High Glucose-reduced Apoptosis in Human Breast Cancer Cells Is Mediated by Activation of NF-κB

Keywords: Aerobic glycolysis, High glucose concentration, NF-Κb, Warburg effect


Tumor cells rely on glycolysis for their energy supply with the production of lactate even in normoxia condition, which is named aerobic glycolysis or Warburg effect. Therefore, high glucose (HG) concentration provides a favorable condition for increasing proliferation, angiogenesis and decreasing apoptosis, but its molecular mechanisms are still unknown. The objective of this study is to investigate HG condition on tumor cells behavior including proliferation, apoptosis, and an angiogenesis mediator. In this study, MCF-7 derived from human breast adenocarcinoma, were cultured in DMEM with two different concentrations of glucose for 48 h (5.5 mM as normal glucose (NG) condition and 25 mM as HG condition). We used Zingiber officinale extraction for the inhibition of NF-κB. Cell proliferation assay was done by direct counting, cell viability by MTT method, bcl-2 by Immunocytochemistry, apoptosis by Hoechst/PI double staining and vascular endothelium growth factor (VEGF) by ELISA. Results showed that HG increased lactate production, significantly. HG increased cell proliferation, cell viability, VEGF secretion, and bcl-2 expression while it decreased apoptosis. However, when HG was combined with Zingiber officinale extraction, cell proliferation, cell viability, VEGF secretion and bcl-2 expression decreased and apoptosis increased significantly due to inhibition of NF-κB. Results revealed that HG increased cell proliferation, angiogenesis and decreased apoptosis due to activation of NF-κB pathway. Moreover, the probable mechanism of the activation of NF-κB in HG is increasing reactive oxygen species (ROS) in this condition that can activate NF-κB directly.


1. Gupta C, Tikoo K. High glucose and insulin differentially modulates proliferation in MCF-7 and MDA-MB-231 cells. Journal Molucular Endocrinology. 2013;51(1):119-29.

2. Wahdan-Alaswad R, Fan Z, Edgerton SM, Liu B, Deng X-S, Arnadottir SS, et al. Glucose promotes breast cancer aggression and reduces metformin efficacy. Cell cycle. 2013;12(24):3759-69.

3. Kim J-w, Dang CV. Cancer's molecular sweet tooth and the Warburg effect. Cancer research. 2006;66(18):8927-30.

4. Villarreal-Garza C, Shaw-Dulin R, Lara-Medina F, Bacon L, Rivera D, Urzua L, et al. Impact of diabetes and hyperglycemia on survival in advanced breast cancer patients. Experimental diabetes research. 2012;2012.

5. Zeng L, Biernacka KM, Holly JM, Jarrett C, Morrison AA, Morgan A, et al. Hyperglycaemia confers resistance to chemotherapy on breast cancer cells: the role of fatty acid synthase. Endocrine-related cancer. 2010;17(2):539-51.

6. Xu R-h, Pelicano H, Zhou Y, Carew JS, Feng L, Bhalla KN, et al. Inhibition of glycolysis in cancer cells: a novel strategy to overcome drug resistance associated with mitochondrial respiratory defect and hypoxia. Cancer research. 2005;65(2):613-21.

7. Robey IF, Lien AD, Welsh SJ, Baggett BK, Gillies RJ. Hypoxia-inducible factor-1α and the glycolytic phenotype in tumors. Neoplasia. 2005;7(4):324-30.

8. Sakamoto T, Niiya D, Seiki M. Targeting the Warburg effect that arises in tumor cells expressing membrane type-1 matrix metalloproteinase. Journal of Biological Chemistry. 2011;286(16):14691-704.

9. Basseres D, Baldwin AS. Nuclear factor-κB and inhibitor of κB kinase pathways in oncogenic initiation and progression. Oncogene. 2006;25(51):6817-30.

10. Biswas DK, Iglehart JD. Linkage between EGFR family receptors and nuclear factor kappaB (NF‐κB) signaling in breast cancer. Journal of cellular physiology. 2006;209(3):645-52.

11. Ahmed KM, Cao N, Li JJ. HER-2 and NF-κB as the targets for therapy-resistant breast cancer. Anticancer research. 2006;26(6B):4235-43.

12. Bourguignon LY, Xia W, Wong G. Hyaluronan-mediated CD44 interaction with p300 and SIRT1 regulates β-catenin signaling and NFκB-specific transcription activity leading to MDR1 and Bcl-xL gene expression and chemoresistance in breast tumor cells. Journal of Biological Chemistry. 2009;284(5):2657-71.

13. Guttridge DC, Albanese C, Reuther JY, Pestell RG, Baldwin AS. NF-κB controls cell growth and differentiation through transcriptional regulation of cyclin D1. Molecular and cellular biology. 1999;19(8):5785-99.

14. Hinz M, Krappmann D, Eichten A, Heder A, Scheidereit C, Strauss M. NF-κB function in growth control: regulation of cyclin D1 expression and G0/G1-to-S-phase transition. Molecular and cellular biology. 1999;19(4):2690-8.

15. Toualbi-Abed K, Daniel F, Güller MC, Legrand A, Mauriz J-L, Mauviel A, et al. Jun D cooperates with p65 to activate the proximal κB site of the cyclin D1 promoter: role of PI3K/PDK-1. Carcinogenesis. 2008;29(3):536-43.

16. Moon D-O, Kim M-O, Kang S-H, Choi YH, Kim G-Y. Sulforaphane suppresses TNF-α-mediated activation of NF-κB and induces apoptosis through activation of reactive oxygen species-dependent caspase-3. Cancer letters. 2009;274(1):132-42.

17. Catz SD, Johnson JL. Transcriptional regulation of bcl-2 by nuclear factor κB and its significance in prostate cancer. Oncogene. 2001;20(50).

18. Wang F, Yang J-L, Yu K-k, Xu M, Xu Y-z, Chen L, et al. Activation of the NF-κB pathway as a mechanism of alcohol enhanced progression and metastasis of human hepatocellular carcinoma. Molecular cancer. 2015;14(1):1.

19. Dongare S, Gupta SK, Mathur R, Saxena R, Mathur S, Agarwal R, et al. Zingiber officinale attenuates retinal microvascular changes in diabetic rats via anti-inflammatory and antiangiogenic mechanisms. Molecular Vision. 2016;22:599.

20. Habib SHM, Makpol S, Hamid NAA, Das S, Ngah WZW, Yusof YAM. Ginger extract (Zingiber officinale) has anti-cancer and anti-inflammatory effects on ethionine-induced hepatoma rats. Clinics. 2008;63(6):807-13.

21. Roufogalis BD. Zingiber officinale (Ginger): a future outlook on its potential in prevention and treatment of diabetes and prediabetic states. New Journal of Science. 2014;2014.

22. Flores-López LA, Martínez-Hernández MG, Viedma-Rodríguez R, Díaz-Flores M, Baiza-Gutman LA. High glucose and insulin enhance uPA expression, ROS formation and invasiveness in breast cancer-derived cells. Cellular Oncology. 2016:1-14.

23. El Sayed SM, Mahmoud AA, El Sawy SA, Abdelaal EA, Fouad AM, Yousif RS, et al. Warburg effect increases steady-state ROS condition in cancer cells through decreasing their antioxidant capacities (anticancer effects of 3-bromopyruvate through antagonizing Warburg effect). Medical hypotheses. 2013;81(5):866-70.

24. Liberti MV, Locasale JW. The Warburg effect: how does it benefit cancer cells? Trends in biochemical sciences. 2016;41(3):211-8.

25. Végran F, Boidot R, Michiels C, Sonveaux P, Feron O. Lactate influx through the endothelial cell monocarboxylate transporter MCT1 supports an NF-κB/IL-8 pathway that drives tumor angiogenesis. Cancer research. 2011;71(7):2550-60.

26. Inoguchi T, Li P, Umeda F, Yu HY, Kakimoto M, Imamura M, et al. High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C--dependent activation of NAD (P) H oxidase in cultured vascular cells. Diabetes. 2000;49(11):1939-45.

27. Russel JW, Golovoy D, Vincent AM, Mahendru P, Olzmann JA, Mentzer A, et al. High glucose-induced oxidative stress and mitochondrial dysfunction in neurons. The FASEB Journal. 2002;16(13):1738-48.

28. Turturro F, Friday E, Welbourne T. Hyperglycemia regulates thioredoxin-ROS activity through induction of thioredoxin-interacting protein (TXNIP) in metastatic breast cancer-derived cells MDA-MB-231. BMC cancer. 2007;7(1):1.

29. Liou G-Y, Storz P. Reactive oxygen species in cancer. Free radical research. 2010;44(5):479-96.

30. Anrather J, Csizmadia V, Soares MP, Winkler H. Regulation of NF-κB RelA phosphorylation and transcriptional activity by p21 ras and protein kinase Cζ in primary endothelial cells. Journal of Biological Chemistry. 1999;274(19):13594-603.

31. Deng W-G, Zhu Y, Wu KK. Up-regulation of p300 binding and p50 acetylation in tumor necrosis factor-α-induced cyclooxygenase-2 promoter activation. Journal of Biological Chemistry. 2003;278(7):4770-7.

32. Hu J, Nakano H, Sakurai H, Colburn NH. Insufficient p65 phosphorylation at S536 specifically contributes to the lack of NF-κB activation and transformation in resistant JB6 cells. Carcinogenesis. 2004;25(10):1991-2003.

33. Sizemore N, Leung S, Stark GR. Activation of phosphatidylinositol 3-kinase in response to interleukin-1 leads to phosphorylation and activation of the NF-κB p65/RelA subunit. Molecular and cellular biology. 1999;19(7):4798-805.

34. Masur K, Vetter C, Hinz A, Tomas N, Henrich H, Niggemann B, et al. Diabetogenic glucose and insulin concentrations modulate transcriptom and protein levels involved in tumour cell migration, adhesion and proliferation. British journal of cancer. 2011;104(2):345-52.

35. Okumura M, Yamamoto M, Sakuma H, Kojima T, Maruyama T, Jamali M, et al. Leptin and high glucose stimulate cell proliferation in MCF-7 human breast cancer cells: reciprocal involvement of PKC-α and PPAR expression. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research. 2002;1592(2):107-16.

36. Zhu S, Yao F, Li W-H, Wan J-N, Zhang Y-M, Tang Z, et al. PKCδ-dependent activation of the ubiquitin proteasome system is responsible for high glucose-induced human breast cancer MCF-7 cell proliferation, migration and invasion. Asian Pacific Journal of Cancer Prevention. 2013;14(10):5687-92.

How to Cite
Nasir Kansestani A, Mansouri K, Hemmati S, Zare ME, Moatafaei A. High Glucose-reduced Apoptosis in Human Breast Cancer Cells Is Mediated by Activation of NF-κB. Iran J Allergy Asthma Immunol. 18(2):153-162.
Original Article(s)