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Optical Redox Imaging Detects the Effects of DEK Oncogene Knockdown on the Redox State of MDA-MB-231 Breast Cancer Cells

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Abstract

Purpose

Optical redox imaging (ORI), based on collecting the endogenous fluorescence of reduced nicotinamide adenine dinucleotide (NADH) and oxidized flavoproteins (Fp) containing a redox cofactor flavin adenine dinucleotide (FAD), provides sensitive indicators of cellular metabolism and redox status. ORI indices (such as NADH, FAD, and their ratio) have been under investigation as potential progression/prognosis biomarkers for cancer. Higher FAD redox ratio (i.e., FAD/(FAD + NADH)) has been associated with higher invasive/metastatic potential in tumor xenografts and cultured cells. This study is to examine whether ORI indices can respond to the modulation of oncogene DEK activities that change cancer cell invasive/metastatic potential.

Procedures

Using lentiviral shRNA, DEK gene expression was efficiently knocked down in MDA-MB-231 breast cancer cells (DEKsh). These DEKsh cells, along with scrambled shRNA-transduced control cells (NTsh), were imaged with a fluorescence microscope. In vitro invasive potential of the DEKsh cells and NTsh cells was also measured in parallel using the transwell assay.

Results

FAD and FAD redox ratios in polyclonal cells with DEKsh were significantly lower than that in NTsh control cells. Consistently, the DEKsh cells demonstrated decreased invasive potential than their non-knockdown counterparts NTsh cells.

Conclusions

This study provides direct evidence that oncogene activities could mediate ORI-detected cellular redox state.

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Acknowledgments

The authors thank Ms. Jinxia Jiang for her assistance in part of the data acquisition and Dr. Zhenwu Lin for valuable discussion and support. The authors would also like to appreciate the valuable discussion with and support from Dr. Andrea Stout and Ms. Jasmine Zhao of Cell and Developmental Biology Microscopy Core, Perelman School of Medicine, University of Pennsylvania.

Funding

This work was financially supported by the NIH Grant R01CA191207 (L.Z. Li) and NIH grant R37CA218072 (LMPV).

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Authors and Affiliations

  1. Rutgers Cancer Institute of New Jersey, New Brunswick, NJ, 08903, USA

    Yu Wen

  2. Department of Radiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Yu Wen, He N. Xu, Min Feng & Lin Z. Li

  3. Cincinnati Children’s Hospital Medical Center, Cancer and Blood Diseases Institute, Cincinnati, OH, USA

    Lisa Privette Vinnedge

  4. Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, USA

    Lisa Privette Vinnedge

  5. Abramson Cancer Center and Institute of Translational Medicine and Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Lin Z. Li

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  1. Yu Wen
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  2. He N. Xu
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  3. Lisa Privette Vinnedge
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  4. Min Feng
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Correspondence to Lin Z. Li.

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Wen, Y., Xu, H.N., Privette Vinnedge, L. et al. Optical Redox Imaging Detects the Effects of DEK Oncogene Knockdown on the Redox State of MDA-MB-231 Breast Cancer Cells. Mol Imaging Biol 21, 410–416 (2019). https://doi.org/10.1007/s11307-019-01321-w

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  • DOI: https://doi.org/10.1007/s11307-019-01321-w

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