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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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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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.
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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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DOI: https://doi.org/10.1007/s12032-022-01758-0