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Breast tumour cell subpopulations with expression of the MYC and OCT4 proteins

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Journal of Molecular Histology Aims and scope Submit manuscript

Abstract

The MYC and OCT4 genes are known factors associated with maintaining pluripotency and are linked with a more aggressive course, progression, and resistance to therapy in cancer. Determining the subpopulations of tumour cells expressing the Myc and Oct4 proteins will provide an opportunity to understand which tumour cell subpopulations expressing MYC and OCT4 are associated with metastasis and resistance and which subpopulations can be targeted by anti-MYC and anti-OCT4 therapy. The study included paraffin-embedded tissue from tumours from 27 patients with luminal B breast cancer obtained after neoadjuvant chemotherapy (NACT). Immunofluorescence staining was used to identify subpopulations of tumour cells expressing Myc, Oct4 and Snai2 (Opal™ 7-Color Kit (PerkinElmer, Hopkinton, MA). The following tumour cell subpopulations were identified with the Myc and Oct4 proteins and the Snai2 EMT marker: stem/progenitor tumour cells with/without Myc, Oct4 or Snai2 expression; differentiated tumour cells with/without Myc, Oct4 or Snai2 expression; and other nontumour cells (CK7−EpCAM−CD44+/−Myc+/−(Oct4, Snai2)+/−) within the inflammatory infiltrate in the tumour parenchyma and stroma. The circulating tumour cell subpopulations with Oct4 protein expression in the bloodstream were studied by flow cytometry. It was found that in patients with partial regression (PR) in response to NACT, the frequency of tumour stem cells was 3.6-fold increased (p = 0.038) in the non-EMT state (CK7+EpCam+CD44+Snai2−). In patients with metastases, there was a statistically significant 2.5-fold increase in the frequency of differentiated tumour cells with Myc expression (CK7+EpCam+CD44−Myc+) and a 2.7-fold increase in the frequency of cells with Oct4 expression (CK7+EpCam+CD44−OCT4+). In the next stage, the frequencies of subpopulations with expression of the Oct4 protein and signs of EMT among circulating tumour cells (CTCs) were determined. In patients with metastases, the frequency of tumour stem cells in the EMT state (CD326+CD44+CD24−CD325+) (p = 0.015) was more than fourfold increased, and the frequency of progenitor tumour cells with expression of the Oct4 stem protein (CD326+CD44+CD24+Oct4+) (p = 0.016) was almost sixfold higher than that in patients without metastases. Nonstem (differentiated) tumour cells with expression of the stemness proteins Myc and Oct4 were present in the breast tumour. Their content was significantly higher in residual tumours after NACT in patients who subsequently developed metastases compared with that in patients without metastases. Such cells are a new in situ marker of metastasis.

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Abbreviations

AC:

Adriamycin and cyclophosphamide

AT/ACT:

Adriamycin and taxotere or Adriamycin, cyclophosphamide and taxotere

CSCs:

Cancer stem cells

CTCs:

Circulating tumour cells

EMT:

Epithelial-mesenchymal transition

IL6:

Interleukin 6

NACTP:

Neoadjuvant chemotherapy

pCR:

Pathological complete response

PD:

Progressive disease.

PR:

Partial response

SD:

Stable disease

TCGA:

The Cancer Genome Atlas

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Funding

The study was supported by the Russian Foundation for Basic Research, Grant No. 18–29-09131.

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Authors and Affiliations

  1. Laboratory of Oncovirology, Cancer Research Institute Tomsk NRMC, Tomsk, Russian Federation

    N. V. Litviakov, V. A. Bychkov, M. K. Ibragimova, M. M. Tsyganov & K. A. Gaptulbarova

  2. Biological Institute of National Research Tomsk State University, Tomsk, Russia

    N. V. Litviakov, M. K. Ibragimova & K. A. Gaptulbarova

  3. Laboratory of Molecular Oncology and Immunology, Cancer Research Institute Tomsk NRMC, Tomsk, Russian Federation

    M. N. Stakheeva

  4. Department of General and Molecular Pathology, Cancer Research Institute Tomsk NRMC, Tomsk, Russian Federation

    L. A. Tashireva & L. N. Bondar

  5. Department of General Oncology, Cancer Research Institute Tomsk NRMC, Tomsk, Russian Federation

    E. Yu. Garbukov & E. M. Slonimskaya

  6. Department of Oncology, Faculty of Medicine, Saint-Petersburg State University, Saint-Petersburg, Russian Federation

    E. M. Slonimskaya

Authors
  1. N. V. Litviakov
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  2. V. A. Bychkov
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  3. M. N. Stakheeva
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  4. M. K. Ibragimova
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  5. M. M. Tsyganov
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  6. K. A. Gaptulbarova
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  7. L. A. Tashireva
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  8. L. N. Bondar
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  9. E. Yu. Garbukov
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  10. E. M. Slonimskaya
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Contributions

NVL—general management and the formation of research design, material analysis and article writing. VAB—Identification of subpopulations of tumor cells expressing Myc and Oct4 proteins in situ; MNS—Study of the content of circulating tumor cells with Oct4 expression in the bloodstream. MKI—Keeping bank of biological material; Identification of subpopulations of tumor cells expressing Myc and Oct4 proteins in situ; MMT—Keeping bank of biological material; Identification of subpopulations of tumor cells expressing Myc and Oct4 proteins in situ; KAG—Identification of subpopulations of tumor cells expressing Myc and Oct4 proteins in situ. LAT—Interpretation of the results of morphological analysis. LNB—Interpretation of the results of morphological analysis. EYG—treatment of patients included in research. EMS—clinical research management, material analysis.

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Correspondence to N. V. Litviakov.

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The authors have no conflicts of interest. All authors express the lack of competing interests.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The study was conducted with a permission by the local ethics committee of the Cancer Research Institute Tomsk NRMC (Protocol 1 from January 14, 2013). All patients signed an informed consent. The experimental protocols were approved by a institutional committee Tomsk National Research Medical Center of the Russian Academy of Sciences (Protocol 3 from January 16, 2013).

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Litviakov, N.V., Bychkov, V.A., Stakheeva, M.N. et al. Breast tumour cell subpopulations with expression of the MYC and OCT4 proteins. J Mol Hist 51, 717–728 (2020). https://doi.org/10.1007/s10735-020-09917-1

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  • DOI: https://doi.org/10.1007/s10735-020-09917-1

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