Advertisement

Journal of Physiology and Biochemistry

, Volume 71, Issue 2, pp 191–204 | Cite as

d-pinitol mitigates tumor growth by modulating interleukins and hormones and induces apoptosis in rat breast carcinogenesis through inhibition of NF-κB

  • Thamaraiselvan Rengarajan
  • Natarajan Nandakumar
  • Peramaiyan Rajendran
  • Mohanraj Karthik Ganesh
  • Maruthaiveeran Periyasamy BalasubramanianEmail author
  • Ikuo Nishigaki
Original Paper

Abstract

Breast cancer is the most prevalent malignant neoplasm in the world, and chemoprevention through dietary intervention strategy is an emerging option to reduce the incidence. d-pinitol (DP), a major component of soya bean, possesses attractive biological actions. We have investigated whether d-pinitol have an effect on tumor growth in vivo against 7,12-dimethylbenz(a)anthracene (DMBA)-initiated rat mammary carcinogenesis and investigated its mechanism of action. Tumors were induced in Sprague–Dawley (SD) rats by a gastric dose of 20 mg/kg DMBA, and after 13 weeks of induction period, the rats were orally administered with d-pinitol for 45 days. At the end of the assay, animals in carcinogen control group prompted a tumor incidence of 100 % and developed a tumor volume of 8.35 ± 0.56, which was significantly reduced to 5.74 ± 0.32 for the animals treated with d-pinitol. The d-pinitol treatment not only decreased the tumor volume but also further examination revealed that tumors from animals that received d-pinitol reduced nuclear factor kappa B (NF-κB) activation which in turn results in modulation of its downstreaming p53 and proteins of caspase-3 family. Bcl-2 expression and caspase-3 activation were also decreased after d-pinitol supplementation leading to induction of apoptosis and finally cell death. Furthermore, the status of the inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin (IL)-2, IL-6, and tumor markers, lipid profile, and hormones was also significantly declined up on d-pinitol administration. Thus, it reveals the collective involvement of the abovementioned parameters along with NF-κB signaling through which d-pinitol induces apoptosis and subsequently suppresses breast cancer during DMBA-induced rat breast carcinogenesis.

Keywords

d-pinitol Apoptosis DMBA Breast cancer NF-κB Bcl-2 Caspase-3 p53 

Notes

Acknowledgments

The authors are extremely grateful to Dr. R. Venkatakrishna Murali, M.D., Ph.D., Professor and Head, Department of Pharmacology and Environmental Toxicology, Dr. A.L. Mudhaliar Post Graduate Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai—600113 for providing the laboratory facilities and the financial assistance from Department of Science and Technology (DST), New Delhi in the form of DST-PURSE-JRF is gratefully acknowledged and one of the authors “Natarajan Nandakumar” thank Prof. Jacob Gopas PhD, MPH (BGU, Israel) for his useful comments and encouragement. We thank Dr. S. Prakash, Ph.D., Assistant Professor, Department of Anatomy, Dr. ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai for interpreting the immunofluorescence micrographs and suggestions. We are also thankful to Mrs. Rita Rajan, CMC Vellore for transmission electron microscope studies and Dr. Sarasabharathy for interpretation.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Aggarwal BB, Shishodia S, Sandur SK, Pandey MK, Sethi G (2006) Inflammation and cancer: how hot is the link? Biochem Pharm 72(11):1605–1621CrossRefPubMedGoogle Scholar
  2. 2.
    Aggarwal BB, Takada Y (2005) Pro-apoptotic and anti-apoptotic effects of tumor necrosis factor in tumor cells. Role of nuclear transcription factor NF-kB. Cancer Treat Res 126:103–27CrossRefPubMedGoogle Scholar
  3. 3.
    Ahmed OI, Adel AM, Diab DR, Gobran NS (2006) Prognostic value of serum level of interleukin-6 and interleukin-8 in metastatic breast cancer patients. Egypt J Immunol 13:61–8PubMedGoogle Scholar
  4. 4.
    Alexander P (1975) Tumour immunology is perspective. In: Schcultz J, Leiff RC (eds) Critical factors in cancer immunology. Academic Press, New York, pp 213–222Google Scholar
  5. 5.
    Anbuselvam C, Vijayavel K, Balasubramanian MP (2007) Protective effect of Operculina turpethum against 7,12-dimethyl benz(a)anthracene induced oxidative stress with reference to breast cancer in experimental rats. Chem Biol Interacts 168:229–236CrossRefGoogle Scholar
  6. 6.
    Barros ACSD, Muranaka ENK, Mori LJ, Pelizon CHT, Iriya K, Giocondo K, Pinotti JA (2004) Induction of experimental mammary carcinogenesis in rats with 7,12-dimethylbenz(a) anthracene. Rev Hosp Clin Fac Med Sao Paulo 59(5):257–261CrossRefPubMedGoogle Scholar
  7. 7.
    Cagnol S, Mansour A, Van E, Schilling O, Chambard JC (2011) Raf-1 activation prevents caspase 9 processing downstream of apoptosome formation. J Signal Transduct 12–12Google Scholar
  8. 8.
    Chandy KG, Huebscher SG, Elias E, Berg J, Khan M, Burnett D (1983) Dual role of the liver in regulating polymeric IgA in man: studies on patients with liver diseases. Cltn Exp Immunol 52:207–218Google Scholar
  9. 9.
    Chatterton RT Jr, Geiger AS, Gann PH, Khan SA (2003) Formation of estrone and estradiol from estrone sulfate by normal breast parenchymal tissue. J Steroid Biochem Mol Biol 86:159–166CrossRefPubMedGoogle Scholar
  10. 10.
    Chaturvedi MM, Sung B, Yadav VR, Kannappan R, Aggarwal BB (2011) NF-kB addiction and its role in cancer: ‘one size does not fit all’. Oncogene 30:1615–1630CrossRefPubMedCentralPubMedGoogle Scholar
  11. 11.
    Chen Y, Chen L, Li JY, Mukaida N, Wang Q, Yang C (2011) ER and PEA3 co-activate IL-8 expression and promote the invasion of breast cancer cells. Cancer Biol Ther 11:497–511CrossRefPubMedGoogle Scholar
  12. 12.
    Chomczynski P, Sacchi N (1987) Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162(1):156–9CrossRefPubMedGoogle Scholar
  13. 13.
    Conney AH, Lou YR, Xie JG, Osawa T, Newmark HL, Liu Y, Chang RL, Huang MT (1997) Some perspectives on dietary inhibition of carcinogenesis: studies with curcumin and tea. Proc Soc Exp Biol Med 216:243–245CrossRefGoogle Scholar
  14. 14.
    Dabrosin C (2005) Increased extracellular local levels of estradiol in normal breast in vivo during the luteal phase of the menstrual cycle. J Endocrinol 187:103–108CrossRefPubMedGoogle Scholar
  15. 15.
    Damen J, Ramshorst JV, Hoeven RPV (1984) Alterations in plasma lipo protein and heparin releasable lipase activities in mice bearing the GRSL ascites tumor. Biochem Biophys Acta 793:287–296CrossRefPubMedGoogle Scholar
  16. 16.
    Dao JL, Bock FG, Crouch S (1959) Level of 3-methyl cholanthrene in mammary glands of rats after intragastric instillation of carcinogen. Proc Soc Exp Biol Med 102:635–638CrossRefPubMedGoogle Scholar
  17. 17.
    Eun Jung P, John MP (2002) Botanicals in cancer chemoprevention. Cancer Met Rev 21:231–255CrossRefGoogle Scholar
  18. 18.
    Fentiman IS (2001) Fixed and modifiable risk factors for breast cancer. Int J Clin Pract 55:527–530PubMedGoogle Scholar
  19. 19.
    Floch J, Less M, Stanley GHS (1951) A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226:497–509Google Scholar
  20. 20.
    Geren RI, Greenberg NH, MacDonald MM, Schumacher AM, Abbott BJ (1972) Protocols for screening chemicals agents and natural products against animal tumors and other biological systems. Cancer Chemother Rep 3:1–103Google Scholar
  21. 21.
    Guney N, Soydinc H, Basaran M, Bavbek S, Derin D, Camlica H, Yasasever V, Topuz E (2009) Serum levels of interleukin-6 and interleukin-10 in Turkish patients with aggressive non-Hodgkin’s lymphoma. APJCP 10:669–674PubMedGoogle Scholar
  22. 22.
    Harris L, Frische H, Mennel R (2007) American Society of Clinical Oncology 2007 update of recommendations for the use of tumor markers in breast cancer. Clinical practice guidelines of American Society of Clinical Oncology. J Clin Oncol 25:5287–5312CrossRefPubMedGoogle Scholar
  23. 23.
    Horn WT, Menahan LA (1981) A sensitive method for the determination of free fatty acids in plasma. J Lipid Res 23:377–381Google Scholar
  24. 24.
    Jose MM, Juan AS, Francisco JA, Javier M (2008) Intra cellular redox status and oxidative stress: implications for cell proliferartion, apoptosis and carcinogenesis. Arch Toxicol 82:273–299CrossRefGoogle Scholar
  25. 25.
    Kachoie P, Mohammad H, Pourgholami DL (2013) Morris inhibition of the IL-6 signaling pathway: a strategy to combat chronic inflammatory diseases and cancer. Cytokine Growth Factor Rev 24(2):163–73CrossRefGoogle Scholar
  26. 26.
    Katz (1983) Immunobiologic staging of patients with carcinoma of the hand and neck. Laryngoscope 93:445–463PubMedGoogle Scholar
  27. 27.
    Kozlowski L, Zakrzewska I, Tokajuk P, Wojtukiewicz MZ (2003) Concentration of interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-10 (IL-10) in blood serum of breast cancer patients. Rocz Akad Med Bialymst 48:82–4PubMedGoogle Scholar
  28. 28.
    Kumar K, Sachadanandam P, Arivazhagan R (1991) studies on the changes in plasma lipids and lipoproteins proteins in patients with benign and malignant breast cancer. Biochem Int 23:581–589PubMedGoogle Scholar
  29. 29.
    Kumar A, Takada Y, Boriek AM, Aggarwal BB (2004) Nuclear factor-kB: its role in health and disease. J Mol Med 82:434–48CrossRefPubMedGoogle Scholar
  30. 30.
    Kushlinskii NE, Orinovskii MB, Gurevich LE, Kazantseva IA, Talaeva Sh Z, Shirokii VP, Ermilova VD, Dvorova EK, Ozherelev AS (2004) Expression of biomolecular markers (Ki-67, PCNA, Bcl-2, BAX, BclX, and VEGF) in breast tumors. Bull Exp Biol Med 137:182–185CrossRefPubMedGoogle Scholar
  31. 31.
    Lamoureux G, Mandeville R, Poisson R (1982) Biologic markers and breast cancer. A nlulitparametric study increased serum protein levels. Cancer 49:502–512CrossRefPubMedGoogle Scholar
  32. 32.
    Leifeld L, Nattermann J, Fielenbach M, Schmitz V, Sauerbruch T, Spengler U (2006) Intrahepatic activation of caspases in human fulminant hepatic failure. Liver Int 26:872–879CrossRefPubMedGoogle Scholar
  33. 33.
    Lin TH, Tan TW, Tsai TH, Chen CC, Hsieh TF, Lee SS, Liu HH, Chen WC, Tang CH (2013) d-pinitol inhibits prostate cancer metastasis through inhibition of αVβ3 integrin by modulating FAK, c-Src and NF-κB pathways. Int J Mol Sci 14(5):9790–802CrossRefPubMedCentralPubMedGoogle Scholar
  34. 34.
    Liu FS, Jan YJ, Lai CR, Twu NF, Lu CH, Hung MJ, Hsieh YT, Chiou LC (2008) Expression analysis of apoptosis-related markers TP53, BCL-2, BAX and c-MYC in female genital tract sarcomas. J Chin Med Assoc 71:628–34CrossRefPubMedGoogle Scholar
  35. 35.
    Ludwig RL, Bates S, Vousden KH (1996) Differential activation of target cellular promoters by p53 mutants with impaired apoptotic function. Mol Cell Biol 16:49–52Google Scholar
  36. 36.
    Nandakumar N, Jayaprakash R, Balasubramanian MP (2011) Influence of hesperidin on renal cell surface glycoprotein content, nucleic acids, lysosomal enzymes and macromolecules against 7, 12-dimethylbenz [a] anthracene induced experimental breast carcinoma. JETO 2012 9(4):265–280Google Scholar
  37. 37.
    Nandhakumar R, Salini K, Niranjali Devaraj S (2012) Morin augments anticarcinogenic and antiproliferative efficacy against 7,12-dimethylbenz(a)-anthracene induced experimental mammary carcinogenesis. Mol Cell Biochem 364:79–92CrossRefPubMedGoogle Scholar
  38. 38.
    O’ Donovan N, Crown J, Stunell H, Hill A, McDermott E, O’Higgins N, Duffy M (2003) Caspase 3 in breast cancer. Clin Cancer Res 9:738–42Google Scholar
  39. 39.
    Parekh AC, Jung DH (1970) Cholesterol determinant with ferric chloride—uranyl acetate and sulphuric acid ferrous sulphate reagent. Anal Biochem 42:1423–1427Google Scholar
  40. 40.
    Parkin DM, Bray F, Ferlay J, Pisani P (2001) Estimating the world cancer burden: GLOBOCAN 2000. Int J Cancer 94:153CrossRefPubMedGoogle Scholar
  41. 41.
    Phillips DV, Dougherty DE, Smith AE (1982) Cyclitols in soybean. J Agric Food Chem 30:456–458CrossRefPubMedGoogle Scholar
  42. 42.
    Pizarro JG, Verdaguer E, Ancrenaz V, Junyent F, Sureda F, Pallas M, Folch J, Camins A (2011) Resveratrol inhibits proliferation and promotes apoptosis of neuroblastoma cells: role of sirtuin 1. Neurochem Res 36:187–194CrossRefPubMedGoogle Scholar
  43. 43.
    Premkumar V, Yuvaraj S, Vijayasarathy K, Gangadaran S, Sachdanandam P (2007) Serum cytokine levels of interleukin-1, -6, -8, tumour necrosis factor-α and vascular endothelial growth factor in breast cancer patients treated with tamoxifen and supplemented with co-enzyme Q10, riboflavin and niacin. Basic Clin Pharmacol Toxicol 100:387–391CrossRefPubMedGoogle Scholar
  44. 44.
    Pugalendhi P, Manoharan S, Baskaran N, Madhavan R, Nirmal (2010) Effects of genistein and daidzein, in combination, on the expression pattern of biomolecular markers (p53, PCNA, VEGF, iNOS, Bcl-2, and Bax) during 7,12-dimethylbenz(a)anthracene (DMBA) induced mammary carcinogenesis in Sprague-Dawley rats. Int J Biol Med Res 1(4):264–271Google Scholar
  45. 45.
    Reed JC (1995) Regulation of apoptosis by Bcl-2 family proteins and its role in cancer and chemoresistance. Curr Opin Oncol 7:541–6CrossRefPubMedGoogle Scholar
  46. 46.
    Rengarajan T, Nandakumar N, Rajendran P, Haribabu L, Nishigaki I, Balasubramanian MP (2014) d-pinitol promotes apoptosis in MCF-7 cells via induction of p53 and Bax and inhibition of Bcl-2 and NF-κB. Asian Pac J Cancer Prev 15(4):1757–62CrossRefPubMedGoogle Scholar
  47. 47.
    Rengarajan T, Nandakumar N, Balasubramanian MP (2012) d-pinitol a low-molecular cyclitol prevents 7,12-dimethylbenz [a] anthracene induced experimental breast cancer through regulating anti-apoptotic protein Bcl-2, mitochondrial and carbohydrate key metabolizing enzymes. Biomed Prevent Nutri 2(1):25–30CrossRefGoogle Scholar
  48. 48.
    Rice EW (1970) Triglycerides in serum. In: Standard method of clinical chemistry. Academic Press, New York 6: 213–222Google Scholar
  49. 49.
    Roberts MM, Bathgate EM, Stevenson A (1975) Serum immunoglobulin levels in patients with breast cancer. Cancer 36(1):221–224CrossRefPubMedGoogle Scholar
  50. 50.
    Rouser G, Fleisher S, Yamanoto A (1970) Two dimensional thin layer chromatographic separation of polar lipids and determination of phospholipids by phosphorous analysis of spots. Lipids 5:494–496CrossRefPubMedGoogle Scholar
  51. 51.
    Santer FR, Malinowska K, Culig Z, Cavarretta IT (2010) Interleukin-6 trans-signalling differentially regulates proliferation, migration, adhesion and maspin expression in human prostate cancer cells. Endo Relat Cancer 17(1):241–53CrossRefGoogle Scholar
  52. 52.
    Satpathy PK, Dutta KK, Mishra PR, Kar BC (1996) Glutaraldehyde coagulation test: standard curve and its applications to detect gammaglobulin level in kids. Ind Ved J 73:257–260Google Scholar
  53. 53.
    Sethi G, Ahn KS, Sung B, Bharat B, Aggarwal (2008) Pinitol targets nuclear factor-KB activation pathway leading to inhibition of gene products associated with proliferation, apoptosis, invasion, and angiogenesis. Mol Cancer Ther 7:1604–1614CrossRefPubMedGoogle Scholar
  54. 54.
    Shyamala G, Chou YC, Louie SG, Guzman RC, Smith GH, Nandi S (2002) Cellular expression of estrogen and progesterone receptors in mammary glands: regulation by hormones, development and aging. J Steroid Biochem Mol Biol 80(2):137–48CrossRefPubMedGoogle Scholar
  55. 55.
    Sierra A (2005) Metastases and their microenvironments: linking pathogenesis and therapy. Drug Resist Updat 8(4):247–57CrossRefPubMedGoogle Scholar
  56. 56.
    Sivakumar S, Palsamy P, Subramanian S (2010) Impact of d-pinitol on the attenuation of proinflammatory cytokines, hyperglycemia-mediated oxidative stress and protection of kidney tissue ultrastructure in streptozotocin-induced diabetic rats. Chem Biol Interact 188:237–245CrossRefPubMedGoogle Scholar
  57. 57.
    Stieber P, Nagel D, Ritzke C, Rossler N, Kirsch CM, Eiermann W, Fateh-Moghadam A (1992) Significance of bone alkaline phosphatase, CA 15-3 and CEA in the detection of bone metastases during the follow-up of patients suffering from breast carcinoma. Eur J Clin Chem Clin Biochem 30(12):809–14PubMedGoogle Scholar
  58. 58.
    Suchi K, Fujiwara H, Okamura S, Okamura H, Umehara S, Todo M (2011) Overexpression of interleukin-6 suppresses cisplatin-induced cytotoxicity in esophageal squamous cell carcinoma cells. Anticancer Res 31(1):67–75PubMedGoogle Scholar
  59. 59.
    Tennant B, Baldwin BHB, Raun RK, Norcross NL, Sandholm M (1979) Use of the glutaraldehyde coagulation test for the detection of hypogamma-globulinemia in neonatal calves. Am J Vet Med 174:848–853Google Scholar
  60. 60.
    Thiyagarajan R, Senthil Murugan K, Kavitha P, Anitha D, Prathiba S, Nagini (2012) Dietary chlorophyllin inhibits the canonical NF-kB signaling pathway and induces intrinsic apoptosis in a hamster model of oral oncogenesis. Food Chem Toxicol 50:867–876CrossRefPubMedGoogle Scholar
  61. 61.
    Uehara M, Kinoshita T, Hojo T, Akashi Tanaka S, Takashi E, Fukutomi (2008) Long-term prognostic study of carcinoembryonic antigen (CEA) and carbohydrate antigen 15-3 (CA 15-3) in breast cancer. Int J Clin Oncol 13:447–451CrossRefPubMedGoogle Scholar
  62. 62.
    Van Handel E (1961) Modification of the micro determination of triglycerides. Clin Chem 1:249–251Google Scholar
  63. 63.
    Vinothkumar V, Manoharan S, Sindhu G, Nirmal M, Vetrichelvi V (2012) Geraniol modulates cell proliferation, apoptosis, inflammation, and angiogenesis during 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis. Mol Cell Biochem 369:17–25CrossRefPubMedGoogle Scholar
  64. 64.
    Yager JD, Davidson NE (2006) Estrogen carcinogenesis in breast cancer. N Engl J Med 354(3):270–82CrossRefPubMedGoogle Scholar
  65. 65.
    Zuccari DAPC, Leonel C, Castroc R, Gelaleti GB, Jardim B, Moscheta M, Regiani V, Ferreira L, Lopes JR, de Santi Neto D, Estevesg JL (2012) An immunohistochemical study of interleukin-8 (IL-8) in breast cancer. Acta Histochem 114:571–576CrossRefPubMedGoogle Scholar

Copyright information

© University of Navarra 2015

Authors and Affiliations

  • Thamaraiselvan Rengarajan
    • 1
    • 2
  • Natarajan Nandakumar
    • 2
    • 4
  • Peramaiyan Rajendran
    • 3
  • Mohanraj Karthik Ganesh
    • 5
  • Maruthaiveeran Periyasamy Balasubramanian
    • 2
    Email author
  • Ikuo Nishigaki
    • 3
  1. 1.Department of Chemical Pathology, School of Medical SciencesUniversiti Sains MalaysiaKelantanMalaysia
  2. 2.Department of Pharmacology and Environmental Toxicology, Dr. ALM Post Graduate Institute of Basic Medical SciencesUniversity of MadrasChennaiIndia
  3. 3.NPO-International Laboratory of BiochemistryNagoyaJapan
  4. 4.The Shraga Segal Department of Microbiology, Immunology and GeneticsFaculty of Health Sciences, Ben Gurion University of the NegevBeer ShevaIsrael
  5. 5.Department of Anatomy, Dr. ALM PG IBMSUniversity of MadrasChennaiIndia

Personalised recommendations