Subcellular Analysis of Nuclear and Cytoplasmic Redox Indices Differentiates Breast Cancer Cell Subtypes Better Than Nuclear-to-cytoplasmic Area Ratio

Overview

Journal J Biomed Opt

Specialties Biomedical Engineering
Ophthalmology

Date 2022 Aug 10

PMID 35945669

Authors

Annemarie Jacob

He N Xu

Andrea L Stout

Lin Z Li

Affiliations

Soon will be listed here.

Abstract

Significance: Stratification of malignancy is valuable for cancer treatment. Both optical redox imaging (ORI) indices and nuclear-to-cytoplasmic volume/area ratio (N:C ratio) have been investigated to differentiate between cancers with varying aggressiveness, but these two methods have not been directly compared. The redox status in the cell nucleus has not been studied by ORI, and it remains unknown whether nuclear ORI indices add new biological information.

Aim: We sought to compare the capacity of whole-cell and subcellular ORI indices and N:C ratio to differentiate between breast cancer subtypes with varying aggressiveness and between mitotic and nonmitotic cells.

Approach: ORI indices for whole cell, cytoplasm, and nucleus as well as the N:C area ratio were generated for two triple-negative (more aggressive) and two receptor-positive (less aggressive) breast cancer cell lines by fluorescence microscopy.

Results: We found positive correlations between nuclear and cytoplasmic ORI indices within individual cells. On average, a nuclear redox status was found to be more oxidized than cytoplasm in triple-negative cells but not in receptor-positive cells. Whole-cell and subcellular ORI indices distinguished between the receptor statuses better than the N:C ratio. However, N:C ratio was a better differentiator between nonmitotic and mitotic triple-negative cells.

Conclusions: Subcellular ORI analysis differentiates breast cancer subtypes with varying aggressiveness better than N:C area ratio.

Citing Articles

Differential Mitochondrial Redox Responses to the Inhibition of NAD Salvage Pathway of Triple Negative Breast Cancer Cells.

Kollmar J, Xu J, Gonzalves D, Baur J, Li L, Tchou J Cancers (Basel). 2025; 17(1.

PMID: 39796638 PMC: 11718843. DOI: 10.3390/cancers17010007.

Multimodal Imaging Unveils the Impact of Nanotopography on Cellular Metabolic Activities.

Li Z, Sarikhani E, Prayotamornkul S, Meganathan D, Jahed Z, Shi L Chem Biomed Imaging. 2024; 2(12):825-834.

PMID: 39735831 PMC: 11672213. DOI: 10.1021/cbmi.4c00051.

Passage dependence of NADH redox status and reactive oxygen species level in triple-negative breast cancer cell lines with different invasiveness.

Podsednik A, Xu H, Li L Transl Breast Cancer Res. 2024; 5:27.

PMID: 39534579 PMC: 11557164. DOI: 10.21037/tbcr-24-36.

Use of Optical Redox Imaging to Quantify Alveolar Macrophage Redox State in Infants: Proof of Concept Experiments in a Murine Model and Human Tracheal Aspirates Samples.

Xu H, Gonzalves D, Hoffman J, Baur J, Li L, Jensen E Antioxidants (Basel). 2024; 13(5).

PMID: 38790651 PMC: 11117937. DOI: 10.3390/antiox13050546.

References

Varone A, Xylas J, Quinn K, Pouli D, Sridharan G, McLaughlin-Drubin M . Endogenous two-photon fluorescence imaging elucidates metabolic changes related to enhanced glycolysis and glutamine consumption in precancerous epithelial tissues. Cancer Res. 2014; 74(11):3067-75. PMC: 4837452. DOI: 10.1158/0008-5472.CAN-13-2713. View

Rehman A, Anwer A, Gosnell M, Mahbub S, Liu G, Goldys E . Fluorescence quenching of free and bound NADH in HeLa cells determined by hyperspectral imaging and unmixing of cell autofluorescence. Biomed Opt Express. 2017; 8(3):1488-1498. PMC: 5480559. DOI: 10.1364/BOE.8.001488. View

Li L, Li W . Epithelial-mesenchymal transition in human cancer: comprehensive reprogramming of metabolism, epigenetics, and differentiation. Pharmacol Ther. 2015; 150:33-46. DOI: 10.1016/j.pharmthera.2015.01.004. View

Heikal A . Intracellular coenzymes as natural biomarkers for metabolic activities and mitochondrial anomalies. Biomark Med. 2010; 4(2):241-63. PMC: 2905054. DOI: 10.2217/bmm.10.1. View

Xu H, Feng M, Nath K, Nelson D, Roman J, Zhao H . Optical Redox Imaging of Lonidamine Treatment Response of Melanoma Cells and Xenografts. Mol Imaging Biol. 2018; 21(3):426-435. PMC: 6525077. DOI: 10.1007/s11307-018-1258-z. View

Wright B, Andrews L, Jones M, Stringari C, Digman M, Gratton E . Phasor-FLIM analysis of NADH distribution and localization in the nucleus of live progenitor myoblast cells. Microsc Res Tech. 2012; 75(12):1717-22. PMC: 3635844. DOI: 10.1002/jemt.22121. View

Feng M, Xu H, Jiang J, Li L . Potential Biomarker for Triple-Negative Breast Cancer Invasiveness by Optical Redox Imaging. Adv Exp Med Biol. 2021; 1269:247-251. PMC: 8172355. DOI: 10.1007/978-3-030-48238-1_39. View

Li L, Zhou R, Xu H, Moon L, Zhong T, Kim E . Quantitative magnetic resonance and optical imaging biomarkers of melanoma metastatic potential. Proc Natl Acad Sci U S A. 2009; 106(16):6608-13. PMC: 2672511. DOI: 10.1073/pnas.0901807106. View

Sun N, Xu H, Luo Q, Li L . Potential Indexing of the Invasiveness of Breast Cancer Cells by Mitochondrial Redox Ratios. Adv Exp Med Biol. 2016; 923:121-127. PMC: 5476445. DOI: 10.1007/978-3-319-38810-6_16. View

10.

Vishwasrao H, Heikal A, Kasischke K, Webb W . Conformational dependence of intracellular NADH on metabolic state revealed by associated fluorescence anisotropy. J Biol Chem. 2005; 280(26):25119-26. DOI: 10.1074/jbc.M502475200. View

11.

White F, Jin Y, Yang L . An evaluation of the role of nuclear cytoplasmic ratios and nuclear volume densities as diagnostic indicators in metaplastic, dysplastic and neoplastic lesions of the human cheek. Histol Histopathol. 1997; 12(1):69-77. View

12.

Wiese M, Bannister A . Two genomes, one cell: Mitochondrial-nuclear coordination via epigenetic pathways. Mol Metab. 2020; 38:100942. PMC: 7300384. DOI: 10.1016/j.molmet.2020.01.006. View

13.

Lim C, Kim E, Kim J, Hong S, Chun H, Kang D . Measurement of the Nucleus Area and Nucleus/Cytoplasm and Mitochondria/Nucleus Ratios in Human Colon Tissues by Dual-Colour Two-Photon Microscopy Imaging. Sci Rep. 2015; 5:18521. PMC: 4682082. DOI: 10.1038/srep18521. View

14.

Mubaid F, Kaufman D, Wee T, Nguyen-Huu D, Young D, Anghelopoulou M . Fluorescence microscope light source stability. Histochem Cell Biol. 2019; 151(4):357-366. DOI: 10.1007/s00418-019-01776-6. View

15.

Sweeney K, Swarbrick A, Sutherland R, Musgrove E . Lack of relationship between CDK activity and G1 cyclin expression in breast cancer cells. Oncogene. 1998; 16(22):2865-78. DOI: 10.1038/sj.onc.1201814. View

16.

Walsh A, Skala M . Optical metabolic imaging quantifies heterogeneous cell populations. Biomed Opt Express. 2015; 6(2):559-73. PMC: 4354590. DOI: 10.1364/BOE.6.000559. View

17.

Dai X, Cheng H, Bai Z, Li J . Breast Cancer Cell Line Classification and Its Relevance with Breast Tumor Subtyping. J Cancer. 2017; 8(16):3131-3141. PMC: 5665029. DOI: 10.7150/jca.18457. View

18.

Chance B, Schoener B, Oshino R, Itshak F, Nakase Y . Oxidation-reduction ratio studies of mitochondria in freeze-trapped samples. NADH and flavoprotein fluorescence signals. J Biol Chem. 1979; 254(11):4764-71. View

19.

Georgakoudi I, Quinn K . Optical imaging using endogenous contrast to assess metabolic state. Annu Rev Biomed Eng. 2012; 14:351-67. DOI: 10.1146/annurev-bioeng-071811-150108. View

20.

Wallrabe H, Svindrych Z, Alam S, Siller K, Wang T, Kashatus D . Segmented cell analyses to measure redox states of autofluorescent NAD(P)H, FAD & Trp in cancer cells by FLIM. Sci Rep. 2018; 8(1):79. PMC: 5758727. DOI: 10.1038/s41598-017-18634-x. View