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Upregulation of NOX-2 and Nrf-2 Promotes 5-Fluorouracil Resistance of Human Colon Carcinoma (HCT-116) Cells

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Abstract

Altered expression of cellular redox genes and proteins contributes to invasion, metastasis, and drug resistance in cancer. NADPH oxidase (NOX) isoforms are the pro-oxidant enzymes that generate ROS as a primary product. Dysregulation of NOX activity and expression alters ROS generation, which either directly or indirectly modulates cell death and survival signaling during the progression of cancer. Nuclear factor erythroid 2-related factor 2 (Nrf-2) is an inducible transcription factor, which transcribes an array of antioxidant genes and protects cancer cells from the oxidative stress. Both NOXs and Nrf-2 participate in the regulation of cellular redox homeostasis; but their dysregulation promotes oxidative stress, which contributes to the progression of different types of cancer. Indeed, the role of NOX isoforms and Nrf-2 in developing the drug resistance in cancer is largely unknown. In the present study, we have explored the association of NOX isoforms and Nrf-2 signaling with the MDR1 gene expression in colon carcinoma cells (HCT-116/R). The MDR1 gene was overexpressed to develop resistant HCT-116/R cells and the NOX activation and ROS generation were monitored. We also assessed the role of NOX isoforms and Nrf-2 in the 5-fluorouracil (5-FU) mediated apoptotic cell death of HCT-116/R cells. The HCT-116/R cells demonstrated higher expression of HIF-1α, Nrf-2, and HO-1 and were highly resistant to 5-FU; they also displayed upregulated expression and activity of NOX-2, as well as elevated ROS levels. Interestingly, the treatment with HDC, a specific NOX-2 inhibitor, reduced the ROS levels in HCT-116/R cells. The treatment with HDC and ML-385 (specific inhibitor of Nrf-2) augmented the 5-FU-mediated apoptotic cell death of HCT-116/R cells, which suggests that NOX-2 and Nrf-2 are involved in the development of the chemoresistant phenotype of these cells. Taken together, NOX-2 and Nrf-2 are associated with developing drug resistance of colorectal cancer cells and might be potential targets to overcome drug resistance during cancer therapy.

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Abbreviations

5-FU:

5-fluorouracil

ABC:

ATP-binding cassette

DPBS:

Dulbecco’s phosphate buffered saline

HCT-116/R:

MDR resistant colon cancer cells

NOX:

NADPH oxidase

Nrf-2:

nuclear factor erythroid 2-related factor 2

ROS:

reactive oxygen species

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Acknowledgements

The authors are grateful to the Indian Institute of Advanced Research, for providing the infrastructure and the research facility.

Funding

This study was supported by the SERB, Department of Science & Technology, New Delhi, Government of India (research grant EMR/2016/002574 to C. P.); ICMR, Gov of India (Senior Research fellowship to F. U. V.) and DST-INSPIRE (fellowship to B. N. W.).

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Authors and Affiliations

  1. School of Biological Sciences & Biotechnology, Indian Institute of Advanced Research, Koba Institutional Area, 382426, Gandhinagar, Gujarat, India

    Bhargav N. Waghela, Foram U. Vaidya & Chandramani Pathak

  2. Amity Institute of Biotechnology, Amity University Haryana, 122413, Gurgaon, India

    Chandramani Pathak

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  1. Bhargav N. Waghela
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  2. Foram U. Vaidya
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  3. Chandramani Pathak
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Correspondence to Chandramani Pathak.

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The authors declare no conflict of interest. This article does not contain description of studies with the involvement of humans or animal subjects.

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Waghela, B.N., Vaidya, F.U. & Pathak, C. Upregulation of NOX-2 and Nrf-2 Promotes 5-Fluorouracil Resistance of Human Colon Carcinoma (HCT-116) Cells. Biochemistry Moscow 86, 262–274 (2021). https://doi.org/10.1134/S0006297921030044

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