Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Tumor Biology
Published in: Tumor Biology 1/2013

01-02-2013 | Research Article

Part I. Molecular and cellular characterization of high nitric oxide-adapted human breast adenocarcinoma cell lines

Authors: B. J. Vesper, A. Onul, G. K. Haines III, G. Tarjan, J. Xue, K. M. Elseth, B. Aydogan, M. B. Altman, J. C. Roeske, W. A. Paradise, H. De Vitto, J. A. Radosevich

Published in: Tumor Biology | Issue 1/2013

Login to get access

Abstract

There is a lack of understanding of the casual mechanisms behind the observation that some breast adenocarcinomas have identical morphology and comparatively different cellular growth behavior. This is exemplified by a differential response to radiation, chemotherapy, and other biological intervention therapies. Elevated concentrations of the free radical nitric oxide (NO), coupled with the up-regulated enzyme nitric oxide synthase (NOS) which produces NO, are activities which impact tumor growth. Previously, we adapted four human breast cancer cell lines: BT-20, Hs578T, T-47D, and MCF-7 to elevated concentrations of nitric oxide (or high NO [HNO]). This was accomplished by exposing the cell lines to increasing levels of an NO donor over time. Significantly, the HNO cell lines grew faster than did each respective (“PARENT”) cell line even in the absence of NO donor-supplemented media. This was evident despite each “parent” being morphologically equivalent to the HNO adapted cell line. Herein, we characterize the HNO cells and their biological attributes against those of the parent cells. Pairs of HNO/parent cell lines were then analyzed using a number of key cellular activity criteria including: cell cycle distribution, DNA ploidy, response to DNA damage, UV radiation response, X-ray radiation response, and the expression of significant cellular enzymes. Other key enzyme activities studied were NOS, p53, and glutathione S-transferase-pi (GST-pi) expression. HNO cells were typified by a far more aggressive pattern of growth and resistance to various treatments than the corresponding parent cells. This was evidenced by a higher S-phase percentage, variable radioresistance, and up-regulated GST-pi and p53. Taken collectively, this data provides evidence that cancer cells subjected to HNO concentrations become resistant to free radicals such as NO via up-regulated cellular defense mechanisms, including p53 and GST-pi. The adaptation to NO may explain how tumor cells acquire a more aggressive tumor phenotype.
Literature
1.
2.
Jemal A, et al. Global cancer statistics. CA: A Cancer Journal for Clinicians. 2011;61(2):69–90.CrossRef
3.
Bentz BG, et al. Nitric oxide synthase type 3 is increased in squamous hyperplasia, dysplasia, and squamous cell carcinoma of the head and neck. Ann Otol Rhinol Laryngol. 1999;108(8):781–7.
4.
5.
Singh S, et al. The impact of intracellular generation of nitric oxide on the radiation response of human tumor cells. Radiat Res. 2009;171(5):572–80.PubMedCrossRef
6.
Paradise WA, Vesper BJ, Goel A, Waltonen JD, Altman KW, Haines III GK, Radosevich JA. Nitric oxide: perspectives and emerging studies of a well known cytotoxin. Int J Mol Sci. 2010;11:2715–45.PubMedCrossRef
7.
Vesper BJ, et al. Long-term adaptation of breast tumor cell lines to high concentrations of nitric oxide. Tumor Biol. 2010;31(4):267–75.CrossRef
8.
Bentz BG, et al. Nitrosative stress induces DNA strand breaks but not caspase mediated apoptosis in a lung cancer cell line. J Carcinog. 2004;3(1):16.PubMedCrossRef
9.
Bentz BG, et al. Cytoplasmic localization of endothelial constitutive nitric oxide synthase in endometrial carcinomas. Tumor Biol. 1997;18(5):290–300.CrossRef
10.
Bentz BG, et al. Endothelial constitutive nitric oxide synthase (ecNOS) localization in normal and neoplastic salivary tissue. Head Neck. 1998;20(4):304–9.PubMedCrossRef
11.
Bentz BG, et al. The kinetics and redox state of nitric oxide determine the biological consequences in lung adenocarcinoma. Tumor Biol. 2007;28(6):301–11.CrossRef
12.
Bentz BG, et al. The yin and yang of nitric oxide: reflections on the physiology and pathophysiology of NO. Head Neck. 2000;22(1):71–83.PubMedCrossRef
13.
Keydar I, et al. Establishment and characterization of a cell line of human breast carcinoma origin. Eur J Cancer. 1979;15(5):659–70.PubMed
14.
Beevi SS, Rasheed MH, Geetha A. Evidence of oxidative and nitrosative stress in patients with cervical squamous cell carcinoma. Clin Chim Acta. 2007;375(1–2):119–23.PubMedCrossRef
15.
Loibl S, et al. Expression of endothelial and inducible nitric oxide synthase in benign and malignant lesions of the breast and measurement of nitric oxide using electron paramagnetic resonance spectroscopy. Cancer. 2002;95(6):1191–8.PubMedCrossRef
16.
Duenas-Gonzalez A, et al. Expression of inducible nitric oxide synthase in breast cancer correlates with metastatic disease. Mod Pathol. 1997;10(7):645–9.PubMed
17.
Tschugguel W, et al. Expression of inducible nitric oxide synthase in human breast cancer depends on tumor grade. Breast Cancer Res Treat. 1999;56(2):145–51.PubMedCrossRef
18.
Cobbs CS, et al. Expression of nitric oxide synthase in human central nervous system tumors. Cancer Res. 1994;55(4):727–30.
19.
Chandra R, et al. Expression of nitric oxide synthase type 3 in reflux-induced esophageal lesions. Otolaryngol. Head Neck Surg. 2001;124(4):442–7.CrossRef
20.
Ambs S, et al. Frequent nitric oxide synthase-2 expression in human colon adenomas: implication for tumor angiogenesis and colon cancer progression. Cancer Res. 1998;58(2):334–41.PubMed
21.
Chen YK, Hsue SS, Lin LM. Increased expression of inducible nitric oxide synthase for human buccal squamous-cell carcinomas: immunohistochemical, reverse transcription-polymerase chain reaction (RT-PCR) and in situ RT-PCR studies. Head Neck. 2002;24(10):925–32.PubMedCrossRef
22.
Hibbs JB, et al. Nitric oxide: a cytotoxic activated macrophage effector molecule. Biochem Biophys Res Commun. 1988;157(1):87–94.PubMedCrossRef
23.
Hibbs JB, Vavrin A, Taintor RR. l-Arginine is required for expression of the activated macrophage effector mechanism causing selective metabolic inhibition in target cells. J Immunol. 1987;138(2):550–65.
24.
Loibl S, et al. The role of early expression of inducible nitric oxide synthase in human breast cancer. Eur J Cancer. 2005;41(2):265–71.PubMedCrossRef
25.
26.
Altman MB, et al. Characterization of a novel phantom for three-dimensional in vitro cell experiments. Phys Med Biol. 2009;54(5):N75–82.PubMedCrossRef
27.
Natarajan N, et al. Adaptation of the diphenylamine (DPA) assay to a 96-well plate tissue culture format and comparison with the MTT assay. BioTechniques. 1994;17(1):166–71.PubMed
28.
Smyth, G.K., et al., Limma: Linear Models for Microarray Data. Bioinformatics and computationalbBiology solutions using R and bioconductor, 2005: 397–420.
29.
Ritchie ME, et al. A comparison of background correction methods for two-colour microarrays. Bioinformatics. 2007;23(20):2700–7.PubMedCrossRef
30.
31.
Smyth, G.K., Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol, 2004. 3: Article3.
32.
Blackett NM, et al. Radiation modifying effect of the free radical norpseudopelletierene-N-oxyl on normal bone marrow stem cells in vitro and in vivo. Radiat Res. 1974;58(3):361–72.PubMedCrossRef
33.
Bristow, R.G. and R.P. Hill, Molecular and cellular basis of radiotherapy. In I.F. Tannock and R.P. Hill, editors. The basic science of oncology. 1998, McGraw-Hill: New York. p. 95–321.
34.
Chen HHW, et al. Increased expression of nitric oxide synthase and cyclooxygenase-2 is associated with poor survival in cervical cancer treated with radiotherapy. Int J Radiation Oncology Biol Phys. 2005;63(4):1093–100.
35.
Cooke BC, Fielden EM, Johnson M. Polyfunctional radiosensitizers: I. Effects of a nitroxyl biradical on the survival of mammalian cells in vitro. Radiat Res. 1976;65(1):152–62.PubMedCrossRef
36.
De Ridder M, et al. Hypoxic tumor cell radiosensitization through nitric oxide. Nitric Oxide. 2008;19(2):164–9.PubMedCrossRef
37.
George R, et al. Audio-visual biofeedback for respiratory-gated radiotherapy: impact of audio instruction and audio-visual biofeedback on respiratory-gated radiotherapy. Int J Radiat Oncol Biol Phys. 2006;65(3):924–33.PubMedCrossRef
38.
Griffin RJ, et al. Radiosensitization of hypoxic tumor cells in vitro by nitric oxide. Int J Radiat Oncol Biol Phys. 1996;36(2):377–83.PubMedCrossRef
39.
Hogle WP. Radiation therapy in the treatment of breast cancer. Semin Oncol Nurs. 2007;23(1):20–8.
40.
41.
Wang YZ, et al. Expression of nitric oxide synthase in human gastric carcinoma and its relation to p53, PCNA. World J Gastroenterol. 2005;11(1):46–50.PubMed
42.
Glynn, S., et al., iNOS expression is associated with basal-like breast cancer phenotype and predicts poor survival in ERneg breast cancer. Nitric Oxide, 2007. 17(Supp. 1): S17.
43.
Nakamura Y, et al. Nitric oxide in breast cancer: induction of vascular endothelial growth factor-C and correlation with metastasis and poor prognosis. Clin Cancer Res. 2006;12(4):1201–7.PubMedCrossRef
44.
Thomsen LL, et al. Nitric oxide synthase in human breast cancer. Br J Cancer. 1995;72(1):41–4.
45.
Bulut AS, et al. Significance of inducible nitric oxide synthase expression in benign and malignant breast epithelium: an immunohistochemical study of 151 cases. Virchows Arch. 2005;447(1):24–30.PubMedCrossRef
46.
Masri FA, et al. Abnormalities in nitric oxide and its derivatives in lung cancer. Am J Respir Crit Care Med. 2005;172(5):597–605.PubMedCrossRef
47.
Lee TW, et al. Differential expression of inducible nitric oxide synthase and peroxisome proliferator-activated receptor gamma in non-small cell lung carcinoma. Eur J Cancer. 2003;39(9):1296–301.PubMedCrossRef
48.
Liu CY, et al. Increased level of exhaled nitric oxide and up-regulation of inducible nitric oxide synthase in patients with primary lung cancer. Br J Cancer. 1998;78(4):534–41.PubMedCrossRef
49.
Rosbe KW, et al. Immunohistochemical characterization of nitric oxide synthase activity in squamous cell carcinoma of the head and neck. otolaryngol. Head Neck Surg. 1995;113(5):541–9.
50.
Yarmolyuk Y, et al. A model system for dtudying nitric oxide in tumors: high nitric oxide-adapted head and neck squamous cell lines. Tumor Biol. 2011;32(1):77–85.CrossRef
51.
Bentz BG, et al. Nitric oxide and apoptosis during human head and neck squamous cell carcinoma development. Am J Otolaryngol. 2002;23(1):4–11.PubMedCrossRef
52.
Gavilanes J, et al. Nitric oxide synthase activity in human squamous cell carcinoma of the head and neck. Laryngoscope. 1999;109(1):148–52.PubMedCrossRef
53.
Bentz BG, Haines III GK, Radosevich JA. Glutathione S-transferase pi in squamous cell carcinoma of the head and neck. Laryngoscope. 2000;110(10 Pt 1):1642–7.PubMedCrossRef
54.
Cullen KJ, et al. Glutathione S-transferase pi amplification is associated with cisplatin resistance in head and neck squamous cell carcinoma cell lines and primary tumors. Cancer Res. 2003;63(23):8097–102.PubMed
55.
Shiga H, et al. Prognostic value of p53, glutathione S-transferase pi, and thymidylate synthase for neoadjuvant cisplatin-based chemotherapy in head and neck cancer. Clin Cancer Res. 1999;5(12):4097–104.PubMed
56.
Dourado DF, Fernandes A, Ramos MJ. Mammalian cytosolic glutathione transferases. Curr Protein Pept Sci. 2008;9(4):325–37.PubMedCrossRef
57.
Mulder TP, et al. Glutatione S-transferases and glutathione in human head and neck cancer. Carcinogenesis. 1995;16(3):619–24.PubMedCrossRef
58.
De Vitto, et al. Part II. Mitochondrial mutational status of high nitric oxide adapted cell line BT-20 (BT-20-HNO) as it relates to human primary breast tumors. Tumor Biol. 2012. doi:10.​1007/​s13277-012-0555-4.
59.
De Vitto, et al. Part III. Molecular changes induced by high nitric oxide adaptation in human breast cancer cell line BT-20 (BT-20-HNO): a switch from aerobic to anaerobic. Tumor Biol. 2012. doi:10.​1007/​s13277-012-0564-3.
Metadata
Title
Part I. Molecular and cellular characterization of high nitric oxide-adapted human breast adenocarcinoma cell lines
Authors
B. J. Vesper
A. Onul
G. K. Haines III
G. Tarjan
J. Xue
K. M. Elseth
B. Aydogan
M. B. Altman
J. C. Roeske
W. A. Paradise
H. De Vitto
J. A. Radosevich
Publication date
01-02-2013
Publisher
Springer Netherlands
Published in
Tumor Biology / Issue 1/2013
Print ISSN: 1010-4283
Electronic ISSN: 1423-0380
DOI
https://doi.org/10.1007/s13277-012-0530-0

Other articles of this Issue 1/2013

Tumor Biology 1/2013 Go to the issue

Research Article

Circulating immunogenic cell death biomarkers HMGB1 and RAGE in breast cancer patients during neoadjuvant chemotherapy

Research Article

Associations between high levels of Notch1 expression and high invasion and poor overall survival in hepatocellular carcinoma

Research Article

Heterozygosity for TGF β1 -509C/T Polymorphism is associated with risk for breast cancer in South Indian population

Research Article

Methylation of the p73 gene in patients with myelodysplastic syndromes: correlations with apoptosis and prognosis

Research Article

Part II. Mitochondrial mutational status of high nitric oxide adapted cell line BT-20 (BT-20-HNO) as it relates to human primary breast tumors

Research Article

Development and validation of H11B2C2 monoclonal antibody-reactive hyaluronic acid binding protein: overexpression of HABP during human tumor progression
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits