Advertisement

Tetrandrine isolated from Cyclea peltata induces cytotoxicity and apoptosis through ROS and caspase pathways in breast and pancreatic cancer cells

  • Bhagya N.
  • K. R. ChandrashekarEmail author
  • Ashwini Prabhu
  • P. D. Rekha
Article
  • 66 Downloads

Abstract

Tetrandrine is a bisbenzylisoquinoline alkaloid known to exhibit anticancer activity against different cancers. In the present study, the cytotoxic effect of tetrandrine isolated from Cyclea peltata on pancreatic (PANC-1) and breast (MDA-MB-231) cancer cells was evaluated in vitro with an attempt to understand the role of tetrandrine on the generation of reactive oxygen species (ROS) and caspase activation. Results demonstrate the dose- and time-dependant cytotoxic effect of tetradrine on both MDA-MB-231 and PANC-1 cells with IC50 values ranging between 51 and 54 μM and 22 and 27 μM for 24 h and 48 h of incubation respectively. In addition, treatment of MDA-MB-231 and PANC-1 cells with tetrandrine showed the shrunken cytoplasm and damaged cell membrane in a dose- and time-dependant manner under the microscope. Also, tetrandrine treatment revealed an elevated levels of reactive oxygen species and increased activities of caspase-8, -9 and -3 confirming the apoptosis of cells through both extrinsic death receptor and intrinsic caspase activation. Therefore, the present study suggests the apoptosis of cells with the activation of caspase pathways mainly intrinsic pathway as a downstream event of tetrandrine-induced ROS generation. Hence, reactive oxygen species-mediated caspase activation pathway may be potentially targeted with the use of tetrandrine to treat breast and pancreatic cancers.

Keywords

Tetrandrine Cancer ROS Caspase Apoptosis 

Notes

Acknowledgments

Senior author was supported by University Grants Commission, New Delhi, India [No. F.15-1/2012-13/PDFWM-2012-13-GE-KER-13766(SA-II)]. Authors also acknowledge Board of Research in Nuclear Sciences (BRNS), Mumbai, India (No. 35/14/40/2014-BRNS/1118) and Department of Science and technology (DST), India (EMR/2016/001734), for bearing a part of consumable cost.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interests.

Supplementary material

11626_2019_332_MOESM1_ESM.jpg (65 kb)
Figure S1 LC-ESI-MS spectrum of Tet B showing m/z value of 622.8. HPLC chromatogram of the isolated tetrandrine is showed inside the mass spectrum towards left side and chemical structure of tetrandrine is given inside the mass spectrum towards right side. (JPG 64 kb)

References

  1. Azad MB, Chen Y, Gibson SB (2009) Regulation of autophagy by reactive oxygen species (ROS): implications for cancer progression and treatment. Antioxid Redox Signal 11:777–790CrossRefGoogle Scholar
  2. Baehrecke EH (2005) Autophagy: dual roles in life and death? Nat Rev Mol Cell Biol 6:505–510CrossRefGoogle Scholar
  3. Baig S, Seevasant I, Mohamad J, Mukheem A, Huri HZ, Kamarul T (2016) Potential of apoptotic pathway-targeted cancer therapeutic research: where do we stand? Cell Death and Disease 7:e2058.  https://doi.org/10.1038/cddis.2015.275 CrossRefGoogle Scholar
  4. Bhagya N, Chandrashekar KR (2016) Tetrandrine—a molecule of wide bioactivity. Phytochemistry 125:5–13CrossRefGoogle Scholar
  5. Bhagya N, Chandrashekar KR (2017) A process for the extraction and purification of tetrandrine. Indian patent; 201741016069. Publication date: 16/06/2017Google Scholar
  6. Bhagya N, Chandrashekar KR (2018) Tetrandrine and cancer—an overview on the molecular approach. Biomed Pharmacother 97:624–632CrossRefGoogle Scholar
  7. Bhagya N, Chandrashekar KR (2019) Optimization of column chromatography technique for the isolation of tetrandrine from Cyclea peltata and LC-ESI-MS based quantification and validation of the method. Nat Prod Research.  https://doi.org/10.1080/14786419.2018.1503660
  8. Bianchini G, Balko JM, Mayer IA, Sanders ME, Gianni L (2016) Triple-negative breast cancer: challenges and opportunities of a heterogeneous disease. Nat Rev Clin Oncol 13:674–690CrossRefGoogle Scholar
  9. Campani D, Esposito I, Boggi U, Cecchetti D, Menicagli M, De Negri F, Colizzi L, Del Chiaro M, Mosca F, Fornaciari G, Bevilacqua G (2001) Bcl-2 expression in pancreas development and pancreatic cancer progression. J Pathol 194:444–450CrossRefGoogle Scholar
  10. Cande C, Cecconi F, Dessen P, Kroemer G (2002) Apoptosis-inducing factor (AIF): key to the conserved caspase-independent pathways of cell death? J Cell Sci 15:4727–4734CrossRefGoogle Scholar
  11. Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB (1987) Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res 47(4):936–942Google Scholar
  12. Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35:495–516CrossRefGoogle Scholar
  13. Fulda S, Debatin K-M (2006) Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 25:4798–4811CrossRefGoogle Scholar
  14. Gamble JS (1958) Flora of Presidency of Madras, vol I-III. Sri Gouranga Press Pvt. Ltd, CalcuttaGoogle Scholar
  15. Holohan C, Schaeybroeck SV, Longley DB, Johnston PG (2013) Cancer drug resistance: an evolving paradigm. Nat Rev Cancer 13:714–726CrossRefGoogle Scholar
  16. Huang AC, Lien JC, Lin MW, Yang JS, Wu PP, Chang SJ, Lai TY (2013) Tetrandrine induces cell death in SAS human oral cancer cells through caspase activation-dependent apoptosis and LC3-I and LC3-II activation-dependent autophagy. Int J Oncol 43:485–494CrossRefGoogle Scholar
  17. Kidd JF, Pilkington MF, Schell MJ, Fogarty KE, Skepper JN, Taylor CW, Thorn P (2002) Paclitaxel affects cytosolic calcium signals by opening the mitochondrial permeability transition pore. J Biol Chem 2:6504–6510CrossRefGoogle Scholar
  18. Kleeff J, Korc M, Apte M, La Vecchia C, Johnson CD, Biankin AV, Neale RE, Tempero M, Tuveson DA, Hruban RH, Neoptolemos JP (2016) Pancreatic cancer. Nat Rev Dis Primers 21; 2:16022.  https://doi.org/10.1038/nrdp.2016.22 CrossRefGoogle Scholar
  19. Kondo Y, Kanzawa T, Sawaya R, Kondo S (2005) The role of autophagy in cancer development and response to therapy. Nat Rev Cancer 5:726–734CrossRefGoogle Scholar
  20. Li X, Su B, Liu R, Wu D, He D (2011) Tetrandrine induces apoptosis and triggers caspase cascade in human bladder cancer cells. J Surg Res 166:e45–e51CrossRefGoogle Scholar
  21. Ling L-U, Tan K-B, Lin H, Chiu GNC (2011) The role of reactive oxygen species and autophagy in safingol-induced cell death. Cell Death Dis 2:e129CrossRefGoogle Scholar
  22. Liu C, Ke G, Mao X, Li W (2011) Tetrandrine induces apoptosis by activating reactive oxygen speies and repressing Akt activity in human hepatocellular carcinoma. Int J Cancer 129:1519–1531CrossRefGoogle Scholar
  23. Meena J, Santhy KS (2015a) Antitumor activity of methanolic extract of Cyclea peltata. Int J Phytomed 7:185–192Google Scholar
  24. Meena J, Santhy KS (2015b) Efficacy of methanolic extract of Cyclea peltata as a potent anticancer equivalent. Eur J Environ Ecol 2:65–71Google Scholar
  25. Qin R, Shen H, Cao Y, Fang Y, Li H, Chen Q, Xu W (2013) Tetrandrine induces mitochondria-mediated apoptosis in human gastric cancer BGC-823 cells. PLoS One 8:1–10Google Scholar
  26. Qiu W, Su M, Xie F, Ai J, Ren Y, Zhang J, Guan R, He W, Gong Y, Guo (2014) Tetrandrine blocks autophagic flux and induces apoptosis via energetic impairment in cancer cells. Cell Death Dis 5:e1123.  https://doi.org/10.1038/cddis.2014.84 CrossRefGoogle Scholar
  27. Riedl SJ, Shi Y (2004) Molecular mechanisms of caspase regulation during apoptosis. Nat Rev Mol Cell Biol 5:897–907.  https://doi.org/10.1038/nrm1496 CrossRefGoogle Scholar
  28. Robertson JD, Gogvadze V, Zhivotovsky B, Orrenius S (2000) Distinct pathways for stimulation of cytochrome c release by etoposide. J Biol Chem 275:32438–32443CrossRefGoogle Scholar
  29. Saelens X, Festjens N, Walle LV, van Gurp M, van Loo G, Vandenabeele P (2004) Toxic proteins released from mitochondria in cell death. Oncogene 23:2861–2874CrossRefGoogle Scholar
  30. Singh K, Dong Q, Koul S, Koul HK (2016a) Tetrandrine induces cell cycle arrest at G0/G1 boundary and cell death in vitro pancreatic cancer cells. Biochem Mol Biol Abstract Number 638:1Google Scholar
  31. Singh K, Shanmugam PST, Koul S, Dong Q, Koul N, Koul HK (2016b) Abstract 3516: Tetrandrine promotes pancreatic cancer cell apoptosis in vitro and tumor regression in vivo. Molecular and cellular biology, genetics. Proceedings: AACR 107th annual meeting April 16-20, New Orleans, LA 2016  https://doi.org/10.1158/1538-7445.AM2016-3516
  32. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65:87–108CrossRefGoogle Scholar
  33. Wang H, Liu T, Li L, Wang Q, Yu C, Liu X, Li W (2015) Tetrandrine is a potent cell autophagy agonist via activated intracellular reactive oxygen species. Cell Biosci 5.  https://doi.org/10.1186/2045-3701-5-4
  34. Wu D, Yotnda P (2011) Production and detection of reactive oxygen species (ROS) in cancers. J Vis Exp 57, pii: 3357.  https://doi.org/10.3791/3357
  35. Yu F-S, Yu C-S, Chen J-C, Yang J-L, Lu H-F, Chang S-J, Lin M-W, Chung J-G (2016) Tetrandrine induces apoptosis via caspase-8, -9, and -3 and poly (ADP ribose) polymerase dependent pathways and autophagy through Beclin-1/ LC3-I, II signaling pathways in human oral cancer HSC-3 cells. Environ Toxicol 31:395–406CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Bhagya N.
    • 1
  • K. R. Chandrashekar
    • 1
    Email author
  • Ashwini Prabhu
    • 2
  • P. D. Rekha
    • 2
  1. 1.Department of Applied BotanyMangalore UniversityMangaloreIndia
  2. 2.Yenepoya Research CentreYenepoya (Deemed to be University)MangaloreIndia

Personalised recommendations