Effect of a Proprietary Commiphora mukul Gum Resin Extract and Medium-Chain Triglyceride Preparation (GU-MCT810) on hypoxia-inducible factor-1 pathway in HepG2 cell line

Authors

  • Cheppail Ramachandran Chief Science Officer and Sr. Research Scientist, Nicklaus Children's Hospital Miami Childrens Health System Miami, FL 33155, USA United States.
  • Gilda M. Portalatin Dharma Biomedical LLC, Miami, FL 33156 United States
  • Karl-W Quirin Flavex Naturextrakte, GmbH, Rehlingen, Germany.
  • Enrique Escalon Miami Children's Hospital, Miami, FL 33155 United States.
  • Steven J. Melnick Miami Children's Hospital, Miami, FL 33155 Dharma Biomedical LLC, Miami, FL 33156 United States.

Keywords:

Commiphora, hypoxia-inducible factor-1, HepG2 cell

Abstract

The heterodimeric transcription factor HIF-1 is responsible for the regulation of genes that facilitate adaptation and survival of cells under hypoxic conditions. HIF-1 gene expression is also associated with angiogenesis, glucose transport, nitric oxide synthase activity and cell proliferation through the regulation of hundreds of genes associated with HIF-1 pathway. GU-MCT810 is a nutraceutical ingredient complex that includes a Commiphora mukul (guggul) extract prepared by a supercritical CO2-co-solvent extraction with ethanol and medium chain triglyceride (MCT) oil composed of C8 and C10 fatty acids. Since cancer cells use glycolytic pathway, 2-deoxyglucose (2-DG) has been reported to inhibit the glycolysis. We have investigated the anticancer potential of GU-MCT810 with and without 2-DG in HepG2 human hepatoma cell line. Even though GU-MCT810 and 2-DG are individually weakly cytotoxic, the combination is synergistic with combination index (CI) values of 0.21, 0.22 and 0.88 at IC50, IC75 and IC90 levels, respectively. The combination also showed a synergistic inhibitory effect on ATP-synthesis in HepG2 cells. GU-MCT810 inhibits CoCl2-induced HIF-1α expression significantly in a dose-dependent manner with complete inhibition at 50 µg/ml concentration. GU-MCT810 upregulates Bax and p21 genes and down regulates Bcl-2, BNIP3 and mutant p53 genes associated with apoptosis. It also down regulates proteins associated angiogenesis (VEGF, VEGF-R), cell proliferation (IGF-2), glucose transport (GLUT1) and adaptogenesis (HSP70 and HSP90). These results indicate that GU-MCT810 can be combined with 2-DG for inhibition of HIF-1 pathway genes which would be useful for elimination of refractory cancer cells present in the hypoxic region of human tumors.

References

. Wang GL, Jiang BH, Rue EA, Semenza GL. Hypoxia-inducible factor 1 is a basic-helix-loop-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci USA 1995;92:5510-5514.

. Semenza GL. Hypoxia-incibel factor 1: master regulator of O2 homeostasis. Curr Opin Genet Dev 1998;8:588-594.

. Carmeliet P, Dor Y, Herbet JM, Fukumura D, Brusselmans K, Dewerchin M, et al. Role of HIF-1α in hypoxia-mediated apoptosis, cell proliferation and tumor angiogenesis. Nature 1998;394:485-490.

. Laderoute KR, Amin K, Calaoagan JM, Knapp M, Le T, Orduna J, et al. 5’-AMP –activated protein kinase (AMPK) is induced by low-oxygen and glucose deprivation conditions found in solid-tumor microenvironments. Mol Cell Biol 2006;26:5336-5347.

. Semenza GL. Involvement of hypoxia-inducible factor 1 in human cancer. Intern Med 2002;41: 79-83.

. Shi Y, Fang WG. Hypoxia-inducible factor-1 in tumor angiogenesis. World J Gastroenterol 2004;10:1082-1087.

. Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer 2003;3:721-732.

. Vaupel P. The role of hypoxia-Induced factors in tumor progression. Oncologist 2004;9:10-17.

. Zagzag D, Zhong H, Scalzitti JM, Laughner E, Simons JW, Semenza GL. Expression of hypoxia-inducible factor 1α in brain tumors: association with angiogenesis, invasion and progression. Cancer 2000;88:2606-2618.

. Goldberg MA, Dunning SP, Bunn HF. Regulation of the erythropoietin gene: evidence that the oxygen sensor is a heme protein. Science (Washington DC) 1988;242:1412-1415.

. Yao SY, Soutto M, Sriram S. Preconditioning with cobalt chloride or desferrioxamine protects oligodendrocyte cell line (MO3.13) from tumor necrosis factor-alpha-mediated cell death. J Nuerosci Res 2008;86:2403-2413.

. Bellozerov VE, Van Meir EG. Hypoxia inducible factor-1: a novel target for cancer therapy. Anti-Cancer Drugs 2005;16:901-909.

. Rapisarda A, Uranchimeg B, Sordet O, Pomier Y, Shoemaker RH, Melillo G. Topoisomerase I-mediated inhibition of hypoxia-inducible factor 1: mechanism and therapeutic implications. Cancer Res 2004;64:1475-1482.

. Ramachandran C, Nair SM, Quirin K-W, Melnick SJ. Hypolipidemic effects of a proprietary Commiphora mukul gum resin extract and medium-chain triglyceride preparation (GU-MCT). J Evid Based Complement Altern Med 2013;8:248-256.

. Ramachandran C, Lollett IV, Escalon E, Quirin K-W, Melnick SJ. Anticancer potential and mechanism of mango ginger (Curcuma amada Roxb.) supercritical CO2 extract in human glioblastoma cells. J. Evid Based Complement Altern Med 2014; 20:109-119.

. Ramachandran C, Quirin K-W, Esalon E, Lollett IV and Melnick SJ. Therapeutic effect of supercritical CO2 extracts of Curcuma species with cancer drugs in rhabdomyosarcoma cell lines. Phytother Res 2015;29:1152-1160..

. Ramachandran C, Resek AP, Escalon E, Aviram A, Melnick SJ. Potentiation of gemcitabine by Turmeric Force in pancreatic cancer cell lines. Oncol Rep 2010; 23:1529-1535.

. Chou TC, Talalay P. Analysis of combined drug effects: a new look at an old problem. Trends Pharmacol 1983;4:450-453.

. Ramachandran C, Nair SM, Escalon E, Aviram A, Melnick SJ. Potentiation of etoposide and temozolomide cytotoxicity by curcumin and Turmeric ForceTM in brain tumor cell lines. J Complement Integr Med 2012; Article20. DOI:10.1515/1553-3840.1614.

. Zhong H, De Marzo AM, Laughner E, Lim M, Hilton DA, Zagzag D, et al. Overexpression of hypoxia-inducible factor 1alpha in common human cancers and their metastases. Cancer Res 1999;59:5830-5835.

. Talks KL, Turley H, Gatter KC, Maxwell, PH, Pugh CW, Ratcliffe PJ, et al. The expression and distribution of the hypoxia-inducible factors HIF-1alpha and HIF-2alpha in normal human tissues, cancers, and tumor-associated macrophages. Am J Pathol 2000;157:411-421.

. Ryan HE, Poloni M, McNulty W, Elson D, Gassmann M, Arbeit JM, et al. Hypoxia-inducible factor-1alpha is a positive factor in solid tumor growth. Cancer Res 2000;60:4010-4015.

. Chen K, Zhao S, Nakada K, Kuge Y, Tamaki N, Okada F, et al. Dominant-negative hypoxia-inducible factor-1alpha reduces tumorigenicity of pancreatic cancer cells through the suppression of glucose metabolism. Am J Pathol 2003;162:1283-1291.

. Rapisarda A, Uranchimeg B, Scudiero DA, Selby M, Sausville EA, Shoemaker RH, et al. Identification of small molecule inhibitors of hypoxia-inducible factor 1 transcriptional activation pathway. Cancer Res 2002;62:4316-4324.

. Pelicano H, Martin DS, Xu RH, Huang P. Glycolysis inhibition for anticancer treatment. Oncogene 2006; 25:4633-4646.

. Hersey P, Watts R, Zhang XD, Jackett J. Metabolic approaches to treatment of melanoma. Clin Cancer Res 2009;15:6490-6494.

. Brown J. Effects of 2-deoxyglucose on carbohydrate metabolism: review of the literature and studies in the rat. Metabolism 1962;11:1098-1112.

. Weindruch R, Keenan KP, Carney JM, Fernandes G, Feuers RJ, Floyd RA, et al. Caloric restriction mimetics: metabolic interventions. J Gerontol A Biol Sci Med Sci 2001;1:20-33.

. Little E, Ramakrishnan M, Roy B, Gazit G, Lee S. The glucose-regulated proteins (GRP78 and GRP94): functions, gene regulation, and applications. Crit Rev Eukaryot Gene Expr 1994;4:1-18.

. Kang HT, Hwang ES. 2-Deoxyglucose: an anticancer and antiviral therapeutic, but not any more a low glucose mimetic. Life Sci 2000;78:1392-1399.

. Liu H, Hu YP, Savaraj N, Priebe W, Lampidis TJ. Hypersensitization of tumor cells to glycolytic inhibitors. Biochemistry 2001;40:5542-5547.

. Lampidis TJ, Kurtoglu M, Maher JC, Liu H, Krishan A, Sheft V, et al. Efficacy of 2-halogen substituted D-glucose analogs in blocking glycolysis and killing “hypoxic tumor cells”. Cancer Chemother Pharmacol 2006;58:725-734.

. Maschek G, Savaraj M, Priebe W, Braunschweiger P, Hamilton K, Tidmarsh GF, et al. 2-deoxy-D-glucose increases the efficacy of adriamycin and paclitaxel in human osteosarcoma and non-small cell lung cancers in vivo. Cancer Res 2004;64:31-34.

. Liu H, Jiang CC, Lavi CJ, Croft A, Dong L, Tseng H-Y, et al. 3-Deoxy-D-glucose enhances TRAIL-induced apoptosis in human melanoma cells through XBP-1-mediated up-regulation of TRAIL-R2. Mol Cancer 2009; 8:122-139.

. Zhong D, Xiong L, Liu T, Liu X, Chen J, Sun SY, et al. The glycolytic inhibitor 2-deoxyglucose activates multiple prosurvival pathways through IGF 1R. J Biol Chem 2009;284:23225-23233.

. Maher JC, Krishan A, Lampidis TJ. Cancer Chemother Pharmacol 2004; 53:116–122.

. Semsenza GL, Roth PH, Fang HM, Wang GL. Transcriptional regulation of genes encoding glycolytic enzymes by hypoxia-inducible factor 1. J Biol Chem 1994;269:23757-23763.

. Shimoda LA, Fallon M, Pisarcik S, Wang J, Semenza GL. HIF-1 regulates hypoxic induction of NHE1 expression and alkalinization of intracellular pH in pulmonary arterial myocytes. Am J Physiol Lung Cell Mol Physiol 2006;29:L941-949.

. Fukuda R, Zhang H, Kim J, Shimoda L, Dang CV, Semenza GL. HIF-1 regulates cytochrome oxidase subunits to optimize efficiency of respiration in hypoxic cells. Cell 2007;129:111-112,

. Nizet V, Johnson RS. Interdependence of hypoxic and innate immune responses. Nat Rev Immunol 2009;9:609-617.

. Ke Q, Costa M. Hypoxia-inducible factor-1(HIF-1). Mol Pharmacol 2006;70:1469-1680.

. Wang GL, Jiang BH, Rue EA, Semenza GL. Desferrioxamine induces erythropoietin gene expression and hypoxia-inducible factor 1 DNA-binding activity: implications for models of hypoxia signal transduction. Blood 1995;82:3610-3615.

. Wiener CM, Booth G, Semenza GL. In vivo expression of mRNAs encoding hypoxia-inducible factor 1. Biochem Biophys Res Commun 1996;225:485–488.

. Yu AY, Frid MG, Shimoda LA, Wiener CM, Stenmark K, Semenza GL. Temporal,

spatial, and oxygen-regulated expression of hypoxia-inducible factor-1 in the lung. Am J Physiol 1998;275:L818–826.

. Zagorska A, Dulak J. HIF-1: the knowns and unknowns of hypoxia sensing. Acta Biochim Pol 2004;51:563-85.

. Lichtinghagen R, Musholt PB, Lein M, Romer A, Rudolph B, Kristiansen G, et al. Different mRNA and protein expression of matrix metalloproteinases 2 and 9 and tissue inhibitor of metalloproteinase 1 in benign and malignant prostate tissue. Eur Urol 2002;2:398-406.

. Sarro SM, Unruh TL, Zuccolo J, Sanyal R, Lider JM, Auer-Grzesiak IA, et al. Quantification of CD20 mRNA and protein levels in chronic lymphocytic leukemia suggests a post-trnascriptional defect. Leuk Res 2010;34:1670-3.

. Stark AM, Pfannesnschmidt S, Tscheslog H, Maass N, Rosel F, Mehdorn HM,et al. Reduced mRNA and protein expression of BCL-2 versus decreased mRNA and increased protein expression of BAX in breast cancer brain metastases: a real-time PCR and immunohistochemical evaluation. Neurol Res 2006;28:787-793.

. Shebl FM, Pinto LA, Garia-Pineres A, Lempicki R, Williams M, Harro C, et al. Comparison of mRNA and protein measures of cytokines following vaccination with human papillomavirus-16 L1 virus particles. Cancer Epidemiol Biomarkers Prev 2010;19:978-981.

Downloads

Published

30-09-2015

How to Cite

1.
Cheppail Ramachandran, Gilda M. Portalatin, Karl-W Quirin, Enrique Escalon, Steven J. Melnick. Effect of a Proprietary Commiphora mukul Gum Resin Extract and Medium-Chain Triglyceride Preparation (GU-MCT810) on hypoxia-inducible factor-1 pathway in HepG2 cell line. ijp [Internet]. 2015 Sep. 30 [cited 2024 Nov. 2];7(3):324-36. Available from: https://ijp.arjournals.org/index.php/ijp/article/view/421

Issue

Section

Original Research Articles