Viability Assessment Following Anticancer Treatment Requires Single-Cell Visualization

Overview

Journal Cancers (Basel)

Publisher MDPI

Specialty Oncology

Date 2018 Aug 4

PMID 30071623

Citations 14

Authors

Razmik Mirzayans

Bonnie Andrais

David Murray

Affiliations

Soon will be listed here.

Abstract

A subset of cells within solid tumors become highly enlarged and enter a state of dormancy (sustained proliferation arrest) in response to anticancer treatment. Although dormant cancer cells might be scored as "dead" in conventional preclinical assays, they remain viable, secrete growth-promoting factors, and can give rise to progeny with stem cell-like properties. Furthermore, cancer cells exhibiting features of apoptosis (e.g., caspase-3 activation) following genotoxic stress can undergo a reversal process called anastasis and survive. Consistent with these observations, single-cell analysis of adherent cultures (solid tumor-derived cell lines with differing p53 status) has demonstrated that virtually all cells-irrespective of their size and morphology-that remain adherent to the culture dish for a long time (weeks) after treatment with anticancer agents exhibit the ability to metabolize 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl- tetrazolium bromide (MTT). The purpose of this commentary is to briefly review these findings and discuss the significance of single-cell (versus population averaged) observation methods for assessment of cancer cell viability and metabolic activity.

Citing Articles

Single-Cell MTT: A Simple and Sensitive Assay for Determining the Viability and Metabolic Activity of Polyploid Giant Cancer Cells (PGCCs).

Mirzayans R, Andrais B, Murray D Methods Mol Biol. 2024; 2825:293-308.

PMID: 38913317 DOI: 10.1007/978-1-0716-3946-7_17.

Pharmaceutical characterization and exploration of Arkeshwara rasa in MDA-MB-231 cells.

Jayakumar R, Dash M, Kumar P, Sharma S, Gulati S, Pandey A J Ayurveda Integr Med. 2023; 15(1):100823.

PMID: 38160612 PMC: 10792653. DOI: 10.1016/j.jaim.2023.100823.

Intratumor Heterogeneity and Treatment Resistance of Solid Tumors with a Focus on Polyploid/Senescent Giant Cancer Cells (PGCCs).

Mirzayans R, Murray D Int J Mol Sci. 2023; 24(14).

PMID: 37511291 PMC: 10380821. DOI: 10.3390/ijms241411534.

What Are the Reasons for Continuing Failures in Cancer Therapy? Are Misleading/Inappropriate Preclinical Assays to Be Blamed? Might Some Modern Therapies Cause More Harm than Benefit?.

Mirzayans R, Murray D Int J Mol Sci. 2022; 23(21).

PMID: 36362004 PMC: 9655591. DOI: 10.3390/ijms232113217.

The effect of ciprofloxacin on doxorubicin cytotoxic activity in the acquired resistance to doxorubicin in DU145 prostate carcinoma cells.

Davary Avareshk A, Jalal R, Gholami J Med Oncol. 2022; 39(12):194.

PMID: 36071289 DOI: 10.1007/s12032-022-01787-9.

References

Huang Q, Li F, Liu X, Li W, Shi W, Liu F . Caspase 3-mediated stimulation of tumor cell repopulation during cancer radiotherapy. Nat Med. 2011; 17(7):860-6. PMC: 3132290. DOI: 10.1038/nm.2385. View

ONeil N, Bailey M, Hieter P . Synthetic lethality and cancer. Nat Rev Genet. 2017; 18(10):613-623. DOI: 10.1038/nrg.2017.47. View

Karamitopoulou E, Cioccari L, Jakob S, Vallan C, Schaffner T, Zimmermann A . Active caspase 3 and DNA fragmentation as markers for apoptotic cell death in primary and metastatic liver tumours. Pathology. 2007; 39(6):558-64. DOI: 10.1080/00313020701684375. View

Berridge M, Herst P, Tan A . Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnol Annu Rev. 2005; 11:127-52. DOI: 10.1016/S1387-2656(05)11004-7. View

Chen H, Lin F, Xing K, He X . The reverse evolution from multicellularity to unicellularity during carcinogenesis. Nat Commun. 2015; 6:6367. DOI: 10.1038/ncomms7367. View

Feinerman O, Veiga J, Dorfman J, Germain R, Altan-Bonnet G . Variability and robustness in T cell activation from regulated heterogeneity in protein levels. Science. 2008; 321(5892):1081-4. PMC: 2673522. DOI: 10.1126/science.1158013. View

Vincent M . Resistance to cancer chemotherapy as an atavism? (retrospective on DOI 10.1002/bies.201300170). Bioessays. 2016; 38(11):1065. DOI: 10.1002/bies.201600166. View

Liu Y, Peterson D, Kimura H, Schubert D . Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. J Neurochem. 1997; 69(2):581-93. DOI: 10.1046/j.1471-4159.1997.69020581.x. View

Niepel M, Spencer S, Sorger P . Non-genetic cell-to-cell variability and the consequences for pharmacology. Curr Opin Chem Biol. 2009; 13(5-6):556-61. PMC: 2975492. DOI: 10.1016/j.cbpa.2009.09.015. View

10.

Cohen A, Geva-Zatorsky N, Eden E, Frenkel-Morgenstern M, Issaeva I, Sigal A . Dynamic proteomics of individual cancer cells in response to a drug. Science. 2008; 322(5907):1511-6. DOI: 10.1126/science.1160165. View

11.

Irish J, Hovland R, Krutzik P, Perez O, Bruserud O, Gjertsen B . Single cell profiling of potentiated phospho-protein networks in cancer cells. Cell. 2004; 118(2):217-28. DOI: 10.1016/j.cell.2004.06.028. View

12.

Perez-Garijo A . When dying is not the end: Apoptotic caspases as drivers of proliferation. Semin Cell Dev Biol. 2017; 82:86-95. DOI: 10.1016/j.semcdb.2017.11.036. View

13.

Batchelor E, Loewer A . Recent progress and open challenges in modeling p53 dynamics in single cells. Curr Opin Syst Biol. 2017; 3:54-59. PMC: 5650191. DOI: 10.1016/j.coisb.2017.04.007. View

14.

Esmatabadi M, Bakhshinejad B, Motlagh F, Babashah S, Sadeghizadeh M . Therapeutic resistance and cancer recurrence mechanisms: Unfolding the story of tumour coming back. J Biosci. 2016; 41(3):497-506. DOI: 10.1007/s12038-016-9624-y. View

15.

Husmann M . Vital dyes and virtual deaths. Cell Death Differ. 2013; 20(7):963. PMC: 3679458. DOI: 10.1038/cdd.2013.27. View

16.

Saadatpour A, Lai S, Guo G, Yuan G . Single-Cell Analysis in Cancer Genomics. Trends Genet. 2015; 31(10):576-586. PMC: 5282606. DOI: 10.1016/j.tig.2015.07.003. View

17.

Elmore S . Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007; 35(4):495-516. PMC: 2117903. DOI: 10.1080/01926230701320337. View

18.

Albeck J, Burke J, Aldridge B, Zhang M, Lauffenburger D, Sorger P . Quantitative analysis of pathways controlling extrinsic apoptosis in single cells. Mol Cell. 2008; 30(1):11-25. PMC: 2858979. DOI: 10.1016/j.molcel.2008.02.012. View

19.

Rosenthal K, Oehling V, Dusny C, Schmid A . Beyond the bulk: disclosing the life of single microbial cells. FEMS Microbiol Rev. 2017; 41(6):751-780. PMC: 5812503. DOI: 10.1093/femsre/fux044. View

20.

Mirzayans R, Andrais B, Kumar P, Murray D . Significance of Wild-Type p53 Signaling in Suppressing Apoptosis in Response to Chemical Genotoxic Agents: Impact on Chemotherapy Outcome. Int J Mol Sci. 2017; 18(5). PMC: 5454841. DOI: 10.3390/ijms18050928. View