A polysaccharide from Tinospora cordifolia stem induces cell cycle arrest in human breast cancer cell lines MCF-7 and MDA-MB-231
Keywords:Anticancer, Breast Cancer, Cell Cycle, Phytotherapy, ADME/T.
AbstractPlant-based therapies are practiced for various human and veterinary ailments since time immemorial. The present study is concerned with finding the anti-breast cancer potential of a novel polysaccharide isolated from the methanolic extract of Tinospora cordifolia stem. The compound tested on MCF-7 and MDA-MB-231 proved to have potential to induce death of both the cell lines with IC50 at 100 Î¼M, as revealed in MTT, and LDH assays, and AO/EtBr staining. DNA fragmentation studies indicated damage to DNA. Flow cytometric analysis showed polysaccharide-induced cell cycle arrest at G2/M phase in both the cell lines. Western blot studies made it evident that the polysaccharide inhibits cell cycle progression via change in the expression of cell cycle regulators such as Cyclin D1, Cyclin D3 and p18 INK4. In the in-silico approach the structure of the compound was drawn using ChemSketch and the ADME/T properties of the compound were analyzed using Accord for Excel software. The compound possesses good ADME/T properties required for an active drug. The compound was found to possess anticancer efficacy via its effect on cell cycle regulatory proteins in the breast cancer cell lines and also satisfied the ADME/T properties of a drug. Hence, the novel compound isolated from T. cordifolia stem may be evaluated further so as to develop it as a breast cancer drug.
Murthy NS, Agarwal UK, Chaudhry K, Saxena S. A study on time trends in incidence of breast cancer – Indian scenario. European Journal of Cancer Care. 2007. 16:185–6. https://doi.org/10.1111/j.1365-2354.2006.00761.x PMid:17371429
Hortobagyi GN, De la Garza Salazar J, et al. The global breast cancer burden: variations in epidemiology and survival. Clinical Breast Cancer. 2005. 6: 391–401. https://doi.org/10.3816/CBC.2005.n.043 PMid:16381622
Sherr CJ. Mammalian G1 cyclins. Cell. 1993. 73:1059–65. https://doi.org/10.1016/0092-8674(93)90636-5
Hirai H, Roussel MF, Kato JY, Ashmun RA, Sherr CJ. Novel INK4 proteins, p19 and p18, are specific inhibitors of the cyclin d-dependent kinases CDK4 and CDK6. Molecular and Cellular Biology. 1995. 15: 2672–81. https://doi.org/10.1128/MCB.15.5.2672 PMid:7739547 PMCid:PMC230497
Sherr CJ, Roberts JM. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes and Development. 1999. 13:1501–12. https://doi.org/10.1101/ gad.13.12.1501 PMid:10385618
El-Diery WS, Harper JW, O'Connor PM, et al. WAF1/ CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Research. 1994. 54: 1169–74.
Toyoshima H, Hunter T. p27, a novel inhibitor of G1 cyclinCdk protein kinase activity, is related to p21. Cell. 1994. 78: 67–74. https://doi.org/10.1016/0092-8674(94)90573–8
Sa G, Stacey DW. p27 expression is regulated by separate signalling pathways, downstream of Ras, in each cell cycle phase. Experimental Cell Research. 2004. 300: 427–39. https://doi.org/10.1016/j.yexcr.2004.07.032 PMid:15475007
Weinert T, Lydall D. Cell cycle checkpoints, genetic instability and cancer. Seminars in Cancer Biology. 1993. 4: 129–40. PMid:8513148
Rao EV, Rao MV. Studies on the polysaccharide preparation (Guduchi Satwa) derived from Tinospora cordifolia. Indian Journal of Pharmaceutical Sciences. 1981. 43:103–6.
Singh N, Singh SM, Shrivastava P. Effect of Tinospora cordifolia on the antitumor activity of tumor-associated macrophagesderived dendritic cells. Immunopharmacology and Immunotoxicology. 2005. 27:1–14. https://doi.org/10.1081/IPH-51287 https://doi.org/10.1081/IPH200051287 PMid:15803856
Sangeetha MK, Raghavendran Balaji HR, Gayathri V, Vasanthi HR. Tinospora cordifolia attenuates oxidative stress and distorted carbohydrate metabolism in experimentally induced type 2 diabetes in rats. Journal of Natural Medicines. 2011. 65:544–50 https://doi.org/10.1007/s11418-011-0538-6 PMid:21538233
Ali H, Dixit S. Extraction optimization of Tinospora cordifolia and assessment of the anticancer activity of its alkaloid palmatine. Scientific World Journal. 2013. 3762–16. doi:10.1155/2013/376216 https://doi.org/10.1155/2013/376216
Leyon PV, Kuttan G. Inhibitory effect of a polysaccharide from Tinospora cordifolia on experimental metastasis. Journal of Ethnopharmacology. 2004. 90:233–7 https://doi.org/10.1016/j.jep.2003.09.046 PMid:15013186
Chaudhary R, Jahan S, Goyal PK. Chemopreventive potential of an Indian medicinal plant (Tinospora cordifolia) on skin carcinogenesis in mice. Journal of Environmental Pathology, Toxicology and Oncology. 2008. 27:233–43 https://doi.org/10.1615/JEnvironPatholToxicolOncol.v27.i3.70 PMid:18652570
Dhanasekaran M, Baskar AA, Ignacimuthu S, Agastian P, Duraipandiyan V. Chemopreventive potential of epoxy clerodane diterpene from Tinospora cordifolia against diethylnitrosamine-induced hepatocellular carcinoma. Investigational New Drugs. 2009. 27:347–55. https://doi.org/10.1007/s10637-008-9181-9 PMid:18853103
Thippeswamy G, Salimath BP. Induction of caspase-3 activated DNase mediated apoptosis by hexane fraction of Tinospora cordifolia in EAT cells. Environmental Toxicology and Pharmacology. 2007. 23:212–20. https://doi.org/10.1016/j.etap.2006.10.004 PMid:21783760
Maliyakkal N, Udupa N, Pai KSR, Rangarajan A. Cytotoxic and apoptotic activities of extracts of Withania somnifera and Tinospora cordifolia in human breast cancer cells. International Journal of Applied Research in Natural Products. 2013. 6:1–10.
Sharma P, Parmar J, Sharma P, Verma P, Goyal PK. Radiation-induced testicular injury and its amelioration by Tinospora cordifolia (An Indian medicinal plant) extract. Evidence Based Complementary and Alternative Medicine. 2011. Article ID 643847.
Rajalakshmi M, Eliza J, Priya CE, Nirmala A, Daisy P. Antidiabetic properties of Tinospora cordifolia stem extracts on streptozotocin- induced diabetic rats. African Journal of Pharmacy and Pharmacology. 2009. 3: 171–80
Manikkam R, Roy A. Î²-cell regenerative efficacy of a polysaccharide isolated from methanolic extract of Tinospora cordifolia stem on streptozotocin-induced diabetic Wistar rats. Chemico Biological Interactions. 2016. 243: 45–53 https://doi.org/10.1016/j.cbi.2015.11.021 PMid:26616445
Lowry OH, Rosebrough NJ, Farr AL, Randall R (1951).Protein Measurement with the Folin Phenol reagent. Journal of Biological Chemistry, 193: 265–275. PMid:14907713
Sawadogo WR, Schumacher M, Teiten MH, Dicato M, Diederich M. Traditional pharmacopoeia, plants and derived compounds for cancer therapy. Biochemical Pharmacology. 2012. 84:1225–40. https://doi.org/10.1016/j.bcp.2012.07.021 PMid:22846603
Duffy R, Wade C, Chang R. Discovery of anticancer drugs from antimalarial natural products: a MEDLINE literature review. Drug Discovery Today. 2012. 17:942–53 https://doi.org/10.1016/j.drudis.2012.03.013 PMid:22504324
Johnson DE, Wolfgang GH. Predicting human safety: screening and computational approaches. Drug Discovery Today. 2000. 5:445–54 https://doi.org/10.1016/S1359-6446(00)01559-2
Cotter TG. Apoptosis and cancer: the genesis of a research field. Nature Review: Cancer. 2009. 9:501–7 https://doi.org/10.1038/nrc2663 PMid:19550425
Molders P, Hogberg J, Orrenirs S. Methods in Enzymology. Vol. 52, Eds. Fleischer S., Packer L., Academic Press, New York, pp. 60–71. 1978.
Spector DL, Goldman RD, Leinwand LA. Cell: a Laboratory Manual, Vol. 1, Chap. 15, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp. 1–24. 1998.
Compton MM. A biochemical hallmark of apoptosis: internucleosomal degradation of the genome. Cancer Metastasis Review. 1992. 11:105–19 https://doi.org/10.1007/BF00048058 PMid:1327565
Williams GH, Stoeber K. The cell cycle and cancer. Journal of Pathology. 2012. 226: 352–64 https://doi.org/10.1002/path.3022 PMid:21990031
Stewart ZA, Westfall MD, Pietenpol JA. Cell-cycle dysregulation and anticancer therapy. Trends in Pharmacological Sciences. 2003. 24:139–45. https://doi.org/10.1016/S01656147(03)00026-9
Russell A, Thompson MA, Hendley J, Trute L, Armes J, Germain D. Cyclin D1 and D3 associate with the SCF complex and are coordinately elevated in breast cancer. Oncogene. 1999. 18: 1983–91. https://doi.org/10.1038/sj.onc.1202511 PMid:10208420
Wang C, Li Z, Fu M, Bouras T, Pestell RG. Signal transduction mediated by cyclin D1: from mitogens to cell proliferation: a molecular target with therapeutic potential. Cancer Treatment and Research. 2004. 119: 217–237. https://doi.org/10.1007/1-4020-7847-1_11 PMid:15164880
Franklin DS, Godfrey VL, Lee H, Kovalev GI, et al. CDK inhibitors p18INK4c and p27Kip1 mediate two separate pathways to collaboratively suppress pituitary tumorigenesis. Genes and Development. 1998. 12: 2899–911. https://doi.org/10.1101/gad.12.18.2899 PMid:9744866 PMCid:PMC317173