5, 2223-9.
  2. Ahmad A, Panda BP, Khan S, Ali M, Javed S (2009). Down streaming and purification of lovastatin from Monascus purpureus culture. Thai J Pharm Sci, 33, 39-6.
  3. Ajith, TA, et al (2006). Proapoptotic and antitumor activities of the HMG-CoA reductase inhibitor, lovastatin, against Dalton’s lymphoma ascites tumor in mice. Clin Chim Acta, 366, 322-8. https://doi.org/10.1016/j.cca.2005.11.012
  4. Bishayee K, et al (2013). Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M, in cervical carcinoma: signal cascade and drug-DNA interaction. Cell Proliferat, 46, 153-63. https://doi.org/10.1111/cpr.12017
  5. Linda BB, et al (2010). Lovastatin prevents carcinogenesis in a rat model for liver cancer. Effects of ubiquinone supplementation. Anticancer Res, 30, 1105-2.
  6. Thorskov BT (2013). Anticancer and antifungal compounds from Aspergillus, Penicillium and other filamentous fungi. Molecules, 18, 11338-6. https://doi.org/10.3390/molecules180911338
  7. Lucie B, Desgagne A, LeLorier J (2000). 3-Hydroxy-3- methylglutaryl coenzyme A reductase inhibitors and the risk of cancer: a nested case-control study. Archives Int Med, 160, 2363-8. https://doi.org/10.1001/archinte.160.15.2363
  8. Choi JW, Jung SE (1999). Lovastatin-induced proliferation inhibition and Leukemia apoptosis in C6 glial cells. J Pharmacol Exp Ther, 289, 572-9.
  9. Elena CM, et al (2009). Statins inhibit the proliferation and induce cell death of human papilloma virus positive and negative cervical cancer cells. IJBS, 5, 411-0.
  10. Francois D, Lang R (1986). Rapid colorimetric assay for cell growth and survival: modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunological Methods, 89, 271-7. https://doi.org/10.1016/0022-1759(86)90368-6
  11. Dimitroulakas J, Thai S, Wasfy GH (2000). Lovastatin induces a pronounced differentiation response in acute myeloid leukemias. Leuk Lymphoma, 40, 167-8. https://doi.org/10.3109/10428190009054894
  12. Ellman GL (1959). Tissue sulfhydryl groups. Arch Biochem Biophys, 82, 70-7. https://doi.org/10.1016/0003-9861(59)90090-6
  13. Ferdinando F, et al (2003). A comparative study of the in vitro antioxidant activity of statins. Int J Cardiol, 90, 317-1. https://doi.org/10.1016/S0167-5273(02)00577-6
  14. Toshihiko F, et al (2010). Tumor specific low pH environments enhance the cytotoxicity of lovastatin and cantharidin. Cancer Letters. 297, 182-9. https://doi.org/10.1016/j.canlet.2010.05.010
  15. Gey GO, Coffman WD, Kubicek MT (1952). Tissue culture studies of the proliferative capacity of cervical carcinoma and normal epithelium. Cancer Res, 12, 264-5.
  16. Gotto AM, Farmer JA (2001). Pleiotropic effects of statins: do they matter? Curr Opin Lipidol, 12, 391-4. https://doi.org/10.1097/00041433-200108000-00004
  17. Julie GB, Rao S, and Keyomarsi K (1997). Lovastatin induction of cyclin-dependent kinase inhibitors in human breast cells occurs in a cell cycle-independent fashion. Cancer Res, 57, 604-9.
  18. Gulyamova TG, Ruzieva DM, Nasmetova SM, et al (2013). Lovastatin production by Aspergillus terreus in solid state and submerged fermentations. Int J Eng Sci Tech, 5, 19-4.
  19. Barry H, Grootveld M (1987). The measurement of free radical reactions in humans: some thoughts for future experimentation. FEBS letters, 213, 9-4. https://doi.org/10.1016/0014-5793(87)81455-2
  20. Joany J, O'Connor PM (2001). Methods for synchronizing cells at specific stages of the cell cycle. Current Protocols Cell Biol, 8-3.
  21. Jafari M, Rezaei M, Kalantari H, Hashemitabar M (2013). Determination of cell death induced by lovastatin on human colon cell line HT29 using the comet assay. Jundishapur J Natural Pharmaceutical Products, 8, 187-1. https://doi.org/10.17795/jjnpp-10951
  22. Jakobisiak M, Bruno S, Skierski JS, Darzynkiewicz Z (1991). Cell cycle-specific effects of lovastatin. Proceed National Academy Sci, 88, 3628-2. https://doi.org/10.1073/pnas.88.9.3628
  23. Johnson LV, Walsh ML., Bockus BJ, Chen LB (1981). Monitoring of relative mitochondrial membrane potential in living cells by fluorescence microscopy. J Cell Biol, 88, 526-5. https://doi.org/10.1083/jcb.88.3.526
  24. Landry JJM, Pyl PT, Rausch T, et al (2013). The genomic and transcriptomic landscape of a HeLa cell line. G3: Genes Genomes Genetics, 3, 1213-4.
  25. Laurie M, et al (2012). Lovastatin induces multiple stress pathways including LKB1/AMPK activation that regulate its cytotoxic effects in squamous cell carcinoma cells. PloS one, 7, 46055. https://doi.org/10.1371/journal.pone.0046055
  26. Macreadie IG, Johnson G, Schlosser T, Macreadie PI, et al (2006). Growth inhibition of Candida species and Aspergillus fumigatus by statins. FEMS Microbiol Letters, 262, 9-13. https://doi.org/10.1111/j.1574-6968.2006.00370.x
  27. Maltese WA, Defendini R, Green RA, et al (1985). Suppression of murine neuroblatoma growth invivo by mevinolin, a competitive inhibitor of 3-hydroxy-3-methylglutarylcoenzyme A reductase. J Clin Invest, 5, 1748-4.
  28. Matsuyama S, Reed JC (2000). Mitochondria-dependent apoptosis and cellular pH regulation. Cell Death And Differentiat, 7, 1155-5. https://doi.org/10.1038/sj.cdd.4400779
  29. MohanKumari HP, Dhale MA, Naidu AK, Vijayalakshmi G (2011). Antioxidant effect of red mould rice in hypercholesterolemic Wistar male rats. Cell Biochem Funct, 29, 597-2. https://doi.org/10.1002/cbf.1793
  30. Muller C, Bockhorn AG, Klusmeier S, et al (1998). Lovastatin inhibits proliferation of pancreatic cancer cell lines with mutant as well as with wild type K-ras oncogene but has different effects o protein phosphorylation and induction of apoptosis. Int J Oncol, 12, 717-3.
  31. Jaivel N, Marimuthu P (2010). Optimization of lovastatin production in solid state fermentation by Aspergillus terreus. Intl J Engg Sci Technol, 2, 2730-3
  32. Poon, RY., Toyoshima H, and Hunter T (1995). Redistribution of the CDK inhibitor p27 between different cyclin. CDK complexes in the mouse fibroblast cell cycle and in cells arrested with lovastatin or ultraviolet irradiation. Molecular Biol Cell, 6, 1197-3. https://doi.org/10.1091/mbc.6.9.1197
  33. Praveen VK, Bhargavi SD, Savitha J (2014). Endophytic fungi: a poor candidate for the production of lovastatin. B Microbiol Res J, 4, 1511-0. https://doi.org/10.9734/BMRJ/2014/12487
  34. Raghunath R, Radhakrishna A, Angayarkanni J, Palaniswamy M (2012). Production and cytotoxicity studies of lovastatin from Aspergillus niger PN2 an endophytic fungi isolated from Taxus baccata. Ijabpt, 3, 342-1
  35. Reddy SR, Latha DP, Latha KPJ (2011). Production of lovastatin by solid state fermentation by Penicillium funiculosum NCIM 1174. Drug Invention Today, 3, 75-7.
  36. Samiee SM, Moazami N, Haghighi S, et al (2003). Screening of lovastatin production by filamentous fungi. Iran Biomed J, 7, 29-3.
  37. Sashirekha S, Muthumary J (2007). Detection of taxol, an anticancer drug, from selected coelomycetous fungi. Indian J Sci Tech, 1, 1-0.
  38. Seenivasan A, Shubahgar S, Arvindan R, Viruthagiri T (2008). Microbial production and biomedical applications of lovastatin. Indian J Pharma Sci, 70, 701-9. https://doi.org/10.4103/0250-474X.49087
  39. Shi Y, Felley-Bosco E, Marti TM, Stahel RA (2012). Differential effects of lovastatin on cisplatin responses in normal human mesothelial cells versus cancer cells: implication for therapy. PloS one, 7, 453-54.
  40. Shibata MA, Ito Y, Morimoto J, Otsuki Y (2004). Lovastatin inhibits tumor growth and lung metastasis in mouse mammary carcinoma model: a p53-independent mitochondrialmediated apoptotic mechanism. Carcinogenesis, 25, 1887-8. https://doi.org/10.1093/carcin/bgh201
  41. Traverso N, Ricciarelli R, Nitti M, et al (2013). Role of glutathione in cancer progression and chemoresistance. Oxidative Med Cellular Longevity, 1-10
  42. Wong WWL, Clendening JW, Martirosyan A, et al (2007). Determinants of sensitivity to lovastatin-induced apoptosis in multiple myeloma. Molecular Cancer Therapeutics, 6, 1886-7. https://doi.org/10.1158/1535-7163.MCT-06-0745