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Epithelial-to-mesenchymal transition status correlated with ultrastructural features, and TP53 mutation in patient-derived oral cancer cell lines

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Abstract

Background

Oral Squamous Cell Carcinoma (OSCC) is a highly prevalent cancer in the Indian subcontinent. The major cause of mortality in OSCC patients is metastasis. Epithelial-to-mesenchymal transition (EMT) marks an important step in the metastatic process. Additionally, TP53, an important tumor suppressor gene, is also a significant determinant of the treatment outcome, and also plays a role in EMT. Therefore, understanding the interconnections between ultrastructural features, EMT status and TP53 mutational status is of vital importance.

Methods and results

The ultrastructure of five OSCC cell lines was visualized by transmission electron microscopy. Trans-well invasion and migration assays as well as scratch-wound assay, and the expression of various EMT-related genes were utilized to assess the EMT status of the cell lines. The TP53 exons were amplified for the ACOSC3, ACOSC4 and ACOSC16 cell lines and sequenced and the mutations in the gene were identified by sequence alignment. The TP53 mutation in the UPCI:SCC029B cell line has been previously reported, while UPCI:SCC040 has been reported to harbor a wild type TP53. The ACOSC4 cell line which showed the shortest intercellular gaps, also had the least invasive and migratory potential. Interestingly, ACOSC4 showed the highest expression of E-cadherin and the lowest expression of Vimentin, TWIST1, ZEB1, and MMPs. Additionally, TP53 gene of ACOSC4 was unmutated, whereas the ACOSC3 and ACOSC16 harbored TP53 mutations. The mutation in ACOSC3 (R196*) was also found in 7 TCGA samples. Similarly, the UPCI:SCC040 cell line that harbors a wild type TP53 showed shorter intracellular gaps.

Conclusions

Cellular migratory properties are associated with cellular ultrastructure, epithelial-to-mesenchymal transition status and the status of TP53 mutation in the genome.

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Data availability

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank the ACTREC Digital Imaging, Electron Microscopy and Sequencing facilities. We thank Prof Susanne M Gollin, University of Pittsburg, USA for providing the UPCI:SCC029B and UPCI:SCC040 cell lines.

Funding

Sushant S Navarange is supported by Council of Scientific and Industrial Research (CSIR) fellowship. Darshan Mehta is supported by ACTREC fellowship. The present study was supported from the grant of ACTREC-TMC intramural fund (Grant No. 3542) and Department of Atomic Energy (DAE), Government of India (Grant No. 4598).

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Authors and Affiliations

  1. Waghmare Lab, Stem Cell Biology Group, Advanced Centre for Treatment Research and Education in Cancer (ACTREC), Tata Memorial Centre, Cancer Research Institute, Navi Mumbai, 410210, India

    Sushant S. Navarange, Sanjay M. Bane, Darshan Mehta & Sanjeev K. Waghmare

  2. Gupta Lab, Epigenetics and chromatin Biology Group, Advanced Centre for Treatment Research and Education in Cancer (ACTREC), Tata Memorial Centre, Cancer Research Institute, Navi Mumbai, 410210, India

    Sanket Shah & Sanjay Gupta

  3. Training School Complex, Homi Bhabha National Institute, Anushakti Nagar, Mumbai, 400085, India

    Sushant S. Navarange, Darshan Mehta, Sanket Shah, Sanjay Gupta & Sanjeev K. Waghmare

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Contributions

SKW conceived and designed the study. Material preparation, data collection and analysis was performed by SSN, DM, SB, SGS, SKW and SG. SKW reviewed the data and the manuscript. The first draft of the manuscript was written by SSN and SKW. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Sanjeev K. Waghmare.

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Conflict of interest

The authors declare no competing interests.

Ethical approval

All the three cell lines such as ACOSC3, ACOSC4 and ACOSC16, were established from the oral cancer patient samples from Tata Memorial Centre. This study was performed in line with the principles of Helsinki. Approval was granted by the Institutional Human Ethics Committee (Dated: December 14, 2015–IEC/188).

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Informed consent was obtained from all the individual participants included in the study. This study was approved by the Institutional Human Ethics Committee.

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The authors affirm that human research participants provided informed consent for the publication of the images in this manuscript.

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Navarange, S.S., Bane, S.M., Mehta, D. et al. Epithelial-to-mesenchymal transition status correlated with ultrastructural features, and TP53 mutation in patient-derived oral cancer cell lines. Mol Biol Rep 50, 8469–8481 (2023). https://doi.org/10.1007/s11033-023-08720-x

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