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1. Nutritional Stress in Head and Neck Cancer Originating Cell Lines: The Sensitivity of the NRF2-NQO1 Axis

Author

Lidija Milković, Marko Tomljanović, Ana Čipak Gašparović, Renata Novak Kujundžić, Dina Šimunić, Paško Konjevoda, Anamarija Mojzeš, Nikola Đaković, Neven Žarković, and Koraljka Gall Trošelj

Subjects
glucose deprivation, glutamine deprivation, viability, proliferation, ROS, NRF2-NQO1 axis, IMR-90, NQO1 transcript variants, rs1800566, TP53 mutation, Cytology, QH573-671
Abstract

Nutritional stress disturbs the cellular redox-status, which is characterized by the increased generation of reactive oxygen species (ROS). The NRF2-NQO1 axis represents a protective mechanism against ROS. Its strength is cell type-specific. FaDu, Cal 27 and Detroit 562 cells differ with respect to basal NQO1 activity. These cells were grown for 48 hours in nutritional conditions (NC): (a) Low glucose−NC2, (b) no glucose, no glutamine−NC3, (c) no glucose with glutamine−NC4. After determining the viability, proliferation and ROS generation, NC2 and NC3 were chosen for further exploration. These conditions were also applied to IMR-90 fibroblasts. The transcripts/transcript variants of NRF2 and NQO1 were quantified and transcript variants were characterized. The proteins (NRF2, NQO1 and TP53) were analyzed by a western blot in both cellular fractions. Under NC2, the NRF2-NQO1 axis did not appear activated in the cancer cell lines. Under NC3, the NRF2-NQO1axis appeared slightly activated in Detroit 562. There are opposite trends with respect to TP53 nuclear signal when comparing Cal 27 and Detroit 562 to FaDu, under NC2 and NC3. The strong activation of the NRF2-NQO1 axis in IMR-90 resulted in an increased expression of catalytically deficient NQO1, due to NQO1*2/*2 polymorphism (rs1800566). The presented results call for a comprehensive exploration of the stress response in complex biological systems.

Published
2019
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2. Nutritional Stress in Head and Neck Cancer Originating Cell Lines: The Sensitivity of the NRF2-NQO1 Axis

Author

Neven Žarković, Ana Čipak Gašparović, Dina Šimunić, Paško Konjevoda, Nikola Đaković, Renata Novak Kujundžić, Marko Tomljanović, Lidija Milkovic, Koraljka Gall Trošelj, and Anamarija Mojzeš

Subjects
Glutamine, Cell, Cell Culture Techniques, rs1800566, medicine.disease_cause, Fight-or-flight response, 0302 clinical medicine, NAD(P)H Dehydrogenase (Quinone), glutamine deprivation, lcsh:QH301-705.5, chemistry.chemical_classification, 0303 health sciences, medicine.diagnostic_test, Chemistry, glucose deprivation, TP53 mutation, Head and Neck Neoplasms / pathology, ROS, General Medicine, 3. Good health, medicine.anatomical_structure, Head and Neck Neoplasms, 030220 oncology & carcinogenesis, NF-E2-Related Factor 2 / metabolism, Cancer cell lines, Head and Neck Neoplasms / metabolism, Cell Culture Techniques / methods, NF-E2-Related Factor 2, proliferation, Article, Cell Line, 03 medical and health sciences, Western blot, IMR-90, Glutamine / deficiency, medicine, Humans, NAD(P)H Dehydrogenase (Quinone) / metabolism, 030304 developmental biology, Reactive oxygen species, viability, Head and neck cancer, NRF2-NQO1 axis, NQO1 transcript variants, Reactive Oxygen Species / metabolism, Basic Medical Sciences, Glucose / deficiency, medicine.disease, Molecular biology, Oxidative Stress, Glucose, lcsh:Biology (General), Cell culture, Reactive Oxygen Species, Oxidative stress
Abstract

Nutritional stress disturbs the cellular redox-status, which is characterized by the increased generation of reactive oxygen species (ROS). The NRF2-NQO1 axis represents a protective mechanism against ROS. Its strength is cell type-specific. FaDu, Cal 27 and Detroit 562 cells differ with respect to basal NQO1 activity. These cells were grown for 48 hours in nutritional conditions (NC): (a) Low glucose&ndash, NC2, (b) no glucose, no glutamine&ndash, NC3, (c) no glucose with glutamine&ndash, NC4. After determining the viability, proliferation and ROS generation, NC2 and NC3 were chosen for further exploration. These conditions were also applied to IMR-90 fibroblasts. The transcripts/transcript variants of NRF2 and NQO1 were quantified and transcript variants were characterized. The proteins (NRF2, NQO1 and TP53) were analyzed by a western blot in both cellular fractions. Under NC2, the NRF2-NQO1 axis did not appear activated in the cancer cell lines. Under NC3, the NRF2-NQO1axis appeared slightly activated in Detroit 562. There are opposite trends with respect to TP53 nuclear signal when comparing Cal 27 and Detroit 562 to FaDu, under NC2 and NC3. The strong activation of the NRF2-NQO1 axis in IMR-90 resulted in an increased expression of catalytically deficient NQO1, due to NQO1*2/*2 polymorphism (rs1800566). The presented results call for a comprehensive exploration of the stress response in complex biological systems.

Published
2019
Full Text
View/download PDF

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