Abstract
Papillary thyroid carcinoma (PTC), a common endocrine malignancy, presents a challenge from a prognostic standpoint. Molecular alterations underlying PTC progression include deregulation of focal adhesion kinase (FAK) at post-transcriptional and post-translational levels. Searching for candidate markers of PTC progression, we investigated the prognostic significance of FAK alterations on mRNA/protein level. The expression levels and subcellular localisation of auto-phosphorylated FAK (pY397-FAK) were determined by western blot (WB) and immunohistochemistry. The quantity of total FAK mRNA, alternatively spliced FAK-Del26 and FAK-Del33 variants were analysed by RT-qPCR and related to pY397-FAK expression and subcellular distribution. The results were correlated with clinicopathological parameters of the patients. The expression of pY397-FAK was significantly elevated in malignant samples. Active FAK showed predominant cytoplasmic distribution with co-occurrence along the membrane, while nuclear staining was found less frequently. Expression of pY397-FAK in separate cellular compartments correlated with adverse clinicopathological parameters, but the strongest association was found when their mean scores were calculated. Alternatively spliced FAK-Del33 and total FAK transcripts positively correlated to pY397-FAK protein levels as well as to characteristics of PTC advancement. Over-expression of FAK on mRNA (total and Del-33) and activated protein (pY397-FAK) levels is a feature of PTC advanced stages. Of the analysed alterations, the mean pY397-FAK IHC score showed the best predictive performance. Correlation between mRNA FAK-Del33 and pY397-FAK expression implies a regulatory role of alternative splicing in PTC patients.
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The data sets generated during the current study are available from the corresponding author upon reasonable request.
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Acknowledgements
The authors would like to express their gratitude to Dr Dubravka Cvejić for the critical review of the paper and guidance and support throughout of the study. This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, Agreement No 451-03-9/2021-14/ 200019.
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This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, Agreement No451-03-9/2021-14/ 200019.
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Ignjatović, V.B., Janković Miljuš, J.R., Rončević, J.V. et al. Focal adhesion kinase splicing and protein activation in papillary thyroid carcinoma progression. Histochem Cell Biol 157, 183–194 (2022). https://doi.org/10.1007/s00418-021-02056-y
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DOI: https://doi.org/10.1007/s00418-021-02056-y