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Pathology & Oncology Research

, Volume 19, Issue 4, pp 763–770 | Cite as

Naringenin (Citrus Flavonone) Induces Growth Inhibition, Cell Cycle Arrest and Apoptosis in Human Hepatocellular Carcinoma Cells

  • Duraikannu ArulEmail author
  • Perumal Subramanian
Research

Abstract

Search for new substances with antiproliferative activity and apoptosis inducing potential towards HepG2 cells is important since HCC is notoriously resistant to conventional chemotherapy. Dietary phytochemicals with significant anti-proliferative and apoptosis inducing potential are considered as agents promising for cancer therapy. Naringenin, a common dietary flavonoid abundantly present in fruits and vegetables, is believed to possess strong cytotoxic activity in numerous types of cancer cells. However, the detailed molecular mechanisms of its antiproliferative effects and apoptosis induction are still unclear. In this study, we investigated antiproliferative and apoptosis-inducing effect of naringenin in human hepatocellular carcinoma HepG2 cells. Naringenin was shown to inhibit the proliferation of HepG2 cells resulted partly from an accumulation of cells in the G0/G1 and G2/M phase of the cell cycle. Naringenin induced a rapid accumulation of p53, which might account for the naringenin-induced G0/G1 and G2/M phase arrests in Hep G2 cells. In addition, naringenin have been shown to induce apoptosis as evidenced by nuclei damage and increased proportion of apoptotic cells detected by flow cytometry analysis. Naringenin triggered the mitochondrial-mediated apoptosis pathway as shown by an increased ratio of Bax/Bcl-2, subsequent release of cytochrome C, and sequential activation of caspase-3. Our results showed that naringenin had inhibitory effect on the growth of HepG2 cell line through inhibition of cell proliferation and apoptosis induction. The elucidation of the drug targets of naringenin on inhibition of tumor cells growth should enable further development of naringenin for liver cancer therapy.

Keywords

Naringenin Hepatocellular carcinoma cells Cell proliferation Apoptosis 

Notes

Acknowledgements

We wish to thank Prof. C.M. Elinos-Báez, Universidad Nacional Autónoma de México, Mexico for her gift of antibodies against Bcl-2 and Bax; Prof. S. Soddu, Regina Elena Cancer Institute, Italy for her kind gift of anti p53; Prof. Haider Raza, UAE University, UAE for her kind gift of antibodies against caspase-3; Prof. Ronald Jemmerson, Department of Microbiology, Northwestern University for her kind gift of the antibody against cytochrome C.

Conflict of interest

The authors declare that there are no conflicts of interest.

References

  1. 1.
    Semela D, Heim M (2011) Hepatocellular carcinoma. Ther Umsch 68:213–217PubMedCrossRefGoogle Scholar
  2. 2.
    Wysocki PJ (2010) Targeted therapy of hepatocellular cancer. Expert Opin Investig Drugs 19:265–274PubMedCrossRefGoogle Scholar
  3. 3.
    Zender L, Kubicka S (2008) Molecular pathogenesis and targeted therapy of hepatocellular carcinoma. Onkologie 31:550–5PubMedCrossRefGoogle Scholar
  4. 4.
    Ye SL, Takayama T, Geschwind J, Marrero JA, Bronowicki JP (2010) Current approaches to the treatment of early hepatocellular carcinoma. Oncologist 15:34–41PubMedCrossRefGoogle Scholar
  5. 5.
    Zhang ZM, Guo JX, Zhang ZC, Jiang N, Zhang ZY, Pan LJ (2011) Therapeutic options for intermediate-advanced hepatocellular carcinoma. World J Gastroenterol 17:1685–1689PubMedCrossRefGoogle Scholar
  6. 6.
    Tan AC, Konczak I, Sze DM, Ramzan I (2011) Molecular pathways for cancer chemoprevention by dietary phytochemicals. Nutr Cancer 63:495–505PubMedCrossRefGoogle Scholar
  7. 7.
    Díaz-Laviada I (2010) Effect of capsaicin on prostate cancer cells. Future Oncol 6:1545–1550PubMedCrossRefGoogle Scholar
  8. 8.
    Lee KW, Bode AM, Dong Z (2011) Molecular targets of phytochemicals for cancer prevention. Nat Rev Cancer 11:211–218PubMedCrossRefGoogle Scholar
  9. 9.
    Harapu CD, Miron A, Cuciureanu M, Cuciureanu R (2010) Flavonoids-bioactive compounds in fruits juice. Rev Med Chir Soc Med Nat Iasi 114:1209–1214PubMedGoogle Scholar
  10. 10.
    Wahle KW, Brown I, Rotondo D, Heys SD (2011) Plant phenolics in the prevention and treatment of cancer. Adv Exp Med Biol 698:36–51CrossRefGoogle Scholar
  11. 11.
    Clere N, Faure S, Martinez MC, Andriantsitohaina R (2011) Anticancer properties of flavonoids: Roles in various stages of carcinogenesis. Cardiovasc Hematol Agents Med Chem 9:62–77PubMedCrossRefGoogle Scholar
  12. 12.
    Renugadevi J, Prabu SM (2010) Cadmium-induced hepatotoxicity in rats and the protective effect of naringenin. Exp Toxicol Pathol 62:171–181PubMedCrossRefGoogle Scholar
  13. 13.
    Sabarinathan D, Mahalakshmi P, Vanisree AJ (2011) Naringenin, a flavanone inhibits the proliferation of cerebrally implanted C6 glioma cells in rats. Chem Biol Interact 189:26–36PubMedCrossRefGoogle Scholar
  14. 14.
    Ekambaram G, Rajendran P, Magesh V, Sakthisekaran D (2008) Naringenin reduces tumor size and weight lost in N-methyl-N’-nitro-N-nitrosoguanidine-induced gastric carcinogenesis in rats. Nutr Res 28:106–112PubMedCrossRefGoogle Scholar
  15. 15.
    Qin L, Jin L, Lu L, Lu X, Zhang C, Zhang F, Liang W (2011) Naringenin reduces lung metastasis in a breast cancer resection model. Protein Cell 2:507–516PubMedCrossRefGoogle Scholar
  16. 16.
    Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, Van Oers MH (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415–1420PubMedGoogle Scholar
  17. 17.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  18. 18.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  19. 19.
    Knowles LM, Zigrossi DA, Tauber RA, Hightower C, Milner JA (2000) Flavonoids suppress androgen-independent human prostate tumor proliferation. Nutr Cancer 38:116–122PubMedCrossRefGoogle Scholar
  20. 20.
    Wang H, Li YL, She WZ, Guo GQ, Jiang ZY, Cen YZ, Fan ZY (2007) Studies on antiproliferative effect of flavones compounds isolated from Yao herb medicines. Zhong Yao Cai 30:980–984PubMedGoogle Scholar
  21. 21.
    Lin E, Lin WH, Wang SY, Chen CS, Liao JW, Chang HW, Chen SC, Lin KY, Wang L, Yang HL, Hseu YC (2012) Flavokawain B inhibits growth of human squamous carcinoma cells: Involvement of apoptosis and cell cycle dysregulation in vitro and in vivo. J Nutr Biochem 23:368–378PubMedCrossRefGoogle Scholar
  22. 22.
    Zhong Z, Chen X, Tan W, Xu Z, Zhou K, Wu T, Cui L, Wang Y (2011) Germacrone inhibits the proliferation of breast cancer cell lines by inducing cell cycle arrest and promoting apoptosis. Eur J Pharmacol 667:50–55PubMedCrossRefGoogle Scholar
  23. 23.
    Suzuki K, Matsubara H (2011) Recent advances in p53 research and cancer treatment. J Biomed Biotechnol 2011:978312. doi: 10.1155/2011/978312 PubMedGoogle Scholar
  24. 24.
    Giono LE, Manfredi JJ (2006) The p53 tumor suppressor participates in multiple cell cycle checkpoints. J Cell Physiol 209:13–20PubMedCrossRefGoogle Scholar
  25. 25.
    Taylor WR, Stark GR (2001) Regulation of the G2/M transition by p53. Oncogene 20:1803–1815PubMedCrossRefGoogle Scholar
  26. 26.
    Alshatwi AA, Shafi G, Hasan TN, Al-Hazzani AA, Alsaif MA, Alfawaz MA, Lei KY, Munshi A (2011) Apoptosis-mediated inhibition of human breast cancer cell proliferation by lemon citrus extract. Asian Pac J Cancer Prev 12:1555–1559PubMedGoogle Scholar
  27. 27.
    Park HS, Kim GY, Nam TJ, Deuk Kim N, Hyun Choi Y (2011) Antiproliferative activity of fucoidan was associated with the induction of apoptosis and autophagy in AGS human gastric cancer cells. J Food Sci 26:77–83CrossRefGoogle Scholar
  28. 28.
    Tan AC, Konczak I, Ramzan I, Sze DM (2011) Native Australian fruit polyphenols inhibit cell viability and induce apoptosis in human cancer cell lines. Nutr Cancer 63:444–55PubMedCrossRefGoogle Scholar
  29. 29.
    Chidambara Murthy KN, Jayaprakasha GK, Kumar V, Rathore KS, Patil BS (2011) Citrus limonin and its glucoside inhibit colon adenocarcinoma cell proliferation through apoptosis. J Agric Food Chem 59:2314–2323PubMedCrossRefGoogle Scholar
  30. 30.
    Plati J, Bucur O, Khosravi-Far R (2011) Apoptotic cell signaling in cancer progression and therapy. Integr Biol (Camb) 4:279–296CrossRefGoogle Scholar
  31. 31.
    Kelly PN, Strasser A (2011) The role of Bcl-2 and its pro-survival relatives in tumourigenesis and cancer therapy. Cell Death Differ 18:1414–1424PubMedCrossRefGoogle Scholar
  32. 32.
    Martinou JC, Youle RJ (2011) Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Dev Cell 21:92–101PubMedCrossRefGoogle Scholar
  33. 33.
    Wang G, Zhan Y, Wang H, Li W (2012) ABT-263 sensitizes TRAIL-resistant hepatocarcinoma cells by downregulating the Bcl-2 family of anti-apoptotic protein. Can Chemother Pharmacol 69:799–805CrossRefGoogle Scholar
  34. 34.
    Jin CY, Park C, Lee JH, Chung KT, Kwon TK, Kim GY, Choi BT, Choi YH (2009) Naringenin-induced apoptosis is attenuated by Bcl-2 but restored by the small molecule Bcl-2 inhibitor, HA 14–1, in human leukemia U937 cells. Toxicol In Vitro 23:259–65PubMedCrossRefGoogle Scholar
  35. 35.
    Park JH, Jin CY, Lee BK, Kim GY, Choi YH, Jeong YK (2008) Naringenin induces apoptosis through downregulation of Akt and caspase-3 activation in human leukemia THP-1 cells. Food Chem Toxicol 46:3684–3690PubMedCrossRefGoogle Scholar
  36. 36.
    Lee ER, Kang YJ, Choi HJ, Kang GH, Kim JH, Kim BW, Han YS, Nah SY, Paik HD, Park YS (2007) Cho SG (2007) induction of apoptotic cell death by synthetic naringenin derivatives in human lung epithelial carcinoma A549 cells. Biol Pharm Bull 30:2394–8PubMedCrossRefGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2013

Authors and Affiliations

  1. 1.Department of Biochemistry and BiotechnologyFaculty of Science, Annamalai UniversityTamil NaduIndia

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