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Transcription factor SOX3 upregulated pro-apoptotic genes expression in human breast cancer

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Abstract

Background

Sex-determining region Y-box 3 (SOX3) protein, a SOX transcriptions factors group, has been identified as a key regulator in several diseases, including cancer. Downregulation of transcriptions factors in invasive ductal carcinoma (IDC) can interfere in neoplasia development, increasing its aggressiveness. We investigated SOX3 protein expression and its correlation with apoptosis in the MDA-MB-231 cell line, as SOX3 and Pro-Caspase-3 immunoexpression in paraffin-embedded invasive ductal carcinoma tissue samples from patients (n = 27). Breast cancer cell line MDA-MD-231 transfected with pEF1-SOX3 + and pEF1-Empty vector followed by cytotoxicity assay (MTT), Annexin-V FITC PI for apoptosis percentage assessment by flow cytometry, qPCR for apoptotic-related gene expression, immunofluorescence, and immunohistochemistry to SOX3 immunolocalization in culture cells, and paraffin-embedded invasive ductal carcinoma tissue samples.

Results

Apoptotic rate was higher in cells transfected with pEF1-SOX3 + (56%) than controls (10%). MDA-MB-231 transfected with pEF1-SOX3 + presented upregulation of pro-apoptotic mRNA from CASP3, CASP8, CASP9, and BAX genes, contrasting with downregulation antiapoptotic mRNA from BCL2, compared to non-transfected cells and cells transfected with pEF1-empty vector (p < 0.005). SOX3 protein nuclear expression was detected in 14% (4/27 cases) of ductal carcinoma cases, and pro-Caspase-3 expression was positive in 50% of the cases.

Conclusion

Data suggest that SOX3 transcription factor upregulates apoptosis in breast cancer cell line MDA-MB-231, and has a down nuclear expression in ductal carcinoma cases, and need to be investigated as a tumor suppressor protein, and its loss of expression and non-nuclear action turn the cells resistant to apoptosis. Further studies are necessary to understand how SOX3 protein regulates the promoter regions of genes involved in apoptosis.

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Data availability

Not applicable.

Abbreviations

TFs:

Transcription factors

IDC:

Invasive ductal carcinoma

HMG:

High-mobility group

EMT:

Epithelial mesenchymal transition

IHC:

Immunohistochemistry

DMEM:

Dulbecco’s modified eagle medium

PBS:

Phosphate buffered saline

GFP:

Green fluorescent protein

BSA:

Bovine Serum Albumin

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

DMSO:

Dimethyl sulfoxide

PI:

Propidium iodide

UFMS:

Federal university of mato grosso do sul

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Acknowledgements

We thank the patients and volunteers who contributed to this research. We thank Doctor Amy Milsted very much for reviewing the manuscript.

Funding

Research supported by Federal University of Minas Gerais (Edital PRPq05/20160); This study was part of a PhD thesis by F.H.H.S. at the Pathology Post-Graduation Program at Universidade Federal de Minas Gerais, PhD scholarship funded by CAPES.

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Authors and Affiliations

  1. Department of General Pathology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Campus UFMG, Belo Horizonte, MG, 31270-901, Brazil

    Felipe Henrique de Souza Silva, Camila Pereira Almeida, Thais Salviana Ribeiro, Isabella Terra Borges, Geovanni Dantas Cassali, Enio Ferreira & Helen Lima Del Puerto

  2. Division of Mathematics and Sciences, Walsh University, North Canton, OH, USA

    Adam Underwood

  3. Department of Physiology and Biophysics, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Minas Gerais, Brazil

    Elaine M. Souza-Fagundes, Almir S. Martins, Marcos Eliezeck & Silvia Guatimosim

  4. Department of Morphology, Institute of Biological Sciences, Universidade Federal de Minas Gerais, Minas Gerais, Brazil

    Luciana O. Andrade, Luisa Rezende, Hipacia Werneck Gomes & Cleida Aparecida Oliveira

  5. Program in Health and Development in the Midwest Region of UFMS, Campo Grande, MS, Brazil

    Raquel Cristina Rodrigues

Authors
  1. Felipe Henrique de Souza Silva
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Contributions

FHSS: data curation, formal analysis, investigation, writing—original draft. AU: resources, supervision, writing—review and editing. CPA: data curation, formal analysis, investigation, writing—review and editing. TSR: data curation, investigation, writing—review and editing. EMSF: resources, investigation. ASM: resources, investigation. ME: methodology. SG: methodology, resources. LOA: data curation, resources, methodology. LR: data curation, methodology. HWG: data curation, methodology. CAO: resources, methodology. RCR: patient’s recruitment and sample curation. ITB: data curation, investigation, writing—review and editing. GDC: resources, investigation. EF: data curation, formal analysis, resources, supervision. HLDP: Conceptualization, formal analysis, funding acquisition, methodology, project administration, resources, supervision, writing -review and editing.

Corresponding author

Correspondence to Helen Lima Del Puerto.

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The authors disclose no potential conflicts of interest.

Ethical approval and consent to participate

Informed consent was obtained from all the subjects before inclusion in the study, which was approved by the local Human Investigation Committee (CAAE 20636419.2.0000.0021).

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Silva, F.H.d., Underwood, A., Almeida, C.P. et al. Transcription factor SOX3 upregulated pro-apoptotic genes expression in human breast cancer. Med Oncol 39, 212 (2022). https://doi.org/10.1007/s12032-022-01758-0

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