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Multigene Mutation Analysis on Cytologic SamplesMultigene Mutation Analysis on Cytologic Samples FREE TO VIEW

Jocelyne Fleury-Feith, MD, PhD; Valérie Gounant, MD; Roger Lacave, MD, PhD; Jacques Cadranel, MD, PhD; Jean-François Bernaudin, MD, PhD
Author and Funding Information

From the Service d’Histologie et Biologie Tumorale (Drs Fleury-Feith, Lacave, and Bernaudin), Service de Pneumologie et Réanimation (Drs Gounant and Cadranel), and Service de Chirurgie Thoracique (Dr Gounant), Hôpital Tenon, AP-HP, Faculté de Médecine, Université Pierre et Marie Curie.

Correspondence to: Jocelyne Fleury-Feith, MD, PhD, Service d’Histologie et Biologie Tumorale, Hôpital Tenon, 4 Rue de la Chine, 75020 Paris, France; e-mail: jocelyne.fleury@tnn.aphp.fr

Financial/nonfinancial disclosures: The authors have reported to CHEST that no potential conflicts of interest exist with any companies/organizations whose products or services may be discussed in this article.

Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (http://www.chestpubs.org/site/misc/reprints.xhtml).


Financial/nonfinancial disclosures: The authors have reported to CHEST that no potential conflicts of interest exist with any companies/organizations whose products or services may be discussed in this article.

Financial/nonfinancial disclosures: The authors have reported to CHEST that no potential conflicts of interest exist with any companies/organizations whose products or services may be discussed in this article.

Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (http://www.chestpubs.org/site/misc/reprints.xhtml).

Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (http://www.chestpubs.org/site/misc/reprints.xhtml).


Chest. 2011;140(6):1664. doi:10.1378/chest.11-1238
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To the Editor:

We read with great interest the article by Nakajima et al1 that was recently published in CHEST (November 2011). The authors demonstrated in a great number of samples that a cytologic specimen obtained by endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) could be of particular interest for a successful gene analysis of epidermal growth factor receptor (EGFR), V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (K-ras), or tumor protein 53 (p53).

Other articles have demonstrated already the feasibility of such an approach for EGFR gene analyses on cytologic specimens,2,3 particularly using paraffin-embedded cell blocks. Moreover, in the recent publication on the classification of lung cancer through small biopsy specimen and cytology examination, Travis et al4 also recommended to systematically use paraffin-embedded cell blocks not only for immunocytochemistry but also for molecular analysis. In contrast, Nakajima et al1 emphasized a very important technical point: Artifacts leading to false gene mutation identification can be obtained from small DNA samples extracted from paraffin-embedded cell blocks. Specimens obtained by EBUS-TBNA often are of small size, and optimal processing is essential for their management; therefore, we agree with Nakajima et al1 that clinicians must pay attention to their treatment. Freezing and storing aliquots of the samples at −80°C in dimethyl sulfoxide according to the same procedure used for cell line preservation is easy to do. Furthermore, doing so enables optimal cell preservation for morphology and a wide range of complementary techniques, such as molecular analyses. In our institution, as already described,5 we systematically freeze a part of the cytologic specimens obtained by EBUS-TBNA at −80°C. From this frozen material, DNA extraction and sequencing usually are performed with success, whatever the specimen cellularity. These techniques are routinely performed not only on samples obtained by EBUS-TBNA, but also on other cytologic samples such as those obtained by transthoracic needle aspiration or bronchial brushings and even cerebrospinal fluid. In conclusion, we agree with Nakajima et al1 that multigene mutation analysis can be performed in EBUS-TBNA samples, promoting freezing cells rather than the cell block and, thus, ensuring optimum technical conditions.

Nakajima T, Yasufuku K, Nakagawara A, Kimura H, Yoshino I. Multigene mutation analysis of metastatic lymph nodes in non-small cell lung cancer diagnosed by endobronchial ultrasound-guided transbronchial needle aspiration. Chest. 2011;1405:1319-1324 [CrossRef] [PubMed]
 
Nicholson AG, Gonzalez D, Shah P, et al. Refining the diagnosis and EGFR status of non-small cell lung carcinoma in biopsy and cytologic material, using a panel of mucin staining, TTF-1, cytokeratin 5/6, and P63, and EGFR mutation analysis. J Thorac Oncol. 2010;54:436-441 [CrossRef]
 
Savic S, Tapia C, Grilli B, et al. Comprehensive epidermal growth factor receptor gene analysis from cytological specimens of non-small-cell lung cancers. Br J Cancer. 2008;981:154-160 [CrossRef]
 
Travis WD, Brambilla E, Noguchi M, et al. International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society international multidisciplinary classification of lung adenocarcinoma. J Thorac Oncol. 2011;62:244-285 [CrossRef]
 
Gounant V, Ninane V, Janson X, et al. Release of metal particles from needles used for transbronchial needle aspiration. Chest. 2011;1391:138-143 [CrossRef]
 

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References

Nakajima T, Yasufuku K, Nakagawara A, Kimura H, Yoshino I. Multigene mutation analysis of metastatic lymph nodes in non-small cell lung cancer diagnosed by endobronchial ultrasound-guided transbronchial needle aspiration. Chest. 2011;1405:1319-1324 [CrossRef] [PubMed]
 
Nicholson AG, Gonzalez D, Shah P, et al. Refining the diagnosis and EGFR status of non-small cell lung carcinoma in biopsy and cytologic material, using a panel of mucin staining, TTF-1, cytokeratin 5/6, and P63, and EGFR mutation analysis. J Thorac Oncol. 2010;54:436-441 [CrossRef]
 
Savic S, Tapia C, Grilli B, et al. Comprehensive epidermal growth factor receptor gene analysis from cytological specimens of non-small-cell lung cancers. Br J Cancer. 2008;981:154-160 [CrossRef]
 
Travis WD, Brambilla E, Noguchi M, et al. International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society international multidisciplinary classification of lung adenocarcinoma. J Thorac Oncol. 2011;62:244-285 [CrossRef]
 
Gounant V, Ninane V, Janson X, et al. Release of metal particles from needles used for transbronchial needle aspiration. Chest. 2011;1391:138-143 [CrossRef]
 
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