Respiration 2005;72:249–253
(DOI:10.1159/000085365)

Surfactant Protein A Detection in Primary Pulmonary Adenocarcinoma without Bronchioloalveolar Pattern

Uzaslan E.a · Stuempel T.a · Ebsen M.a · Freudenberg N.b · Nakamura S.d · Costabel U.c · Guzman J.a
aGeneral and Experimental Pathology, University of Bochum, Bochum, bDepartment of Cytology, Institute of Pathology, University of Freiburg i. Br., Freiburg i. Br., cDepartment of Pneumology and Allergy, Ruhrlandklinik, Essen, Germany; dDepartment of General Medicine and Clinical Investigation, Nara Medical University, Nara, Japan
email Corresponding Author


 Outline


 goto top of outline Key Words

  • Adenocarcinoma of the lung
  • Surfactant protein A
  • Immunohistochemistry

 goto top of outline Abstract

Background: Immunohistochemical studies in human lung carcinoma reported positive staining of tumor cells for surfactant protein A (SP-A), especially in peripheral airway cell carcinoma, which include bronchioloalveolar carcinoma and in some reports also papillary subtypes. Objective: The purpose of this study was to determine the SP-A expression in tumor cells of lung adenocarcinoma without a bronchioloalveolar pattern, classified according to the WHO. Methods: In total, 169 primary adenocarcinomas of the lung (109 acinar, 32 solid with mucin, 24 papillary and 4 mucinous) were examined by immunohistochemistry for SP-A expression. Results: Twenty-five percent of acinar, 38% of papillary and 3% of solid adenocarcinoma with mucin showed a positive intracytoplasmic SP-A reaction of the tumor cells. None of the mucinous adenocarcinomas stained for SP-A. This study included the largest number of acinar adenocarcinomas and solid adenocarcinomas with mucin studied for SP-A. We clearly demonstrated that also primary lung adenocarcinoma without a bronchioloalveolar pattern can express SP-A. A positive staining of hyperplastic type II cells surrounding the tumors or entrapped in the tumor could clearly be differentiated from the SP-A-positive stain of tumor cells. Conclusion: These results support the theory that SP-A-producing cells may generate not only bronchioloalveolar and papillary carcinoma, but also other subtypes of lung adenocarcinoma.

Copyright © 2005 S. Karger AG, Basel


goto top of outline Introduction

Immunohistochemical studies performed in human lung carcinoma reported a positive staining of tumor cells for surfactant protein A (SP-A) in part of adenocarcinoma of the lung [1, 2, 3, 4], especially in peripheral airway cell carcinoma, which include bronchioloalveolar carcinoma and in some reports also papillary subtypes [5, 6, 7, 8, 9, 10, 11, 12]. Ueno et al. [2] indicated that more than 80% of bronchioloalveolar carcinoma and 50–60% of grade I–III invasive adenocarcinoma of the lung stain positive with SP-A, but they did not further classify their adenocarcinoma according to histological subtypes. Mizutani et al. [7] concluded that only tumors with alveolar type II and/or Clara cell type are SP-A positive.

However, Hirata et al. [13] indicated in a study of 33 cases of bronchial gland cell type of adenocarcinoma that 68% of these cases were SP-A positive. Ten Have-Opbroek et al. [8] showed in a canine bronchogenic carcinoma model that alveolar type II tumor stem cells may generate not only adenocarcinoma with bronchioloalveolar and papillary patterns but also adenocarcinoma with other growth patterns. SP-A mRNA expression was also described in solid and glandular lung adenocarcinoma [14]. Different percentages of SP-A-positive tumors were found in other subtypes of adenocarcinoma (without bronchioloalveolar or papillary patterns), without further subclassifications of the tumor [9, 15], or in mixed acinar/papillary [11] or acinar/solid groups [5]. Only Sugiyama et al. [16] clearly classified 54 primary lung adenocarcinomas according to the WHO classification and found SP-A expression in tumors without a bronchioloalveolar pattern.

Therefore, the purpose of this study was to determine the SP-A expression in a large group of lung adenocarcinomas without bronchioloalveolar patterns, classified according to the WHO [17], because the presence of SP-A in these tumors may indicate that they also originate from type II or Clara cells.

 

goto top of outline Materials and Methods

goto top of outline Study Population

All primary adenocarcinomas of the lung without bronchioloalveolar patterns surgically resected by either lobectomy or pneumonectomy in the Ruhrlandklinik in the years 1992–1997 were included in this study. All cases were reclassified by three pathologists in consensus according to the latest edition of the WHO histological typing of lung tumors [17]. Bronchioloalveolar adenocarcinoma and mixed subtypes with a bronchioloalveolar pattern were excluded from this analysis.

In total, 169 primary adenocarcinomas of the lung, namely 109 acinar (ICD-0-8550/3), 32 solid with mucin (ICD-0-8230/3), 24 papillary (ICD-0-8260/3) and 4 mucinous ‘colloid’ adenocarcinomas (ICD-0-8480/3), were available.

goto top of outline Immunohistochemical Analysis

The cases were examined by immunohistochemistry using a modified peroxidase-antiperoxidase (PAP) technique as described before [18]. A polyclonal antibody against pulmonary SP-A was used with diaminobenzidine as chromogen (a gift from W. Bartsch, Fraunhofer Institute, Hannover, Germany) [19]. The quality of this antibody against SP-A was found to be comparable with commercially available antibodies. Positive and negative control sections for SP-A and for the specificity of the immunoreaction were included in each reaction. We defined as a diffusely positive immunostain when most of the tumor cells were positive at different fields in a 100× magnification and as a focally positive immunostain when few positive tumor cell groups were found disseminated or focally located in tumor tissue at different fields in a 100× magnification. The stained slides were read microscopically by three pathologists in consensus.

 

goto top of outline Results

Twenty-five percent of acinar adenocarcinoma, 38% of papillary adenocarcinoma and 3% of solid adenocarcinoma with mucin production (fig. 1, 2a, b, 3a, b) showed a positive intracytoplasmic SP-A reaction of tumor cells. The staining patterns of the tumors were diffusely positive in 25 cases (17 acinar, 8 papillary) and focally positive in 12 cases (10 acinar, 1 papillary, 1 solid with mucin). None of the mucinous adenocarcinomas stained for SP-A (fig. 4).

FIG01

Fig. 1. SP-A reaction in different subtypes of adenocarcinoma in our study.

FIG02

Fig. 2.a Acinar adenocarcinoma of the lung. Hematoxylin-eosin stain. Original magnification ×50. b Acinar adenocarcinoma with positive SP-A staining. PAP method. Original magnification ×50.

FIG03

Fig. 3.a Solid adenocarcinoma with mucin. PAS stain. Original magnification ×100. b Solid adenocarcinoma with mucin. Positive SP-A staining. PAP method. Original magnification ×100.

FIG04

Fig. 4.a Papillary adenocarcinoma of the lung. Hematoxylin-eosin stain. Original magnification ×50. b Papillary adenocarcinoma with positive SP-A staining. PAP method. Original magnification ×50.

The positive SP-A staining of hyperplastic type II cells surrounding the tumors or in some cases entrapped between the tumor infiltrates did not disturb the recognition of SP-A-positive tumor cells.

 

goto top of outline Discussion

In this study, we clearly demonstrate that also primary lung adenocarcinoma without a bronchioloalveolar pattern can express SP-A, which confirms the recently published observation by Ueno et al. [2]. However, these authors did not perform further subclassifications of the histological subtypes, as was done in our study. We studied the largest number of acinar adenocarcinomas and solid adenocarcinomas with mucin for SP-A expression described in the literature. We found that 25% of acinar adenocarcinoma and 3% of solid adenocarcinoma with mucin showed intracytoplasmic SP-A staining of the tumor cells. In our study, the percentage of SP-A-positive cases of adenocarcinoma without bronchioloalveolar patterns is lower than in the reports by Ueno et al. [2], by Sugiyama et al. [16] and by Hirata et al. [13], possibly because we excluded adenocarcinoma with mixed subtypes showing bronchioloalveolar or papillary components from this study, and because we very carefully discriminated the SP-A-positive staining of hyperplastic type II cells surrounding the tumors or entrapped in the tumor from the intracytoplasmic positive SP-A stain of tumor cells [20], which sometimes can be difficult.

We also studied true papillary lung adenocarcinoma diagnosed according to the WHO classification and other reports [21]. Thirty-eight percent of the true papillary carcinoma of our study were SP-A positive.

In contrast to Mizutani et al. [7], who concluded that only tumors with alveolar type II and/or Clara cell type are SP-A positive, our results support the theory [13, 22] that SP-A-producing cells may generate not only bronchioloalveolar and papillary carcinoma, but also other subtypes of lung adenocarcinoma such as lung acinar adenocarcinoma and solid adenocarcinoma with mucin.

 

goto top of outline Acknowledgement

This study was supported by a grant of the Ichiro Kanehara Foundation, Chihiro and Yokochi Fund.

Dr. Esra Uzaslan contributed to this study as a guest researcher at General and Experimental Pathology, University of Bochum, originally from Chest Diseases Department, Medical Faculty, University of Uludag, Bursa, Turkey.


 goto top of outline References
  1. Zamecnik J, Kodet R: Value of thyroid transcription factor-1 and surfactant apoprotein A in the differential diagnosis of pulmonary carcinomas: A study of 109 cases. Virchows Arch 2002;440:353–361.
  2. Ueno T, Linder S, Elmberger G: Aspartic proteinase napsin is a useful marker for diagnosis of primary lung adenocarcinoma. Br J Cancer 2003;88:1229–1233.
  3. Singh G, Scheithauer BW, Katyal SL: The pathobiologic features of carcinomas of type II pneumocytes. An immunocytologic study. Cancer 1986;57:994–999.
  4. Chieng DC, Cangiarella JF, Zakowski MF, Goswami S, Cohen JM, Yee HT: Use of thyroid transcription factor 1, PE-10, and cytokeratins 7 and 20 in discriminating between primary lung carcinomas and metastatic lesions in fine-needle aspiration biopsy specimens. Cancer 2001;93:330–336.
  5. Linnoila RI, Jensen SM, Steinberg SM, Mulshine JL, Eggleston JC, Gazdar AF: Peripheral airway cell marker expression in non-small cell lung carcinoma. Association with distinct clinicopathologic features. Am J Clin Pathol 1992;97:233–243.
  6. Komatsu H, Katayama T, Murakami K, Sagara Y, Hayashi K: Immunohistochemical study of lung cancer using monoclonal antibody against human pulmonary surfactant apoprotein. Nippon Kyobu Geka Gakkai Zasshi 1991;39:2172–2175.
  7. Mizutani Y, Nakajima T, Morinaga S, Gotoh M, Shimosato Y, Akino T, Suzuki A: Immunohistochemical localization of pulmonary surfactant apoproteins in various lung tumors. Special reference to nonmucus producing lung adenocarcinomas. Cancer 1988;61:532–537.
  8. Ten Have-Opbroek AA, Benfield JR, van Krieken JH, Dijkman JH: The alveolar type II cell is a pluripotential stem cell in the genesis of human adenocarcinomas and squamous cell carcinomas. Histol Histopathol 1997;12:319–336.
  9. Linnoila RI, Mulshine JL, Steinberg SM, Gazdar AF: Expression of surfactant-associated protein in non-small-cell lung cancer: A discriminant between biologic subsets. J Natl Cancer Inst Monogr 1992;13:61–66.

    External Resources

  10. Kitinya JN, Sueishi K, Tanaka K, Katsuda Y: Immunoreactivity of surfactant-apoprotein in adenocarcinomas, large cell and small cell carcinomas of the lung. Acta Pathol Jpn 1986;36:1271–1278.
  11. Kaufmann O, Dietel M: Thyroid transcription factor-1 is the superior immunohistochemical marker for pulmonary adenocarcinomas and large cell carcinomas compared to surfactant proteins A and B. Histopathology 2000;36:8–16.
  12. Gazdar AF, Linnoila RI, Kurita Y, Oie HK, Mulshine JL, Clark JC, Whitsett JA: Peripheral airway cell differentiation in human lung cancer cell lines. Cancer Res 1990;50:5481–5487.
  13. Hirata H, Noguchi M, Shimosato Y, Uei Y, Goya T: Clinicopathologic and immunohistochemical characteristics of bronchial gland type adenocarcinoma of the lung. Am J Clin Pathol 1990;93:20–25.
  14. Broers JL, Jensen SM, Travis WD, Pass H, Whitsett JA, Singh G, Katyal SL, Gazdar AF, Minna JD, Linnoila RI: Expression of surfactant associated protein-A and Clara cell 10 kilodalton mRNA in neoplastic and non-neoplastic human lung tissue as detected by in situ hybridization. Lab Invest 1992;66:337–346.
  15. Nicholson AG, McCormick CJ, Shimosato Y, Butcher DN, Sheppard MN: The value of PE-10, a monoclonal antibody against pulmonary surfactant, in distinguishing primary and metastatic lung tumours. Histopathology 1995;27:57–60.
  16. Sugiyama K, Kawai T, Nakanishi K, Suzuki M: Histochemical reactivities of lectins and surfactant apoprotein in pulmonary adenocarcinomas and their metastases. Mod Pathol 1992;5:273–276.
  17. Travis WD, Colby TV, Corrin B, Shimosato Y, Brambilla E: World Health Organisation (WHO) International Histologic Classification of Tumors. Histological Typing of Lung and Pleural Tumors, ed 3. WHO, 1999, pp 54–55.
  18. Bross KJ, Pangalis GA, Staatz CG, Blume KG: Demonstration of cell surface antigens and their antibodies by the peroxidase-antiperoxidase method. Transplantation 1978;25:331–334.
  19. Guzman J, Wang YM, Kalaycioglu O, Schoenfeld B, Hamm H, Bartsch H, Costabel U: Increased surfactant protein A content in human alveolar macrophages in hypersensitivity pneumonitis. Acta Cytol 1992;36:668–673.
  20. Singh G, Katyal SL, Ordonez NG, Dail DH, Negishi Y, Weedn VW, Marcus PB, Weldon-Linne CM, Axiotis CA, Alvarez-Fernandez E, Smith WI: Type II pneumocytes in pulmonary tumors. Implications for histogenesis. Arch Pathol Lab Med 1984;108:44–48.
  21. Silver SA, Askin FB: True papillary carcinoma of the lung: A distinct clinicopathologic entity. Am J Surg Pathol 1997;1:43–51.
  22. Ten Have-Opbroek AA, Hammond WG, Benfield JR, Teplitz RL, Dijkman JH: Expression of alveolar type II cell markers in acinar adenocarcinomas and adenoid cystic carcinomas arising from segmental bronchi. A study in heterotopic bronchogenic carcinoma model in dogs. Am J Pathol 1993;142:1251–1264.

 goto top of outline Author Contacts

Prof. Dr. med. J. Guzman y Rotaeche
c/o Prof. Shinobu Nakamura, Department of General Medicine and Clinical Investigation
Nara Medical University, 840 Shijo
Kashihara, Nara 634-8522 (Japan)
Tel. +81 744 29 8905, Fax +81 744 24 5739, E-Mail josune@naramed-u.ac.jp


 goto top of outline Article Information

Received: September 3, 2003
Accepted after revision: August 25, 2004
Number of Print Pages : 5
Number of Figures : 4, Number of Tables : 0, Number of References : 22


 goto top of outline Publication Details

Respiration (International Journal of Thoracic Medicine)

Vol. 72, No. 3, Year 2005 (Cover Date: May-June 2005)

Journal Editor: C.T. Bolliger, Cape Town
ISSN: 0025–7931 (print), 1423–0356 (Online)

For additional information: http://www.karger.com/res


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