Drug Res (Stuttg) 2017; 67(01): 13-19
DOI: 10.1055/s-0042-115405
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Co-inhibition of Notch and NF-κB Signaling Pathway Decreases Proliferation through Downregulating IκB-α and Hes-1 Expression in Human Ovarian Cancer OVCAR-3 Cells

M. Majidinia
1   Department of Clinical Biochemistry and Laboratory Medicine, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
2   Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
E. Alizadeh
3   Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran
B. Yousefi
1   Department of Clinical Biochemistry and Laboratory Medicine, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
M. Akbarzadeh
4   Department of Biochemistry, Higher Education Institute of Rab-Rashid, Tabriz, Iran
A. Mihanfar
2   Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
M. Rahmati-Yamchi
1   Department of Clinical Biochemistry and Laboratory Medicine, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
N. Zarghami
1   Department of Clinical Biochemistry and Laboratory Medicine, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran
5   Hematology and Oncology Research Center, Tabriz University of Medical Science, Tabriz, Iran
› Author Affiliations
Further Information

Publication History

received 15 May 2016

accepted 25 July 2016

Publication Date:
29 September 2016 (online)


Background: Ovarian cancer is one of the most lethal gynecological malignancies and numerous changes in signaling cascades are involved in the initiation and progression of ovarian cancerous cells. Here, we investigated the role of NF-κB and Notch pathways inhibition on human ovarian cancer OVCAR-3 cells proliferation and IκB-α and Hes-1 expression as 2 key genes in these pathways regulation.

Methods: The effects of Bay 11-7085 and DAPT, NF-κB and Notch pathways specific inhibitors, on cell proliferation were evaluated using MTT assay. In addition, the cells were transfected by Notch and IKK-β siRNAs. mRNA and protein levels of target genes were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot after 48 h incubation with inhibitors and siRNAs.

Results: Bay 11-7085 and DAPT significantly decreased the cell proliferation OVCAR-3. IκB-α and Hes-1 mRNA levels decreased to 5 or 3% and 6% or 2% after treatment with Bay 11-7085 or DAPT, respectively (p<0.05). We also found that combination treatment exert a more potent effects on the expression of these gene (p<0.05). Moreover, siRNA transfection caused a significant reduction in IκB-α and Hes-1 mRNA levels (p<0.05). In the protein level, OVCAR-3 cell treatment with both chemichal inhibitors and specific siRNA cause a significant decrease in the expression of target genes (p<0.05)

Conclusion: Our findings suggest that inhibition of NF-κB and Notch signaling pathways can effectively reduce OVCAR-3 cells proliferation. Therefore, pharmacological targeting of the NF-κB and Notch signaling pathway could be a promising future treatment of ovarian cancer.

  • References

  • 1 Kurman RJ, Shih I-M. The Origin and pathogenesis of epithelial ovarian cancer – a proposed unifying theory. The American journal of surgical pathology 2010; 34: 433
  • 2 Nezhat FR, Apostol R, Nezhat C et al. New insights in the pathophysiology of ovarian cancer and implications for screening and prevention. American journal of obstetrics and gynecology 2015; 213: 262-267
  • 3 Block MS, Maurer MJ, Goergen K et al. Plasma immune analytes in patients with epithelial ovarian cancer. Cytokine 2015; 73: 108-113
  • 4 Yousefi B, Rahmati M, Ahmadi Y. The roles of p53R2 in cancer progression based on the new function of mutant p53 and cytoplasmic p21. Life Sciences 2014; 99: 14-17
  • 5 Hoesel B, Schmid JA. The complexity of NF-κB signaling in inflammation and cancer. Molecular cancer 2013; 12: 1
  • 6 Bradford JW, Baldwin AS. IKK/nuclear factor-kappaB and oncogenesis: roles in tumor-initiating cells and in the tumor microenvironment. Adv Cancer Res 2014; 121: 125-145
  • 7 Dolcet X, Llobet D, Pallares J et al. NF-kB in development and progression of human cancer. Virchows archiv 2005; 446: 475-482
  • 8 Karimian A, Ahmadi Y, Yousefi B. Multiple functions of p21 in cell cycle, apoptosis and transcriptional regulation after DNA damage. DNA Repair (Amst) 2016; 42: 63-71
  • 9 Hernandez L, Hsu SC, Davidson B et al. Activation of NF-κB signaling by inhibitor of NF-κB kinase β increases aggressiveness of ovarian cancer. Cancer research 2010; 70: 4005-4014
  • 10 Yen W-C, Fischer MM, Axelrod F et al. Targeting Notch signaling with a Notch2/Notch3 antagonist (tarextumab) inhibits tumor growth and decreases tumor-initiating cell frequency. Clinical Cancer Research 2015; 21: 2084-2095
  • 11 Takebe N, Miele L, Harris PJ et al. Targeting Notch, Hedgehog, and Wnt pathways in cancer stem cells: clinical update. Nature reviews Clinical oncology 2015; 12: 445-464
  • 12 Previs RA, Coleman RL, Harris AL et al. Molecular pathways: translational and therapeutic implications of the Notch signaling pathway in cancer. Clinical Cancer Research 2015; 21: 955-961
  • 13 Kageyama R, Shimojo H, Imayoshi I. Dynamic expression and roles of Hes factors in neural development. Cell and tissue research 2015; 359: 125-133
  • 14 Choi J-H, Park JT, Davidson B et al. Jagged-1 and Notch3 juxtacrine loop regulates ovarian tumor growth and adhesion. Cancer research 2008; 68: 5716-5723
  • 15 Park JT, Li M, Nakayama K et al. Notch3 gene amplification in ovarian cancer. Cancer research 2006; 66: 6312-6318
  • 16 Kumar V, Palermo R, Talora C et al. Notch and NF-kB signaling pathways regulate miR-223/FBXW7 axis in T-cell acute lymphoblastic leukemia. Leukemia 2014; 28: 2324-2335
  • 17 Jiao X, Wood LD, Lindman M et al. Somatic mutations in the notch, NF-KB, PIK3CA, and hedgehog pathways in human breast cancers. Genes, Chromosomes and Cancer 2012; 51: 480-489
  • 18 Coward JI, Middleton K, Murphy F. New perspectives on targeted therapy in ovarian cancer. International journal of women’s health 2015; 7: 189
  • 19 Annunziata CM, Stavnes HT, Kleinberg L et al. Nuclear factor κB transcription factors are coexpressed and convey a poor outcome in ovarian cancer. Cancer 2010; 116: 3276-3284
  • 20 Sagher E, Hernandez L, Heywood C et al. The small molecule NSC676914A is cytotoxic and differentially affects NFκB signaling in ovarian cancer cells and HEK293 cells. Cancer cell international 2014; 14: 1
  • 21 McAuliffe SM, Morgan SL, Wyant GA et al. Targeting Notch, a key pathway for ovarian cancer stem cells, sensitizes tumors to platinum therapy. Proceedings of the National Academy of Sciences 2012; 109: E2939-E2948
  • 22 Kotsopoulos IC, Papanikolaou A, Lambropoulos AF et al. Serous ovarian cancer signaling pathways. International Journal of Gynecological Cancer 2014; 24: 410-417
  • 23 Groeneweg JW, Hall TR, Zhang L et al. Inhibition of gamma-secretase activity impedes uterine serous carcinoma growth in a human xenograft model. Gynecologic oncology 2014; 133: 607-615
  • 24 Chiaramonte R, Colombo M, Bulfamante G et al. Notch pathway promotes ovarian cancer growth and migration via CXCR4/SDF1α chemokine system. The international journal of biochemistry & cell biology 2015; 66: 134-140
  • 25 Groeneweg JW, Foster R, Growdon WB et al. Notch signaling in serous ovarian cancer. Journal of ovarian research 2014; 7: 1
  • 26 Espinosa L, Cathelin S, D’Altri T et al. The Notch/Hes1 pathway sustains NF-κB activation through CYLD repression in T-cell leukemia. Cancer cell 2010; 18: 268-281
  • 27 Maniati E, Bossard M, Cook N et al. Crosstalk between the canonical NF-κB and Notch signaling pathways inhibits Pparγ expression and promotes pancreatic cancer progression in mice. The Journal of clinical investigation 2011; 121: 4685-4699