Tumor Biology

, Volume 37, Issue 2, pp 2083–2093 | Cite as

Gedunin abrogates aldose reductase, PI3K/Akt/mToR, and NF-κB signaling pathways to inhibit angiogenesis in a hamster model of oral carcinogenesis

  • Kranthi Kiran Kishore T
  • Raghu Ganugula
  • Deepak Reddy Gade
  • Geereddy Bhanuprakash Reddy
  • Siddavaram Nagini
Original Article


Aberrant activation of oncogenic signaling pathways plays a central role in tumor development and progression. The aim of this present study was to investigate the chemopreventive effects of the neem limonoid gedunin in the hamster model of oral cancer based on its ability to modulate aldose reductase (AR), phosphatidyl inositol-3-kinase (PI3K)/Akt, and nuclear factor kappa B (NF-κB) pathways to block angiogenesis. Administration of gedunin suppressed the development of HBP carcinomas by inhibiting PI3K/Akt and NF-κB pathways through the inactivation of Akt and inhibitory kappa B kinase (IKK), respectively. Immunoblot and molecular docking interactions revealed that inhibition of these signaling pathways may be mediated via inactivation of AR by gedunin. Gedunin blocked angiogenesis by downregulating the expression of miR-21 and the pro-angiogenic factors vascular endothelial growth factor and hypoxia inducible factor-1 alpha (HIF-1α). In conclusion, the results of the present study provide compelling evidence that gedunin prevents progression of hamster buccal pouch (HBP) carcinomas via inhibition of the kinases Akt, IKK, and AR, and the oncogenic transcription factors NF-κB and HIF-1α to block angiogenesis.


Gedunin Angiogenesis Aldose reductase Phosphatidyl inositol-3-kinase Akt Nuclear factor kappa B 



Financial support from the Indian Council of Medical Research, New Delhi, India, in the form of a Senior Research Fellowship to Mr. Kranthi Kiran Kishore T. is gratefully acknowledged.

Conflicts of interest


Authors’ contributions

SN, GBR, and TKK designed the experiments. TKK, GR, and GDR performed the experiments and analyzed the data. SN, TKK, and GBR were involved in the preparation of the manuscript.


  1. 1.
    Yedida GR, Nagini S, Mishra R. The importance of oncogenic transcription factors for oral cancer pathogenesis and treatment. Oral Surg Oral Med Oral Pathol Oral Radiol. 2013;116:179–88.CrossRefPubMedGoogle Scholar
  2. 2.
    Sever R, Brugge JS. Signal transduction in cancer. Cold Spring Harb Perspect Med. 2015;5:4.CrossRefGoogle Scholar
  3. 3.
    Davis WJ, Lehmann PZ, Li W. Nuclear PI3K signaling in cell growth and tumorigenesis. Front Cell Dev Biol. 2015;3:24.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Chaturvedi MM, Sung B, Yadav VR, Kannappan R, Aggarwal BB. NF-κB addiction and its role in cancer: ‘one size does not fit all’. Oncogene. 2011;30:1615–30.CrossRefPubMedGoogle Scholar
  5. 5.
    Porta C, Paglino C, Mosca A. Targeting PI3K/Akt/mTOR signaling in cancer. Front Oncol. 2014;4:64.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Simpson DR, Mell LK, Cohen EE. Targeting the PI3K/AKT/mTOR pathway in squamous cell carcinoma of the head and neck. Oral Oncol. 2015;51:291–8.CrossRefPubMedGoogle Scholar
  7. 7.
    Li W, Wang H, Kuang CY, Zhu JK, Yu Y, Qin ZX, et al. An essential role for the Id1/PI3K/Akt/NFκB/survivin signalling pathway in promoting the proliferation endothelial progenitor cells in vitro. Mol Cell Biochem. 2012;363:135–45.CrossRefPubMedGoogle Scholar
  8. 8.
    Sun ZJ, Chen G, Hu X, Zhang W, Liu Y, Zhu LX, et al. Activation of PI3K/Akt/IKK-alpha/NF-kappaB signalling pathway is required for the apoptosis-evasion in human salivary adenoid cystic carcinoma: its inhibition by quercetin. Apoptosis. 2010;15:850–63.CrossRefPubMedGoogle Scholar
  9. 9.
    Bai D, Ueno L, Vogt PK. Akt-mediated regulation of NFkappaB and the essentialness of NFkappaB for the oncogenicity of PI3K and Akt. Int J Cancer. 2009;125:2863–70.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Shen HM, Tergaonkar V. NFkappaB signaling in carcinogenesis and as a potential molecular target for cancer therapy. Apoptosis. 2009;14:348–63.CrossRefPubMedGoogle Scholar
  11. 11.
    Hussain AR, Ahmed SO, Ahmed M, Khan OS, Al Abdulmohsen S, Platanias LC, et al. Cross-talk between NF-κB and the PI3-kinase/AKT pathway can be targeted in primary effusion lymphoma (PEL) cell lines for efficient apoptosis. PLoS One. 2012;7:e39945.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Tammali R, Reddy AB, Srivastava SK, Ramana KV. Inhibition of aldose reductase prevents angiogenesis in vitro and in vivo. Angiogenesis. 2011;14:209–21.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Tammali R, Saxena A, Srivastava SK, Ramana KV. Aldose reductase inhibition prevents hypoxia-induced increase in hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) by regulating 26 S proteasome-mediated protein degradation in human colon cancer cells. J Biol Chem. 2011;286:24089–100.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Sicard F, Gayral M, Lulka H, Buscail L, Cordelier P. Targeting miR-21 for the therapy of pancreatic cancer. Mol Ther. 2013;21:986–94.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Biswas K, Chattopadhyay I, Banerjee RK. Biological activities and medicinal properties of neem (Azadirachta indica). Curr Sci. 2002;82:1336–45.Google Scholar
  16. 16.
    Subapriya R, Nagini S. Medicinal properties of neem leaves: a review. Curr Med Chem Anticancer Agents. 2005;5:149–56.CrossRefPubMedGoogle Scholar
  17. 17.
    Harish Kumar G, Chandra Mohan KV, Jagannadha Rao A, Nagini S. Nimbolide, a limonoid from Azadirachta indica inhibits proliferation and induces apoptosis of human choriocarcinoma (BeWo) cells. Invest New Drugs. 2009;27:246–52.CrossRefPubMedGoogle Scholar
  18. 18.
    Kavitha K, Vidya Priyadarsini R, Anitha P, Ramalingam K, Sakthivel R, Purushothaman G, et al. Nimbolide, a neem limonoid abrogates canonical NF-κB and Wnt signaling to induce caspase-dependent apoptosis in human hepatocarcinoma (HepG2) cells. Eur J Pharmacol. 2012;681:6–14.CrossRefPubMedGoogle Scholar
  19. 19.
    Vidya Priyadarsini R, Manikandan P, Harish Kumar G, Nagini S. The neem limonoids azadirachtin and nimbolide inhibit hamster cheek pouch carcinogenesis by modulating xenobiotic-metabolizing enzymes, DNA damage, antioxidants, invasion, and angiogenesis. Free Radic Res. 2009;43:492–504.CrossRefGoogle Scholar
  20. 20.
    Harish Kumar G, Vidya Priyadarsini R, Vinothini G, Vidjaya Letchoumy P, Nagini S. The neem limonoids azadirachtin and nimbolide inhibit cell proliferation and induce apoptosis in an animal model of oral oncogenesis. Invest New Drugs. 2010;28:392–401.CrossRefPubMedGoogle Scholar
  21. 21.
    Priyadarsini RV, Murugan RS, Sripriya P, Karunagaran D, Nagini S. The neem limonoids azadirachtin and nimbolide induce cell cycle arrest and mitochondria-mediated apoptosis in human cervical cancer (HeLa) cells. Free Radic Res. 2010;44:624–34.CrossRefPubMedGoogle Scholar
  22. 22.
    Kavitha K, Kranthi Kiran Kishore T, Bhatnagar RS, Nagini S. Cytomodulin-1, a synthetic peptide abrogates oncogenic signaling pathways to impede invasion and angiogenesis in the hamster cheek pouch carcinogenesis model. Biochimie. 2014;102:56–67.CrossRefPubMedGoogle Scholar
  23. 23.
    Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–9.CrossRefPubMedGoogle Scholar
  24. 24.
    Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem. 1976;72:248–54.CrossRefPubMedGoogle Scholar
  25. 25.
    Hieronymus H, Lamb J, Ross KN, Peng XP, Clement C, Rodina A, et al. Gene expression signature-based chemical genomic prediction identifies a novel class of HSP90 pathway modulators. Cancer Cell. 2006;10:321–30.CrossRefPubMedGoogle Scholar
  26. 26.
    Uddin SJ, Nahar L, Shilpi JA, Shoeb M, Borkowski T, Gibbons S, et al. Gedunin, a limonoid from Xylocarpus granatum, inhibits the growth of CaCo-2 colon cancer cell line in vitro. Phytother Res. 2007;21:757–61.CrossRefPubMedGoogle Scholar
  27. 27.
    Kamath SG, Chen N, Xiong Y, Wenham R, Apte S, Humphrey M, et al. Gedunin, a novel natural substance, inhibits ovarian cancer cell proliferation. Int J Gynecol Cancer. 2009;19:1564–9.CrossRefPubMedGoogle Scholar
  28. 28.
    Patwardhan CA, Fauq A, Peterson LB, Miller GC, Blagg BS, Chadli A. Gedunin inactivates the cochaperone p23 protein causing cancer cell death by apoptosis. J Biol Chem. 2013;288:7313–25.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Mabuchi S, Kuroda H, Takahashi R, Sasano T. The PI3K/AKT/mTOR pathway as a therapeutic target in ovarian cancer. Gynecol Oncol. 2015;137:173–9.CrossRefPubMedGoogle Scholar
  30. 30.
    Pal I, Mandal M. PI3K and Akt as molecular targets for cancer therapy: current clinical outcomes. Acta Pharmacol Sin. 2012;33:1441–58.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Kowshik J, Giri H, Kishore TK, Kesavan R, Vankudavath RN, Reddy GB, et al. Ellagic acid inhibits VEGF/VEGFR2, PI3K/Akt and MAPK signaling cascades in the hamster cheek pouch carcinogenesis model. Anticancer Agents Med Chem. 2014;14:1249–60.CrossRefPubMedGoogle Scholar
  32. 32.
    Strickland LR, Pal HC, Elmets CA, Afaq F. Targeting drivers of melanoma with synthetic small molecules and phytochemicals. Cancer Lett. 2015;359:20–35.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Hinz M, Scheidereit C. The IκB kinase complex in NF-κB regulation and beyond. EMBO Rep. 2014;15:46–61.CrossRefPubMedGoogle Scholar
  34. 34.
    Saxena A, Tammali R, Ramana KV, Srivastava SK. Aldose reductase inhibition prevents colon cancer growth by restoring phosphatase and tensin homolog through modulation of miR-21 and FOXO3a. Antioxid Redox Signal. 2013;18:11249–62.CrossRefGoogle Scholar
  35. 35.
    Saxena A, Shoeb M, Ramana KV, Srivastava SK. Aldose reductase inhibition suppresses colon cancer cell viability by modulating microRNA-21 mediated programmed cell death 4 (PDCD4) expression. Eur J Cancer. 2013;49:3311–9.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Zhang W, Bai W, Zhang W. MiR-21 suppresses the anticancer activities of curcumin by targeting PTEN gene in human non-small cell lung cancer A549 cells. Clin Transl Oncol. 2014;16:708–13.CrossRefPubMedGoogle Scholar
  37. 37.
    Muthenna P, Suryanarayana P, Gunda SK, Petrash JM JM, Reddy GB. Inhibition of aldose reductase by dietary antioxidant curcumin: mechanism of inhibition, specificity and significance. FEBS Lett. 2009;583:3637–42.CrossRefPubMedGoogle Scholar
  38. 38.
    Wang G, Dai F, Yu K, Jia Z, Zhang A, Huang Q, et al. Resveratrol inhibits glioma cell growth via targeting oncogenic microRNAs and multiple signaling pathway. Int J Oncol. 2015;46:1739–47.PubMedGoogle Scholar
  39. 39.
    Zhou C, Ding J, Wu Y. Resveratrol induces apoptosis of bladder cancer cells via miR-21 regulation of the Akt/Bcl-2 signaling pathway. Mol Med Rep. 2014;9:1467–73.PubMedGoogle Scholar
  40. 40.
    Shin VY, Jin H, Ng EK, Cheng AS, Chong WW, Wong CY, et al. NF-κB targets miR-16 and miR-21 in gastric cancer: involvement of prostaglandin E receptors. Carcinogenesis. 2011;32:240–5.CrossRefPubMedGoogle Scholar
  41. 41.
    Yadav UC, Srivastava SK, Ramana KV. Prevention of VEGF-induced growth and tube formation in human retinal endothelial cells by aldose reductase inhibition. J Diabetes Complications. 2012;26:369–77.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Liu LZ, Li C, Chen Q, Jing Y, Carpenter R, Jiang Y, et al. MiR-21 induced angiogenesis through AKT and ERK activation and HIF-1α expression. PLoS One. 2011;6:e19139.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Kranthi Kiran Kishore T
    • 1
  • Raghu Ganugula
    • 2
  • Deepak Reddy Gade
    • 3
  • Geereddy Bhanuprakash Reddy
    • 2
  • Siddavaram Nagini
    • 1
  1. 1.Department of Biochemistry and Biotechnology, Faculty of ScienceAnnamalai UniversityAnnamalainagarIndia
  2. 2.Biochemistry DivisionNational Institute of NutritionHyderabadIndia
  3. 3.Medicinal Chemistry Research DivisionVishnu Institute of Pharmaceutical Education and ResearchNarsapurIndia

Personalised recommendations