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Accuracy of microRNAs as markers for the detection of neck lymph node metastases in patients with head and neck squamous cell carcinoma.

de Carvalho AC, Scapulatempo-Neto C, Maia DC, Evangelista AF, Morini MA, Carvalho AL, Vettore AL - BMC Med (2015)

Bottom Line: Molecular markers based on tissue-specific microRNA expression are alternative accurate diagnostic markers.Seven microRNAs highly expressed in metastatic lymph nodes from the discovery set were validated in FFPE lymph node samples.MiR-203 and miR-205 identified all metastatic samples, regardless of the size of the metastatic deposit.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cancer Molecular Biology, Department of Biological Sciences, Diadema Campus, Federal University of São Paulo, Rua Pedro de Toledo, 669, São Paulo, SP, 04039-032, Brazil. ca_oak@yahoo.com.br.

ABSTRACT

Background: The presence of metastatic disease in cervical lymph nodes of head and neck squamous cell carcinoma (HNSCC) patients is a very important determinant in therapy choice and prognosis, with great impact in overall survival. Frequently, routine lymph node staging cannot detect occult metastases and the post-surgical histologic evaluation of resected lymph nodes is not sensitive in detecting small metastatic deposits. Molecular markers based on tissue-specific microRNA expression are alternative accurate diagnostic markers. Herein, we evaluated the feasibility of using the expression of microRNAs to detect metastatic cells in formalin-fixed paraffin-embedded (FFPE) lymph nodes and in fine-needle aspiration (FNA) biopsies of HNSCC patients.

Methods: An initial screening compared the expression of 667 microRNAs in a discovery set comprised by metastatic and non-metastatic lymph nodes from HNSCC patients. The most differentially expressed microRNAs were validated by qRT-PCR in two independent cohorts: i) 48 FFPE lymph node samples, and ii) 113 FNA lymph node biopsies. The accuracy of the markers in identifying metastatic samples was assessed through the analysis of sensitivity, specificity, accuracy, negative predictive value, positive predictive value, and area under the curve values.

Results: Seven microRNAs highly expressed in metastatic lymph nodes from the discovery set were validated in FFPE lymph node samples. MiR-203 and miR-205 identified all metastatic samples, regardless of the size of the metastatic deposit. Additionally, these markers also showed high accuracy when FNA samples were examined.

Conclusions: The high accuracy of miR-203 and miR-205 warrant these microRNAs as diagnostic markers of neck metastases in HNSCC. These can be evaluated in entire lymph nodes and in FNA biopsies collected at different time-points such as pre-treatment samples, intraoperative sentinel node biopsy, and during patient follow-up. These markers can be useful in a clinical setting in the management of HNSCC patients from initial disease staging and therapy planning to patient surveillance.

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Expression profile of miR-203 and miR-205 in FNA biopsies from lymph nodes classified as positive (FNA+) or negative (FNA–) according to (A) cytological diagnostic (FNA+: n = 42; FNA–; n = 71) and (B) histological diagnostic (FNA+: n = 45; FNA–; n = 68). The Y-axis shows the log10 fold-change of the relative expression (2-ΔΔCt). The P value (Mann–Whitney) from each comparison is provided. The dotted line indicates the 10-fold cutoff adopted.
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Fig6: Expression profile of miR-203 and miR-205 in FNA biopsies from lymph nodes classified as positive (FNA+) or negative (FNA–) according to (A) cytological diagnostic (FNA+: n = 42; FNA–; n = 71) and (B) histological diagnostic (FNA+: n = 45; FNA–; n = 68). The Y-axis shows the log10 fold-change of the relative expression (2-ΔΔCt). The P value (Mann–Whitney) from each comparison is provided. The dotted line indicates the 10-fold cutoff adopted.

Mentions: The accuracy of both markers in detecting metastatic deposits in the FNA biopsies was compared with the results obtained from cytological assessment, which had detected 42 positive FNA samples and 71 negative ones for the presence of epithelial cells. This analysis showed a complete concordance between the cytological and molecular approaches in the detection of positive and negative nodes with a Kappa value of 1.000. All 42 positive and 71 negative FNA on the cytological assessment were correctly identified by both markers with a sensitivity rate of 100% (42/42, CI 95%, 91.5–100) and a specificity level of 100% (71/71, CI 95%, 94.9–100) (Table 2; Figure 6A). A high accuracy of both miR-203 and miR-205 was observed with an AUC of 1.00 (Table 3; Figure 7A), negative and positive predictive values of 100% (95% CI, 94.9–100 and 95% CI, 91.5–100, respectively), and accuracy levels of 100% (95% CI, 96.05–100) (Table 3).Figure 6


Accuracy of microRNAs as markers for the detection of neck lymph node metastases in patients with head and neck squamous cell carcinoma.

de Carvalho AC, Scapulatempo-Neto C, Maia DC, Evangelista AF, Morini MA, Carvalho AL, Vettore AL - BMC Med (2015)

Expression profile of miR-203 and miR-205 in FNA biopsies from lymph nodes classified as positive (FNA+) or negative (FNA–) according to (A) cytological diagnostic (FNA+: n = 42; FNA–; n = 71) and (B) histological diagnostic (FNA+: n = 45; FNA–; n = 68). The Y-axis shows the log10 fold-change of the relative expression (2-ΔΔCt). The P value (Mann–Whitney) from each comparison is provided. The dotted line indicates the 10-fold cutoff adopted.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493814&req=5

Fig6: Expression profile of miR-203 and miR-205 in FNA biopsies from lymph nodes classified as positive (FNA+) or negative (FNA–) according to (A) cytological diagnostic (FNA+: n = 42; FNA–; n = 71) and (B) histological diagnostic (FNA+: n = 45; FNA–; n = 68). The Y-axis shows the log10 fold-change of the relative expression (2-ΔΔCt). The P value (Mann–Whitney) from each comparison is provided. The dotted line indicates the 10-fold cutoff adopted.
Mentions: The accuracy of both markers in detecting metastatic deposits in the FNA biopsies was compared with the results obtained from cytological assessment, which had detected 42 positive FNA samples and 71 negative ones for the presence of epithelial cells. This analysis showed a complete concordance between the cytological and molecular approaches in the detection of positive and negative nodes with a Kappa value of 1.000. All 42 positive and 71 negative FNA on the cytological assessment were correctly identified by both markers with a sensitivity rate of 100% (42/42, CI 95%, 91.5–100) and a specificity level of 100% (71/71, CI 95%, 94.9–100) (Table 2; Figure 6A). A high accuracy of both miR-203 and miR-205 was observed with an AUC of 1.00 (Table 3; Figure 7A), negative and positive predictive values of 100% (95% CI, 94.9–100 and 95% CI, 91.5–100, respectively), and accuracy levels of 100% (95% CI, 96.05–100) (Table 3).Figure 6

Bottom Line: Molecular markers based on tissue-specific microRNA expression are alternative accurate diagnostic markers.Seven microRNAs highly expressed in metastatic lymph nodes from the discovery set were validated in FFPE lymph node samples.MiR-203 and miR-205 identified all metastatic samples, regardless of the size of the metastatic deposit.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cancer Molecular Biology, Department of Biological Sciences, Diadema Campus, Federal University of São Paulo, Rua Pedro de Toledo, 669, São Paulo, SP, 04039-032, Brazil. ca_oak@yahoo.com.br.

ABSTRACT

Background: The presence of metastatic disease in cervical lymph nodes of head and neck squamous cell carcinoma (HNSCC) patients is a very important determinant in therapy choice and prognosis, with great impact in overall survival. Frequently, routine lymph node staging cannot detect occult metastases and the post-surgical histologic evaluation of resected lymph nodes is not sensitive in detecting small metastatic deposits. Molecular markers based on tissue-specific microRNA expression are alternative accurate diagnostic markers. Herein, we evaluated the feasibility of using the expression of microRNAs to detect metastatic cells in formalin-fixed paraffin-embedded (FFPE) lymph nodes and in fine-needle aspiration (FNA) biopsies of HNSCC patients.

Methods: An initial screening compared the expression of 667 microRNAs in a discovery set comprised by metastatic and non-metastatic lymph nodes from HNSCC patients. The most differentially expressed microRNAs were validated by qRT-PCR in two independent cohorts: i) 48 FFPE lymph node samples, and ii) 113 FNA lymph node biopsies. The accuracy of the markers in identifying metastatic samples was assessed through the analysis of sensitivity, specificity, accuracy, negative predictive value, positive predictive value, and area under the curve values.

Results: Seven microRNAs highly expressed in metastatic lymph nodes from the discovery set were validated in FFPE lymph node samples. MiR-203 and miR-205 identified all metastatic samples, regardless of the size of the metastatic deposit. Additionally, these markers also showed high accuracy when FNA samples were examined.

Conclusions: The high accuracy of miR-203 and miR-205 warrant these microRNAs as diagnostic markers of neck metastases in HNSCC. These can be evaluated in entire lymph nodes and in FNA biopsies collected at different time-points such as pre-treatment samples, intraoperative sentinel node biopsy, and during patient follow-up. These markers can be useful in a clinical setting in the management of HNSCC patients from initial disease staging and therapy planning to patient surveillance.

Show MeSH
Related in: MedlinePlus