LIMD2 Is Overexpressed in BRAF V600E-Positive Papillary Thyroid Carcinomas and Matched Lymph Node Metastases

Abstract

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.

LIM domain-containing 2 (LIMD2) promotes the progress of ovarian cancer via the focal adhesion signaling pathway

“…As a LIM domain protein, LIMD2 has been functionally associated with cell motility and adhesion, mostly by regulating mRNA transcription [ 9 , 31 ]. In our study, we found that LIMD2 was upregulated in OC tissues, which was consistent with the findings in lung cancer [ 11 ] and papillary thyroid cancer [ 10 ]. LIMD2 expression levels were positively correlated with cell motility metastatic potential and grade in both fresh and archived tumors, including bladder melanoma breast and thyroid tumors [ 13 ].…”

“…In addition, LIMD2 plays an important role in tumorigenesis and development. It has been reported that LIMD2 was a lymph node metastasis marker for papillary thyroid carcinoma (PTC) [ 10 ]. Also, LIMD2 was found to be overexpressed in most metastatic lymph nodes in PTC, while it was undetectable or expressed at relatively low levels in the primary PTCs [ 7 ].…”

Revised criteria for diagnosis of NIFTP reveals a better correlation with tumor biological behavior

“…For amplification of exon 15 of BRAF , the PCR protocol was performed as previously described: an initial denaturation step at 94°C for 5 min followed by 40 cycles of a three-step program, namely, 94°C for 30 s, 53°C for 40 s and 72°C for 30 s, and a final 2-min extension at 72°C (37). For amplification of exon 2 of NRAS or HRAS , the amplification protocol was an initial denaturation step at 94°C for 5 min followed by 40 cycles of a three-step program, namely, 94°C for 30 s, 56°C for 45 s and 72°C for 45 s, and a final 7-min extension at 72°C.…”

Machine learning algorithm improved automated droplet classification of ddPCR for detection of BRAF V600E in paraffin-embedded samples

“…Hence a total of 115 thyroid specimens were analysed. As controls, we additionally included FFPE from thyroid samples that had previously been confirmed as positive (PTC) and negative (FTA) for BRAF V600E mutation by Sanger Sequencing 20 – 22 . The study was conducted under the approval of the Research Ethical Committee of the Universidade Federal de São Paulo, São Paulo, Brazil.…”

“…All samples with less than 20% abundance of the BRAF V600E mutated allele by pyrosequencing were screened by sequenced by Sanger Sequencing as previously described 20 – 22 . Additionally, we randomly selected samples with > 20% of abundance of mutated alleles for Sanger Sequencing.…”