Histopathology 2015, 66, 816–823. DOI: 10.1111/his.12516

Novel use for an EGFR mutation-specific antibody in discriminating lung adenocarcinoma from reactive pneumocyte hyperplasia Tomoyasu Mimori,1,2 Saori Kobayashi,1 Ayako Tanaka,1,2 Shinji Sasada,2 Akihiko Yoshida,1 Takehiro Izumo,2 Naoshi Sasaki,1 Takaaki Tsuchida2 & Koji Tsuta1 1

Division of Pathology and Clinical Laboratories, National Cancer Center Hospital, Tokyo, Japan, and 2Division of Endoscopy, Respiratory Endoscopy, National Cancer Research Institute, Tokyo, Japan

Date of submission 26 March 2014 Accepted for publication 22 July 2014 Published online Article Accepted 26 July 2014

Mimori T, Kobayashi S, Tanaka A, Sasada S, Yoshida A, Izumo T, Sasaki N, Tsuchida T & Tsuta K (2015) Histopathology 66, 816–823. DOI: 10.1111/his.12516

Novel use for an EGFR mutation-specific antibody in discriminating lung adenocarcinoma from reactive pneumocyte hyperplasia. Aims: Pulmonary ground-glass nodules (GGNs) are frequently observed. Histopathologically, their presentation can indicate a wide range of disorders from an inflammatory process to malignancy. An accurate diagnosis based on GGNs can sometimes be challenging on small-sized biopsies. Mutations in the EGFR gene are detected in pulmonary adenocarcinomas (ADCs). Immunohistochemical analysis using antibodies that detect specific EGFR mutations has been shown to correlate with mutational status as determined by molecular methods. We hypothesized that these antibodies could be used to discriminate between ADCs and benign pneumocyte hyperplasias. Methods and results: Surgically resected, pre-invasive to invasive lung ADC (n = 32) and reactive pneumocyte hyperplasia (n = 40) tissue samples were probed with

antibodies against EGFR mutations, p53, Mouse double minute 2 and 14-3-3 sigma. Of the 32 lung ADC specimens analysed, 12 (38%) were positive using the EGFR mutation-specific antibodies, while no immunoreactivity was observed in reactive pneumocyte hyperplasia specimens. Analyses of receiver operating characteristic curves showed that the highest area under the curve values were associated with the use of EGFR mutation-specific antibodies. In addition, a high concordance rate was observed between surgically resected and corresponding biopsy materials using these antibodies. Conclusions: EGFR mutation-specific antibodies can be used to discriminate between lung ADC and benign pneumocyte hyperplasia, even in small-sized biopsies.

Keywords: EGFR, immunohistochemistry, lung adenocarcinoma, pneumocyte hyperplasia

Introduction Since the introduction of low-dose helical computed tomography for lung cancer screening, pulmonary ground-glass nodules (GGNs) have frequently been detected.1 A GGN is defined as a hazy, opaque area of increased attenuation with preserved bronchial and Address for correspondence: K Tsuta, Division of Pathology and Clinical Laboratories, National Cancer Center Hospital, 1-1 Tsukiji 5chome, Chuo-ku, Tokyo 104-0045, Japan. e-mail: [email protected] © 2014 John Wiley & Sons Ltd.

vascular margins.2 Their presence can result from various factors, including partial filling of air spaces, interstitial thickening, partial collapse of alveoli or increased capillary blood volume.3 In lung adenocarcinoma (ADC), GGNs can correspond to a tissue area replaced by the growth of tumour cells accompanied by interstitial thickening or the presence of collapse.4,5 In contrast, in benign cases, pneumocyte swelling with thickening of alveolar walls due to inflammatory cell infiltration and/or accumulation in the alveolar space can also manifest as GGNs.6

EGFR immunochemistry discriminating adenocarcinoma

Histological diversity (from inflammation to malignancy) has also been reported among patients with GGN.7,8 When the lesion exhibits a typical or specific morphology, diagnosis is straightforward. However, in cases where there is an overlap in morphological and cytological features, achieving an accurate diagnosis based on small-sized biopsies obtained by limited invasive procedures can be challenging.7 Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase from the ErbB family of transmembrane glycoproteins. Mutations in the EGFR gene are associated with several pathological processes, including increased cell survival and tumour growth,9 and have been detected frequently in pulmonary ADCs, especially in cases with a lepidic component.10 Approximately 90% of EGFR mutations occur at two hotspots: in-frame deletions in exon 19 centred around codons E746_A750, and a point mutation at codon 858 (L858R) in exon 21.11 Recently, some research groups have reported that immunohistochemistry (IHC) results obtained using EGFR mutation-specific antibodies correlate well with mutational status, as determined by molecular methods.12–15 Unlike molecular-based assays that provide information on the whole tissue, IHC has the advantage of allowing the detailed visualization of protein distribution in situ by light microscopy. We hypothesized that IHC using EGFR mutationspecific antibodies could enable discrimination between pre-invasive to invasive lung ADC and benign pneumocyte hyperplasia. The diagnostic utility of p53, Mouse double minute 2 (MDM2) or 14-3-3 sigma (stratifin: SFN) antibodies was also assessed, because higher expression levels of these proteins have been reported in ADC.16,17

Materials and methods

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ological characteristics are shown in Table S1. In addition, there were 40 cases of pneumocyte hyperplasia induced by benign diseases of the lung, including granulomatous inflammation (n = 33), non-granulomatous chronic inflammation (n = 5), organizing pneumonia (n = 1) and bacterial infection (n = 1). Samples were formalin-fixed and embedded in paraffin blocks that were sectioned at a thickness of 4 lm. IMMUNOHISTOCHEMISTRY AND SAMPLE EVALUATION

For immunohistochemical staining, tissue sections were deparaffinized according to standard protocols. Details of the antigen retrieval method and dilutions for each primary antibody are listed in Table 1. An automated stainer (Dako, Carpinteria, CA, USA) was used according to the manufacturer’s protocol. Positive staining for SFN and MDM2 was defined as more than 10% of cells exhibiting distinct nuclear staining; and with both EGFR mutation-specific antibodies as more than 10% of cells exhibiting distinct cytoplasmic or membrane staining, immunoreactivity to either of these (L858R or E746_A750) being counted as positive. p53-positive cases were defined using two thresholds: 30% and 50% of cells with distinct nuclear staining. ANALYSES OF THE EGFR MUTATIONAL STATUS

In the ADC cases, two common EGFR mutations, namely deletions in exon 19 (DEL) and a point mutaTable 1. Summary of antibodies

Antibody

Clone

EGFR Del E746_A750

6B6

EGFR L858R

Antigen retrieval

Dilution

Source

TRS9 (DAKO)

1:200

Cell Signaling

43B2

TRS9 (DAKO)

1:200

Cell Signaling

p53

DO-7

Citrate buffer

1:100

DAKO

MDM2

1F2

TRS9 (DAKO)

1:400

Invitrogen

Anti-Human 14-3-3 r Protein (N)

18646

TRS9 (DAKO)

1:50

PATIENTS AND SAMPLES

The study was approved by the institutional review board (2010-0077). All available slides for each case were examined, and histopathological subtyping was performed according to the IASLC/ATS/ERS International Multidisciplinary Classification of lung ADC.18 The cohort consisted of 32 cases of resected, pre-invasive to invasive lung ADC, including three cases of adenocarcinoma in situ (AIS) and five minimally invasive adenocarcinomas (MIA), as well as 19 lepidic, three papillary and two acinar predominant ADCs, in which GGNs were identified by radiography. There were no mucinous AIS or MIA cases (formerly mucinous bronchioloalveolar carcinoma). Clinical and path© 2014 John Wiley & Sons Ltd, Histopathology, 66, 816–823.

ImmunoBiological Laboratories, Gunma, Japan

EGFR, Epidermal growth factor receptor; MDM2, Mouse double minute 2.

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tion at codon 858 in exon 21 (L858R), were detected in a high-resolution melting analysis.

Table 2. Results of immunohistochemistry using each antibody in adenocarcinomas and benign diseases

STATISTICAL ANALYSIS

IHC

ADCs (n = 32) (%)

Statistical analysis was performed using SPSS statistics version 21 (IBM Corporation, Somers, NY, USA). The area under the curve (AUC) in receiver operating characteristics (ROC) analysis was used to compare the ability of an antibody to discriminate between lung ADC and reactive pneumocyte hyperplasia.

EGFR mutations

12 (38)

0

p53 (threshold 30%)

13 (41)

3 (7.5)

p53 (threshold 50%)

1 (3.1)

MDM2

20 (63)

SFN EGFR MUTATION STATUS IN BIOPSY SAMPLES

To examine the utility of EGFR mutation-specific antibodies on small amounts of tissue obtained from biopsies, pre-operative biopsy samples were examined that corresponded to surgically resected cases testing positive for L858R or E746_A750 by IHC.

Results SURGICAL SPECIMENS

Of the 32 lung ADC cases evaluated, EGFR mutations were detected in 12 (38%) specimens: nine (28%) with the L858R antibody and three (9.4%) with the E746_A750 antibody. When expression patterns were classified as diffuse (>50%) or focal (10–50%), nine (75%) cases showed the former pattern, and three (25%) cases the latter. No samples tested positively with both antibodies. In contrast, all the reactive pneumocyte hyperplasia specimens were negative when treated with the EGFR mutation-specific antibodies. We did not observe immunopositivity with the EGFR mutation-specific antibodies during a wholesection analysis of normal lung and bronchial epithelium, although we detected immunoreactivity of smooth muscle cells around the normal bronchial and vascular components with clone 6B6. We examined the correlations between positive reactions obtained with EGFR mutation-specific IHC and molecular methods. Complete matches of EGFR status between IHC and molecular analysis in lung ADC cases were observed. Of the 32 ADCs, 13 (41%), one (3.1%) and 20 (63%) cases were positive for p53 (threshold 30%), p53 (threshold 50%) and MDM2, respectively, while none were positive for SFN. Of the 40 pneumocyte hyperplasias, p53 (threshold 30%), p53 (threshold 50%), MDM2 and SFN immunoreactivity was observed in three (7.5%), 0, 16 (40%) and three (7.5%) cases, respectively. These results are summa-

0

Benign diseases (n = 40) (%)

0 16 (40) 3 (7.5)

IHC, Immunohistochemistry; ADCs, adenocarcinomas; EGFR, epidermal growth factor receptor; MDM2, Mouse double minute 2; SFN, stratifin.

rized in Table 2, and representative images of tissue samples stained with each antibody are shown in Figure 1. ROC CURVE ANALYSIS

Analyses of ROC curves revealed that EGFR mutation-specific antibodies had the highest AUC value (0.688), followed by p53 threshold 30% (0.666), p53 threshold 50% (0.516), MDM2 (0.613) and SFN (0.463) (Table 3). The EGFR mutation-specific and p53 antibody pair showed the highest AUC value (0.744) among the combinations tested. EGFR STATUS IN BIOPSY SAMPLES

Of the 12 ADC cases that were identified as positive for EGFR mutations, pre-operative histological diagnoses from transbronchial biopsies revealed eight definite ADC cases, and four suspected cases for which the paucity in the number of tumorigenic or atypical cells precluded a definitive determination of malignancy. Of these 12 biopsies, EGFR mutations were detected in six ADC specimens and four cases in which atypical cells were present (Table 4 and Figure 2). The two biopsy samples that were negative for EGFR mutations by IHC corresponded to surgically resected specimens for which a focal staining pattern was observed using these antibodies.

Discussion The results of this study demonstrate a novel use for EGFR mutation-specific antibodies. Although their sensitivity for detecting lung ADC was suboptimal, these antibodies were able to discriminate between ADC and © 2014 John Wiley & Sons Ltd, Histopathology, 66, 816–823.

EGFR immunochemistry discriminating adenocarcinoma

(A)

(B)

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Table 3. The area under the curve value of each antibody and combinations of these antibodies IHC

AUC

EGFR mutations

0.688

p53 (threshold 30%)

0.666

p53 (threshold 50%)

0.516

MDM2

0.613

SFN

0.463

EGFR mutations and p53 (threshold 30%)

0.744

EGFR mutations and p53 (threshold 50%)

0.688

EGFR mutations and MDM2

0.659

EGFR mutations and SFN

0.650

p53 (threshold 30%) and MDM2

0.663

p53 (threshold 50%) and MDM2

0.613

p53 (threshold 30%) and SFN

0.628

p53 (threshold 50%) and SFN

0.478

MDM2 and SFN

0.588

IHC, Immunohistochemistry; AUC, area under the curve; EGFR, epidermal growth factor receptor; MDM2, Mouse double minute 2; SFN, stratifin. (C)

Figure 1. Representative specimens of benign pneumocyte hyperplasia obtained by surgical resection. A, Haematoxylin and eosin staining showing alveoli filled with granulation tissue and inflammatory cells. Expression of (B) p53 and (C) Mouse double minute 2 was detected by immunostaining.

benign pneumocyte hyperplasia. In addition, it was shown that detection using these antibodies could be applied effectively to small biopsy samples. © 2014 John Wiley & Sons Ltd, Histopathology, 66, 816–823.

The lepidic growth pattern was the focus of the current analysis, as it is observed in both non-invasive ADCs and reactive pneumocyte hyperplasias induced by inflammation. In contrast, a solid growth pattern is generally destructive, and can therefore be identified readily as a carcinoma. Cases are often difficult to distinguish as either lung ADC or benign disease.19 In addition, we reported previously that lepidic predominant tumours have the second-highest EGFR mutation rate among lung ADCs.20 The correlation between EGFR mutations and a lepidic growth pattern has been reported previously.21 In addition, ethnic differences in the frequency of EGFR mutations have been described,22 although one study of European cases harbouring a lepidic component found prevalence rates similar to those that we and others have found.23 Thus, EGFR mutational status can be used as a diagnostic criterion for proliferating atypical pneumocytes along with the presence of a pre-existing alveolar wall. The present data demonstrating that EGFR mutations are highly specific to ADCs are in accordance with a report that these mutations are never detected in normal lung tissue.23,24 By contrast, EGFR

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Table 4. EGFR-mutation IHC positive cases in surgical specimens and the results in bronchoscopic specimens Surgery specimens Case number

Histology

Bronchoscopy specimens

EGFR IHC

Type of EGFR antibody

EGFR PCR

Type of EGFR mutation

Diagnose

Type of EGFR antibody

EGFR IHC

4

Lepidic predominant

Positive

E746_A750

Positive

E746_A750

ADC

E746_A750

Positive

8

MIA

Positive

E746_A750

Positive

E746_A750

Atypical cells present

E746_A750

Positive

13

MIA

Positive

L858R

Positive

L858R

ADC

L858R

Positive

15

Lepidic predominant

Positive

L858R

Positive

L858R

ADC

L858R

Positive

16

MIA

Positive

L858R

Positive

L858R

ADC

L858R

Positive

19

Lepidic predominant

Positive

E746_A750

Positive

E746_A750

Atypical cells present

E746_A750

Positive

20

MIA

Positive

L858R

Positive

L858R

ADC

L858R

Negative

21

Lepidic predominant

Positive

L858R

Positive

L858R

ADC

L858R

Negative

23

AIS

Positive

L858R

Positive

L858R

ADC

L858R

Positive

24

AIS

Positive

L858R

Positive

L858R

Atypical cells present

L858R

Positive

25

Lepidic predominant

Positive

L858R

Positive

L858R

Atypical cells present

L858R

Positive

28

Lepidic predominant

Positive

L858R

Positive

L858R

ADC

L858R

Positive

EGFR, Epidermal growth factor receptor; IHC, immunohistochemistry; ADC, adenocarcinoma; PCR, polymerase chain reaction; MIA, minimally invasive adenocarcinoma; AIS, adenocarcinoma in situ.

mutations were present in 43% of morphologically normal bronchial and bronchiolar epithelial tissue samples from lung ADCs with EGFR mutations, but none of them from ADCs without mutations.25 A recent analysis also revealed that some morphologically normal bronchial epithelial samples from lung ADCs carrying EGFR mutations were positive for a novel L858R-specific antibody, which was not tested in the present study; however, 13 of the 16 cases (81%) were negative for this mutation according to sequencing.26 In addition, although the antibodies against L858R and E746_A750 have been shown to be highly specific, as we have mentioned in previous reports,12–15 they have some limitations; thus, they should be used meticulously, and a careful interpretation of the results is essential. With clone 6B6, in all cases we detected immunoreactivity of smooth muscle cells around the normal bronchial and vascular com-

ponents, which is a feature of this clone. A significant advantage of IHC using EGFR mutation-specific antibodies is that it enables the simultaneous visualization of mutated and atypical cells. ROC curve analyses revealed that EGFR mutationspecific antibodies had the highest AUC value, indicating excellent specificity – despite a suboptimal sensitivity – in lung ADCs, which was in agreement with previously reported EGFR mutation frequencies in ADCs of 10–50%.27–29 The ROC data showed that the use of antibodies against other ADC markers was less effective compared to L858R and E746_A750 for identifying lung ADCs. The combination of EGFR mutation-specific and p53 antibodies showed the highest AUC value. Kobel et al.30 mentioned that, for ovarian carcinoma, nuclear overexpression in more than 50% of tumour cells indicated TP53 mutation, and focal nuclear expression in up to 50% of tumour © 2014 John Wiley & Sons Ltd, Histopathology, 66, 816–823.

EGFR immunochemistry discriminating adenocarcinoma

(A)

(B)

(c)

(D)

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Figure 2. Representative lung adenocarcinoma specimens. A, A lepidic growth pattern was observed by haematoxylin and eosin staining. B, Expression of the mutant form of EGFR (L858R) was detected using a mutation-specific antibody. C, Atypical cells in a corresponding bronchoscopic biopsy specimen were observed by haematoxylin and eosin staining. D, EGFRL858R expression was detected in the bronchoscopic biopsy specimen. (A) and (B) from surgically resected specimen. (C) and (D) corresponding biopsy specimen.

cells indicated wild-type TP53. We use two thresholds, 30% and 50%; using a threshold of 30%, the sensitivity, specificity and AUC value were 41%, 93% and 0.666, respectively; and using a threshold of 50%, the sensitivity, specificity and AUC value were 3.1%, 100% and 0.516, respectively. Using a threshold defining ≥50% of tumour cells with nuclear staining as positive, the specificity increased to 100% but the sensitivity decreased to 3.1%. Nevertheless, as reported previously, p53 immunoreactivity was also observed in a subset of reactive pneumocyte hyperplasia cases. Benign diseases, such as chronic infections and inflammatory and interstitial lung diseases, are associated with an increased p53 mutation frequency.31–34 As such, caution should be exercised when using p53 detection as a diagnostic tool. EGFR mutation-specific antibodies were also useful in detecting a small number of atypical cells in biopsy © 2014 John Wiley & Sons Ltd, Histopathology, 66, 816–823.

specimens, which normally represents a diagnostic challenge. Sampling an adequate number of tumour cells is essential for biomarker analyses. Thus, in cases where few atypical cells are present in biopsy specimens with EGFR-mutation antibody-positive, the results would provide justification for subjecting the patient to a second biopsy or other invasive procedures. Furthermore, these antibodies can be used as a means of differentiating between primary lung ADC and metastatic cancers, similar to thyroid transcription factor-1.35 In summary, detecting specific EGFR mutations can be a clinically useful, adjunctive diagnostic tool for distinguishing between primary lung ADC and benign pneumocyte hyperplasia or metastatic cancer, or for assessing tumour response to EGFR kinase inhibitors,10 which will require further investigation in a larger number of cases.

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Conflict of interest The authors declare no conflicts of interest.

14.

Acknowledgements This work was supported by the National Cancer Center Research and Development Fund (23-A-2, 23A-11, 23-A-35, and 24-A-1), and Grant-in-Aid for Scientific Research (C; grant number 25460446). We thank Koh Furuta and Noriko Abe for their excellent technical assistance.

15.

16.

17.

References 1. Travis WD, Garg K, Franklin WA et al. Evolving concepts in the pathology and computed tomography imaging of lung adenocarcinoma and bronchioloalveolar carcinoma. J. Clin. Oncol. 2005; 23; 3279–3287. 2. Aoki T, Hanamiya M, Uramoto H et al. Adenocarcinomas with predominant ground-glass opacity: correlation of morphology and molecular biomarkers. Radiology 2012; 264; 590–596. 3. Collins J, Stern EJ. Ground-glass opacity at CT: the ABCs. Am. J. Roentgenol. 1997; 169; 355–367. 4. Aoki T, Tomoda Y, Watanabe H et al. Peripheral lung adenocarcinoma: correlation of thin-section CT findings with histologic prognostic factors and survival. Radiology 2001; 220; 803–809. 5. Brandman S, Ko JP. Pulmonary nodule detection, characterization, and management with multidetector computed tomography. J. Thorac. Imaging 2011; 26; 90–105. 6. Nomori H, Watanabe K, Ohtsuka T et al. Evaluation of F-18 fluorodeoxyglucose (FDG) PET scanning for pulmonary nodules less than 3 cm in diameter, with special reference to the CT images. Lung Cancer 2004; 45; 19–27. 7. Henschke CIYD, Mirtcheva R, McGuinness G, McCauley D, Miettinen OS, ELCAP Group. CT screening for lung cancer: frequency and significance of part-solid and nonsolid nodules. Am. J. Roentgenol. 2002; 178; 1053–1057. 8. Lee HY, Lee KS. Ground-glass opacity nodules histopathology, imaging evaluation, and clinical implications. J. Thorac. Imaging 2011; 26; 106–118. 9. Takeda Y, Tsuta K, Shibuki Y et al. Analysis of expression patterns of breast cancer-specific markers (mammaglobin and gross cystic disease fluid protein 15) in lung and pleural tumors. Arch. Pathol. Lab. Med. 2008; 132; 239–243. 10. Lynch TJ, Bell DW, Sordella R et al. Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N. Engl. J. Med. 2004; 350; 2129–2139. 11. Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat. Rev. Cancer 2007; 7; 169–181. 12. Kozu Y, Tsuta K, Kohno T et al. The usefulness of mutationspecific antibodies in detecting epidermal growth factor receptor mutations and in predicting response to tyrosine kinase inhibitor therapy in lung adenocarcinoma. Lung Cancer 2011; 73; 45–50. 13. Kato Y, Peled N, Wynes MW et al. Novel epidermal growth factor receptor mutation-specific antibodies for non-small cell lung

18.

19. 20.

21.

22.

23.

24.

25.

26.

27.

28.

29.

cancer: immunohistochemistry as a possible screening method for epidermal growth factor receptor mutations. J. Thorac. Oncol. 2010; 5; 1551–1558. Kawahara A, Yamamoto C, Nakashima K et al. Molecular diagnosis of activating EGFR mutations in non-small cell lung cancer using mutation-specific antibodies for immunohistochemical analysis. Clin. Cancer Res. 2010; 16; 3163–3170. Kawahara A, Taira T, Azuma K et al. A diagnostic algorithm using EGFR mutation-specific antibodies for rapid response EGFR-TKI treatment in patients with non-small cell lung cancer. Lung Cancer 2012; 78; 39–44. Xue Q, Sano T, Kashiwabara K et al. Aberrant expression of pRb, p16, p14ARF, MDM2, p21 and p53 in stage I adenocarcinomas of the lung. Pathol. Int. 2002; 52; 103–109. Shiba-Ishii A, Kano J, Morishita Y et al. High expression of stratifin is a universal abnormality during the course of malignant progression of early-stage lung adenocarcinoma. Int. J. Cancer 2011; 129; 2445–2453. 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; 6; 244–285. Mills SE ed. Histology for pathologists, 3rd edn. Philadelphia, PA: Lippincott Williams & Wilkins, 2007; 493–495. Tsuta K, Kawago M, Inoue E et al. The utility of the proposed IASLC/ATS/ERS lung adenocarcinoma subtypes for disease prognosis and correlation of driver gene alterations. Lung Cancer 2013; 81; 371–376. Yoshizawa A, Sumiyoshi S, Sonobe M et al. Validation of the IASLC/ATS/ERS lung adenocarcinoma classification for prognosis and association with EGFR and KRAS gene mutations: analysis of 440 Japanese patients. J. Thorac. Oncol. 2013; 8; 52–61. Mitsudomi T, Yatabe Y. Mutations of the epidermal growth factor receptor gene and related genes as determinants of epidermal growth factor receptor tyrosine kinase inhibitors sensitivity in lung cancer. Cancer Sci. 2007; 98; 1817–1824. Blons H, Cote JF, Le Corre D et al. Epidermal growth factor receptor mutation in lung cancer are linked to bronchioloalveolar differentiation. Am. J. Surg. Pathol. 2006; 30; 1309–1315. Shigematsu H, Lin L, Takahashi T et al. Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers. J. Natl Cancer Inst. 2005; 97; 339– 346. Tang X, Shigematsu H, Bekele BN et al. EGFR tyrosine kinase domain mutations are detected in histologically normal respiratory epithelium in lung cancer patients. Cancer Res. 2005; 65; 7568–7572. Allo G, Bandarchi B, Yanagawa N et al. Epidermal growth factor receptor mutation-specific immunohistochemical antibodies in lung adenocarcinoma. Histopathology 2014; 64; 826–839. Mitsudomi T, Kosaka T, Endoh H et al. Mutations of the epidermal growth factor receptor gene predict prolonged survival after gefitinib treatment in patients with non-small-cell lung cancer with postoperative recurrence. J. Clin. Oncol. 2005; 23; 2513–2520. Mitsudomi T, Kosaka T, Yatabe Y. Biological and clinical implications of EGFR mutations in lung cancer. Int. J. Clin. Oncol. 2006; 11; 190–198. Kosaka T, Yatabe Y, Endoh H et al. Mutations of the epidermal growth factor receptor gene in lung cancer: biological and clinical implications. Cancer Res. 2004; 64; 8919–8923. © 2014 John Wiley & Sons Ltd, Histopathology, 66, 816–823.

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30. Kobel M, Reuss A, Bois A et al. The biological and clinical value of p53 expression in pelvic high-grade serous carcinomas. J. Pathol. 2012; 222; 191–198. 31. Kunitake R, Kuwano K, Miyazaki H et al. Expression of p53, p21 (Waf1/Cip1/Sdi1) and Fas antigen in collagen vascular and granulomatous lung diseases. Eur. Respir. J. 1998; 12; 920–925. 32. Hojo S, Fujita J, Yamadori I et al. Overexpression of p53 protein in interstitial lung diseases. Respir. Med. 1998; 92; 184– 190. 33. Hofseth LJ, Saito S, Hussain SP et al. Nitric oxide-induced cellular stress and p53 activation in chronic inflammation. Proc. Natl Acad. Sci. USA 2003; 100; 143–148. 34. Staib F, Robles AI, Varticovski L et al. The p53 tumor suppressor network is a key responder to microenvironmental compo-

© 2014 John Wiley & Sons Ltd, Histopathology, 66, 816–823.

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nents of chronic inflammatory stress. Cancer Res. 2005; 65; 10255–10264. 35. Wen YH, Brogi E, Hasanovic A et al. Immunohistochemical staining with EGFR mutation-specific antibodies: high specificity as a diagnostic marker for lung adenocarcinoma. Mod. Pathol. 2013; 26; 1197–1203.

Supporting Information Additional Supporting Information may be found in the online version of this article: Table S1. Clinical and pathological characteristics of adenocarcinomas (n = 32).

Novel use for an EGFR mutation-specific antibody in discriminating lung adenocarcinoma from reactive pneumocyte hyperplasia.

Pulmonary ground-glass nodules (GGNs) are frequently observed. Histopathologically, their presentation can indicate a wide range of disorders from an ...
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