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Pathology & Oncology Research

, Volume 25, Issue 1, pp 301–309 | Cite as

Invitro Evaluation of Torin2 and 2, 6-Dihydroxyacetophenone in Colorectal Cancer Therapy

  • Ankita Awasthi
  • Pharvendra Kumar
  • Chittur V. Srikanth
  • Shakti Sahi
  • Rekha PuriaEmail author
Original Article
  • 110 Downloads

Abstract

Colorectal cancer (CRC) is one of the most prevalent cancers diagnosed worldwide. Despite recent advances, resistance to cytotoxic and targeted therapy remains one of the greatest challenges in long-term management of colorectal cancer therapy. Recently established role of mTOR signaling in proliferation of CRC has incited for evaluation of mTOR kinase specific inhibitors in CRC therapy. Second generation mTOR kinase inhibitors including Torin2 has demonstrated efficient anticancer properties against variety of cancers and are in various stages of drug development. The time and financial constraints concomitant from discovery to development of efficient chemical inhibitors has redirected attention towards investigation of wide spread naturally occurring largely inexpensive compounds for their therapeutic potential. One such naturally occurring compound acetophenone derivative polyphenolic compound 2, 6-Dihydroxyacetophenone (DHAP) inhibits cell growth in different conditions. We investigated anticancer properties of both Torin2 and DHAP against colorectal cancer in HCT8 cell lines. Both Torin2 and DHAP inhibited growth of CRC cells at different concentrations by restricting multiple cellular functions e.g., cell cycle progression, cell migration and induced apoptosis. Treatment of HCT8 cells with natural compound DHAP resulted in reduced expression of mTOR pathway specific genes p70S6K1 and AKT1. In silico docking studies showed affinity of DHAP to mTOR kinase like Torin2. Taken together, our result vouches for role of Torin2 in CRC therapy and recommends DHAP an mTOR inhibitor, as a potential lead in the development of new therapeutic regimes against colorectal cancer.

Keywords

mTOR Kinase inhibitor Drug development Docking Natural Compound 

Notes

Acknowledgments

The authors are thankful to Dr. Vikrant Nain for critical reading of the manuscript and for providing valuable suggestions that increased scientific content and presentation of the manuscript. Financial assistance in the form of research grant from Department of Biotechnology, Ministry of Science &Technology, India to RP is duly acknowledged.

Funding

The work was supported by research grant to RP from Department of Biotechnology, Ministry of Science and Technology, Govt of India.

Compliance with Ethical Standards

Conflict of Interest

Authors declare no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    Lee C, Murray DW, Barber RT et al (1998) Siegel Et Al 1998.Pdf. Deep Res Part II Top Stud Oceanogr 45:2489–2501CrossRefGoogle Scholar
  2. 2.
    Society AC (2014) Colorectal Cancer Facts & Figures 2014-2016. Color Cancer Facts Fig :1–32.  https://doi.org/10.1101/gad.1593107
  3. 3.
    Guertin DA, Sabatini DM (2005) An expanding role for mTOR in cancer. Trends Mol Med 11:353–361.  https://doi.org/10.1016/j.molmed.2005.06.007 CrossRefGoogle Scholar
  4. 4.
    Liu Q, Thoreen C, Wang J et al (2009) MTOR mediated anti-cancer drug discovery. Drug Discov Today Ther Strateg 6:47–55.  https://doi.org/10.1016/j.ddstr.2009.12.001 CrossRefGoogle Scholar
  5. 5.
    Benjamin D, Colombi M, Moroni C, Hall MN (2011) Rapamycin passes the torch: a new generation of mTOR inhibitors. Nat Rev Drug Discov 10:868–880.  https://doi.org/10.1038/nrd3531 CrossRefGoogle Scholar
  6. 6.
    Manning G, Whyte DB, Martinez R et al (2002) The protein kinase complement of the human genome. Science 298:1912–1934.  https://doi.org/10.1126/science.1075762 CrossRefGoogle Scholar
  7. 7.
    LoPiccolo J, Blumenthal GM, Bernstein WB, Dennis PA (2008) Targeting the PI3K/Akt/mTOR pathway: Effective combinations and clinical considerations. Drug Resist Updat 11:32–50.  https://doi.org/10.1016/j.drup.2007.11.003 CrossRefGoogle Scholar
  8. 8.
    Efeyan A, Sabatini DM (2010) MTOR and cancer: Many loops in one pathway. Curr Opin Cell Biol 22:169–176.  https://doi.org/10.1016/j.ceb.2009.10.007 CrossRefGoogle Scholar
  9. 9.
    Murugan AK, Alzahrani A, Xing M (2013) Mutations in critical domains confer the human mTOR gene strong tumorigenicity. J Biol Chem 288:6511–6521.  https://doi.org/10.1074/jbc.M112.399485 CrossRefGoogle Scholar
  10. 10.
    Grabiner BC, Nardi V, Birsoy K et al (2014) A diverse array of cancer-associated MTOR mutations are hyperactivating and can predict rapamycin sensitivity. Cancer Discov 4:554–563.  https://doi.org/10.1158/2159-8290.CD-13-0929 CrossRefGoogle Scholar
  11. 11.
    Li W, Chang J, Wang S, et al (2015) miRNA-99b-5p suppresses liver metastasis of colorectal cancer by down-regulating mTOR. 6Google Scholar
  12. 12.
    Chresta CM, Davies BR, Hickson I et al (2010) AZD8055 is a potent, selective, and orally bioavailable ATP-competitive mammalian target of rapamycin kinase inhibitor with in vitro and in vivo antitumor activity. Cancer Res 70:288–298.  https://doi.org/10.1158/0008-5472.CAN-09-1751 CrossRefGoogle Scholar
  13. 13.
    Fujishita T, Aoki K, Lane HA et al (2008) Inhibition of the mTORC1 pathway suppresses intestinal polyp formation and reduces mortality in ApcDelta716 mice. Proc Natl Acad Sci U S A 105:13544–13549.  https://doi.org/10.1073/pnas.0800041105 CrossRefGoogle Scholar
  14. 14.
    Roulin D, Waselle L, Dormond-Meuwly A et al (2011) Targeting renal cell carcinoma with NVP-BEZ235, a dual PI3K/mTOR inhibitor, in combination with sorafenib. Mol Cancer 10:90.  https://doi.org/10.1186/1476-4598-10-90 CrossRefGoogle Scholar
  15. 15.
    Jiang BH, Liu LZ (2008) Role of mTOR in anticancer drug resistance: Perspectives for improved drug treatment. Drug Resist Updat 11:63–76.  https://doi.org/10.1016/j.drup.2008.03.001 CrossRefGoogle Scholar
  16. 16.
    Hammond WA, Swaika A, Mody K (2015) Pharmacologic resistance in colorectal cancer: a review. Ther Adv Med Oncol.  https://doi.org/10.1177/1758834015614530
  17. 17.
    Liu Q, Xu C, Kirubakaran S, Zhang X, Hur W, Liu Y, et al (2013) Characterization of Torin2, an ATP-competitive inhibitor of mTOR, ATM, and ATR. Cancer Res 73:2574–2586Google Scholar
  18. 18.
    Wang C, Wang X, Su Z, et al (2015) The novel mTOR inhibitor Torin-2 induces autophagy and downregulates the expression of UHRF1 to suppress hepatocarcinoma cell growth. 1708–1716.  https://doi.org/10.3892/or.2015.4146
  19. 19.
    Simioni C, Cani A, Martelli AM, et al Activity of the novel mTOR inhibitor Torin-2 in B-precursor acute lymphoblastic leukemia and its therapeutic potential to prevent Akt reactivation. 5Google Scholar
  20. 20.
    Ahmed M, Hussain AR, Bavi P et al (2014) High prevalence of mTOR complex activity can be targeted using Torin2 in papillary thyroid carcinoma. Carcinogenesis 35:1564–1572.  https://doi.org/10.1093/carcin/bgu051
  21. 21.
    Udayakumar D, Pandita RK, Horikoshi N et al (2016) Torin2 Suppresses Ionizing Radiation-Induced DNA Damage Repair. Radiat Res 538:527–538.  https://doi.org/10.1667/RR14373.1
  22. 22.
    Martínez-Luis S, Pérez-Vásquez A, Mata R (2007) Natural products with calmodulin inhibitor properties. Phytochemistry 68:1882–1903.  https://doi.org/10.1016/j.phytochem.2007.02.025 CrossRefGoogle Scholar
  23. 23.
    Badr CE, Van Hoppe S, Dumbuya H et al (2013) Targeting cancer cells with the natural compound obtusaquinone. J Natl Cancer Inst 105:643–653.  https://doi.org/10.1093/jnci/djt037 CrossRefGoogle Scholar
  24. 24.
    Sigurdsson S, Ögmundsdottir HM, Hallgrimsson J, Gudbjarnason S (2005) Antitumour activity of Angelica archangelica leaf extract. In: In Vivo (Brooklyn), vol 19, pp 191–194Google Scholar
  25. 25.
    Erdemgil FZ, Şanli S, Şanli N et al (2007) Determination of pKa values of some hydroxylated benzoic acids in methanol-water binary mixtures by LC methodology and potentiometry. Talanta 72:489–496.  https://doi.org/10.1016/j.talanta.2006.11.007 CrossRefGoogle Scholar
  26. 26.
    Nakamura ES, Kurosaki F, Arisawa M, Mukainaka T (2002) Cancer chemopreventive effects of constituents of Caesalpinia ferrea and related compounds. Cancer Lett 177:119–124Google Scholar
  27. 27.
    Pursh PC (2014) Polyphenols with Anti-Proliferative Activities from Penthorum chinense Pursh.:11045–11055.  https://doi.org/10.3390/molecules190811045
  28. 28.
    Bracht K, Nicholls AM, Liu Y, Bodmer WF (2010) 5-Fluorouracil response in a large panel of colorectal cancer cell lines is associated with mismatch repair deficiency. Brit J Cancer 103:340–346CrossRefGoogle Scholar
  29. 29.
    Miller JM (2003) GraphPad PRISM. Analysis 52:2–3. www.graphpad.com
  30. 30.
    Sadowski SM, Boufraqech M, Zhang L et al (2015) Torin2 targets dysregulated pathways in anaplastic thyroid cancer and inhibits tumor growth and metastasis. Oncotarget 6:18038–18049.  https://doi.org/10.18632/oncotarget.3833
  31. 31.
    Beck DE (2009) Colorectal cancer. Rev Gastroenterol Mex 8:2–3.  https://doi.org/10.1038/nrgastro.2011.208 Google Scholar
  32. 32.
    Liang C-C, Park AY, Guan J-L (2007) In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2:329–333.  https://doi.org/10.1038/nprot.2007.30 CrossRefGoogle Scholar
  33. 33.
    Evan GI, Vousden KH (2001) Proliferation, cell cycle and apoptosis in cancer. Nature 411:342–348.  https://doi.org/10.1038/35077213 CrossRefGoogle Scholar
  34. 34.
    Francipane MG, Lagasse E (2013) mTOR pathway in colorectal cancer: an update. Oncotarget 5:49–66.  https://doi.org/10.18632/oncotarget.1548 Google Scholar
  35. 35.
    Wang XW, Zhang YJ (2014) Targeting mTOR network in colorectal cancer therapy. World J Gastroenterol 20:4178–4188.  https://doi.org/10.3748/wjg.v20.i15.4178 CrossRefGoogle Scholar
  36. 36.
    Cai Z, Ke J, He X et al (2014) Significance of mTOR signaling and its inhibitor against cancer stem-like cells in colorectal cancer. Ann Surg Oncol 21:179–188.  https://doi.org/10.1245/s10434-013-3146-8 CrossRefGoogle Scholar
  37. 37.
    Passtoors WM, Beekman M, Deelen J et al (2013) Gene expression analysis of mTOR pathway: Association with human longevity. Aging Cell 12:24–31.  https://doi.org/10.1111/acel.12015 CrossRefGoogle Scholar
  38. 38.
    Riquelme I, Tapia O, Espinoza JA et al (2016) The Gene Expression Status of the PI3K/AKT/mTOR Pathway in Gastric Cancer Tissues and Cell Lines. Pathol Oncol Res.  https://doi.org/10.1007/s12253-016-0066-5
  39. 39.
    Wang N, Wu R, Cheng X et al (2014) New insights into mTOR structure and regulation. Chinese Sci Bull 59:2927–2935.  https://doi.org/10.1007/s11434-014-0417-0 CrossRefGoogle Scholar
  40. 40.
    Friesner RA, Banks JL, Murphy RB et al (2004) Glide: A New Approach for Rapid, Accurate Docking and Scoring. 1. Method and Assessment of Docking Accuracy. J Med Chem 47:1739–1749.  https://doi.org/10.1021/jm0306430 CrossRefGoogle Scholar
  41. 41.
    Friesner RA, Murphy RB, Repasky MP et al (2006) Extra precision glide: Docking and scoring incorporating a model of hydrophobic enclosure for protein-ligand complexes. J Med Chem 49:6177–6196.  https://doi.org/10.1021/jm051256o CrossRefGoogle Scholar
  42. 42.
    Link A, Balaguer F, Shen Y et al (2013) Curcumin Modulates DNA Methylation in Colorectal Cancer Cells. PLoS One.  https://doi.org/10.1371/journal.pone.0057709
  43. 43.
    Zhao X, Song H, Zuo Z et al (2013) Identification of miRNA-miRNA synergistic relationships in colorectal cancer. Int J Biol Macromol 55:98–103.  https://doi.org/10.1016/j.ijbiomac.2012.12.006 CrossRefGoogle Scholar
  44. 44.
    Blaser B, Waselle L, Dormond-Meuwly A et al (2012) Antitumor activities of ATP-competitive inhibitors of mTOR in colon cancer cells. BMC Cancer 12:86.  https://doi.org/10.1186/1471-2407-12-86 CrossRefGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2017

Authors and Affiliations

  1. 1.School of BiotechnologyGautam Buddha UniversityGreater NOIDAIndia
  2. 2.Regional Centre for BiotechnologyFaridabadIndia
  3. 3.Kalinga institute of industrial technologyKIIT UniversityBhubaneswarIndia

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