LIMD2 Is Overexpressed in BRAF V600E-Positive Papillary Thyroid Carcinomas and Matched Lymph Node Metastases
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We previously described that LIM domain containing 2 (LIMD2) overexpression was closely correlated with metastatic process in papillary thyroid carcinoma (PTC). We here evaluated the expression of LIMD2 in a series of non-metastatic and metastatic PTC and their matched lymph node metastases via immunohistochemistry. LIMD2 was expressed in 74 (81%) of primary PTC and 35 (95%) of lymph node metastases. Sub-analysis performed in 37 matched samples demonstrated that in four cases, LIMD2 is expressed in lymph node metastases, while it is not expressed in primary tumors. Moreover, in eight cases, the staining intensity of LIMD2 was stronger in the patient-matched lymph node metastases than in the primary tumors. Next, the expression of LIMD2 was correlated with clinical pathological parameters and BRAF V600E and RET/PTC mutational status. The expression of LIMD2 in primary tumors was correlated with the presence of BRAF V600E mutation (P = 0.0338). Western blot analysis in thyroid cell lines demonstrated that LIMD2 is expressed in two PTC cell lines, while it is not expressed in normal thyroid and follicular thyroid carcinoma cell lines. Importantly, its expression was higher in a PTC cell line that harbors BRAF V600E mutation than in a PTC cell line that harbors RET/PTC1. The available genomic profiling data generated by The Cancer Genome Atlas Research Network confirmed that LIMD2 expression is higher in BRAF-like PTC samples. Our data suggest that LIMD2 may play an important role in the metastatic process of PTC, predominantly in BRAF V600E-positive tumors.
KeywordsLIMD2 Papillary thyroid cancer Metastases BRAF V600E and RET/PTC
M.J.C.P.S. performed IHC staining, BRAF V600E mutational status, analyzed and interpreted data, and wrote the manuscript draft. A.U.B. performed BRAF and RET/PTC mutational screening and Western blot analysis, analyzed and interpreted data, prepared all figures, and wrote the manuscript. V.R.C. performed RNA extraction and helped with the RET/PTC screening. R.D. revised all slides and analyzed and interpreted the IHC data. S.C.L. revised the clinical data from all patients. G.A.C.G. helped with the BRAF mutational analysis. H.P. and F.R.J. produced the antibodies, interpreted the data, and revised the manuscript. G.O. was responsible for the sample selection, analyzed and interpreted data, and revised the manuscript. J.M.C. designed the experiments, analyzed the data, revised the manuscript critically for important intellectual content, edited the manuscript, and coordinated the study. All authors reviewed the results and approved the final version of the manuscript.
The study was supported by research grants (2014/06570-6) from The São Paulo State Research Foundation (FAPESP) and grant (470441/2013-5) from The Brazilian Research Council (CNPq). A.U.B. (2012/06221-6), G.O. (2012/17545-7), and V.R.C. (2016/25127-1) are FAPESP scholars. J.M.C. is an investigator with the CNPq.
Compliance with Ethical Standards
The study was conducted under the approval of the Review Boards and Research Ethical Committees of UNIFESP (1309/11).
Conflict of Interest
The authors declare that they have no conflict of interest.
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