Yonsei Med J.  2019 Nov;60(11):1005-1012. 10.3349/ymj.2019.60.11.1005.

Methionyl-tRNA Synthetase is a Useful Diagnostic Marker for Lymph Node Metastasis in Non-Small Cell Lung Cancer

• Lee JM
• Kim T
• Kim EY
• Kim A
• Lee DK
• Kwon NH
• Kim S
• Chang YS

• KMID: 2460216
• DOI: http://doi.org/10.3349/ymj.2019.60.11.1005

Abstract

PURPOSE
Identification of lymph node (LN) metastasis in non-small cell lung cancer (NSCLC) is critical for disease staging and selection of therapeutic modalities. Sometimes it is not possible to obtain LN core tissue by endobronchial ultrasound-guided transbronchial needle aspirate (EBUS-TBNA), resulting in low diagnostic yield.
MATERIALS AND METHODS
In this study, 138 specimens were collected from 108 patients who underwent EBUS-TBNA under the suspicion of LN metastasis of NSCLC. Diagnostic yields of anti-CD45 and anti-methionyl-tRNA synthetase (MRS), immunofluorescent (IF) staining on cytology specimens were compared with those of conventional cytology and positron emission tomography-computed tomography (PET-CT).
RESULTS
MRS was strongly expressed in NSCLC cells metastasized to LNs, but weakly expressed in cells at the periphery of the LN germinal center. The majority of cells were CD20 positive, although a few cells were either CD3 or CD14 positive, indicating that CD45 staining is required for discrimination of non-malignant LN constituent cells from NSCLC cells. When the diagnostic efficacy of MRS/CD45 IF staining was evaluated using 138 LN cellular aspirates from 108 patients through EBUS-TBNA, the sensitivity was 76.7% and specificity was 90.8%, whereas those of conventional cytology test were 71.8% and 100.0%, respectively. Combining the results of conventional cytology testing and those of PET-CT showed a sensitivity and specificity of 71.6% and 100%, and the addition of MRS/CD45 dual IF data to this combination increased sensitivity and specificity to 85.1% and 97.8%, respectively.
CONCLUSION
MRS/CD45 dual IF staining showed good diagnostic performance and may be a good tool complementing conventional cytology test for determining LN metastasis of NSCLC.

Keyword

Aminoacyl tRNA synthetase; biomarker; EBUS-TBNA; MRS; NSCLC

MeSH Terms

Amino Acyl-tRNA Synthetases
Carcinoma, Non-Small-Cell Lung*
Complement System Proteins
Discrimination (Psychology)
Electrons
Germinal Center
Humans
Ligases
Lymph Nodes*
Methionine-tRNA Ligase*
Needles
Neoplasm Metastasis*
Sensitivity and Specificity
Amino Acyl-tRNA Synthetases
Complement System Proteins
Ligases
Methionine-tRNA Ligase

Figure

• 

  Fig. 1 Hematoxylin and eosin stain (A) and methionyl-tRNA synthetase immunohistochemistry (B and C) staining of three lymph node specimens of non-small cell lung cancer metastasis. Each column represents the same case (A and B: ×100, C: ×400). Representative cases are shown.

• 

  Fig. 2 Immunohistochemistry for methionyl-tRNA synthetase (MRS), CD20 (B-lymphocytes), CD3 (T-lymphocytes), CD14 (macrophages and monocytes), and CD45 (leukocyte common antigen) markers in the lymph nodes with non-small cell lung cancer, splenic tissue, thymic tissue from the thymoma, and tonsils. Magnification: ×100.

• 

  Fig. 3 Immunoblotting of methionyl-tRNA synthetase (MRS)-positive H460 cells, CD45/CD3-positive Molt-4 cells, and CD45/CD20-positive Daudi cells (A). Fluorescent immunochemical staining for MRS and CD45 in the reference samples containing a fixed ratio of H460, Daudi, and Molt-4 cells (B and C) and endobronchial ultrasound-guided transbronchial needle aspirate-derived samples (D and E). Images were taken using a fluorescent microscope (B and D) and confocal microscope (C and E). Blue represents nucleus (4′, 6-Diamidine-2′-phenylindole dihydrochloride: DAPI); green, MRS; and red, CD45. B and D confocal images show merged images only. TTF-1, thyroid transcription factor 1.

• 

  Fig. 4 Receiver operating characteristic (ROC) curves of predictors of lung cancer lymph node metastasis. ROC curves of (A) cytology, (B) methionyl-tRNA synthetase (MRS)/CD45 immunofluorescence (IF), and (C) the combination of cytology, positron emission tomography (PET), and MRS/CD45 IF were created using core biopsy results as a reference value. lr.eta, linear predictor eta; PV(+), positive predictive value; PV(−), negative predictive value; s.e., standard error; est., estimates.

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