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(Chest. 2001;120:589-594.)
© 2001 American College of Chest Physicians

Association of p53 Gene Mutation and Telomerase Activity in Resectable Non-Small Cell Lung Cancer*

Yoshimasa Maniwa, MD, FCCP; Masahiro Yoshimura, MD; Chiho Obayashi, MD; Mayumi Inaba, MD; Kazue Kiyooka, MD; Makiko Kanki, MD and Yutaka Okita, MD

* From the Departments of Surgery Division II (Drs. Maniwa, Yoshimura, Kiyooka, Kanki, and Okita) and Pathology (Drs. Obayashi and Inaba), Kobe University School of Medicine, Kobe, Japan.

Correspondence to: Yoshimasa Maniwa, MD, FCCP, Department of Surgery Division II, Kobe University School of Medicine, 7–5-2 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan; e-mail: rk3y-mnw{at}asahi-net.or.jp


   Abstract
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 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Purpose: Mutation of the p53 gene and deregulation of telomerase may be essential for canceration in some malignant diseases. However, relationships between these occurrences have not yet been clarified. We examined the roles of p53 gene mutation and telomerase activity relative to the clinical and pathologic features of non-small cell lung carcinoma (NSCLC).

Methods: Frozen sections of 40 surgically resected NSCLC specimens were used. DNA extracted from fresh tumor specimens was analyzed with polymerase chain reaction (PCR), single-strand conformation polymorphism (SSCP) method, to screen alterations in the p53 gene. Exons showing aberrant band shifts on SSCP were reamplified, and the PCR products were directly sequenced. In addition, the telomerase activity of the same specimens was analyzed quantitatively with the fluorescence-based telomeric repeat amplification protocol assay, and the total product generated (TPG) method. Clinical and pathologic parameters were evaluated using a statistical analysis system.

Results: Mutations of the p53 gene relevant to an altered protein were confirmed in 19 of 40 specimens (47.5%). The TPG of 40 specimens was 75.24 ± 15.55 (mean ± SE). The TPG of the 19 specimens positive for p53 gene mutation was significantly higher than that of the 21 specimens negative for p53 gene mutation. Furthermore, the degree of cell differentiation was significantly correlated with both p53 gene mutation and high telomerase activity.

Conclusions: p53 gene mutation and high telomerase activity cooperate to induce tumorigenesis and low-grade differentiation in NSCLC. Simultaneous occurrence of p53 gene mutation and high telomerase activity may be relevant to the grade of malignancy in NSCLC.

Key Words: gene mutation • lung carcinoma • non-small cell lung cancer • p53 • telomerase • telomere


   Introduction
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 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Both mutation of the p53 gene and deregulation of telomerase are considered to be important events in the development and progression of cancer, and a relationship between p53 gene mutation and telomerase activity was suggested in previous basic studies.1 2 3 4 5 In the present study, p53 gene mutation and telomerase activity in cancer cells obtained from surgical specimens of non-small cell lung carcinoma (NSCLC) were assayed to investigate the relationship between p53 and telomerase. In addition, we examined this relationship in the context of various clinical and histopathologic parameters.


   Materials and Methods
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 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Forty specimens were sampled from 40 surgical cases of NSCLC between May 1997 and May 1999 (squamous cell carcinoma [n = 21], adenocarcinoma [n = 17], large cell carcinoma [n = 2]). These samples were histologically confirmed to be a part of tumor masses. Samples were frozen in liquid nitrogen immediately after surgical resection and stored at - 80°C until use.

Polymerase Chain Reaction, Single-Strand Conformation Polymorphism Method
To screen alternations of the p53 gene, the coding region encompassing exons 5 to 8, in which 90% of aberrations were concentrated,6 was analyzed by polymerase chain reaction (PCR), single-stand conformation polymorphism (SSCP) method. DNA was extracted according to a method described previously.7 Primer sets for amplification of four exons of p53 were designed with fluorescence Cy-5 (Amersham Pharmacia Biotech) at the 5' site of primers according to gene bank X54156, using the following primers:

exon 5: 5'-TTCCTCTTCCTACAGTACTCC-3'

and 5'-GCCCCAGCTGCTCACCATCGC-3',

exon 6: 5'-CACTGATTGCTCTTAGGTCTG-3'

and 5' -AGTTGCAAACCAGACCTCAGG-3',

exon 7: 5'-CCAAGGCGCACTGGCCTCATC-3'

and 5'-TCAGCGGCAAGCAGAGGCTGG-3',

exon 8: 5'-CCTATCCTGAGTAGTGGTAAT-3'

and 5'-GTCCTGCTTGCTTACCTCGCT-3'

PCR-SSCP analysis was performed according to Orita et al8 with the Expand High Fidelity PCR System (Roche Molecular Biochemicals; Mannheim, Germany) and by following PCR conditions of exons 5, 6, and 8 at 94°C for 30 s (denature), 60°C for 60 s (annealing), 72°C for 60 s (extension) for 35 cycles, exon 7 at 94°C for 30 s (denature), 70°C for 60 s (annealing), and 72°C for 60 s (extension) for 35 cycles. The PCR products were diluted 50-fold with 95% formamide and denatured at 80°C for 5 min, followed by rapid cooling on ice. Denatured products were separated on 5% polyacrylamide gels containing 5% glycerol with an automated laser fluorescence DNA sequencer with ALF Express (Pharmacia Biotech; Uppsala, Sweden) and analyzed with software (Allele Link; Pharmacia Biotech; Fig 1 , left, a).



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  		Figure 1. Sequence analysis of DNA prepared from a tumor and a normal control. Human placental DNA were used as the normal control. Left, a: PCR-SSCP analysis showing mobility shift in tumor DNA. Red arrows indicate the abnormal fragment in exon 6. Black lines indicate DNA from the healthy control, red lines and the DNA from the tumor. Right, b: Automated sequencing of exon 6. Missense mutation at codon 204 is shown. This G to T transition resulted in a Glu to Stop alteration.

 
DNA Sequencing
The exons showing aberrant band shifts on SSCP were reamplified from the original DNA samples with the Expand High Fidelity PCR System (exons 5 and 6 using primers 5'-TATCTGTTCACTTGTGCCCT-3' and 5'-GAGGGCCACTGACAACCACCC-3', exons 7 and 8 using primers 5'-CCTGCTTGCCACAGGTCTCCC-3' and 5'-CAAATGCCCCAATTGCAGGTA-3').

The PCR products were purified with a highly pure PCR product purification kit (Roche Molecular Biochemicals) and directly sequenced using a Thermosequenase kit (Amersham; Little Chalfont, UK) with ALF Express. The sequence was finally compared with the wild-type p53 sequence (Fig 1 , right, b).

Quantitative Analysis of Telomerase Activity
Telomerase activity of the same specimens was also analyzed quantitatively with the fluorescence-based telomeric repeat amplification protocol (TRAP) assay9 10 11 and the total product generated (TPG)12 method. The TRAP-eze Telomerase Detection Kit (Oncor; Gaithersburg, MD) was used according to the instructions of the manufacturer. Briefly, 5-mg frozen samples were homogenized in 100 L of ice-cold CHAPS lysis buffer (TRAP-eze) and then incubated for 30 min on ice. After incubation, the lysates were centrifuged at 12,000g for 20 min at 4°C. The supernatants were rapidly frozen and were stored at - 80°C. The concentration of protein was determined by Coomassie protein assay reagent (Pierce Chemical; Rockford, IL), and an aliquot of extract containing 1 µg of protein was used for each TRAP assay. Aliquots of extract were incubated with 0.1 ng Cy-5 labeled TS primer (5'-AATCCGTCGAGCAGAGTT-3') in a master mix (TRAP-eze). After 30-min incubation at 30°C, PCR was performed at 30 cycles (94°C for 30 s, 60°C for 30 s, 72°C for 45 s). The external control was a TSR8 (TRAP-eze) as a positive control. The products were applied (5 L/lane) to a 10% denaturing gel containing 6 mol/L urea, and fitted to an automated DNA sequencer. The first peak ladders obtained through the sequencer represented the Cy-5-labeled TS primer (18 base pair [bp]). The second peak represented the internal PCR control, yielding a 36-bp product (designated TSNT), which migrated in the analytic polyacrylamide gel at a position 14 bp below the smallest TRAP band. This control was used to monitor PCR efficiency during the PCR step of the assay. Lanes of samples the generated signals that formed a ladder with 6-bp increments from the third peak of 50 bp, represented the first amplifiable product. Findings from the sequencer were collected and analyzed automatically by Allele Links software (Pharmacia Biotech). Each peak was quantified in terms of height and area. The quantification of telomerase activity was determined by the formula: TPG (units/microgram protein) = [(T - B)/(CT)]/[(TSR8 - B)/(CTSR8)] x 100, where T = measured total area of telomerase activity (50 bp, 56 bp, 62 bp, 68 bp... ), B = measured area of the negative control reaction (background), TSR8 = measured total area of telomerase activity (50 bp, 56 bp, 62 bp, 68 bp... ) in external control, CT = measured area of internal control (36 bp), and CTSR8 = measured area of internal control (36 bp) in external control.

TNM Staging and Criteria for the Degree of Tumor Differentiation
For TNM staging of cancers, newly adopted (1997)13 American Joint Committee for Cancer criteria were used. The degree of histologic differentiation was determined by two pathologists using the parts of same specimen from which DNA was extracted. We divided adenocarcinoma and squamous cell carcinoma into three pathologic subclassifications according structural and cytologic atypia. Structural index of differentiation in adenocarcinoma was replacemental growth along alveolar walls, and tubular and papillary structure. Squamous cell carcinoma was evaluated by keratinization, intercellular bridges, and stratum with polarity. Most cases of poorly differentiated carcinomas, including adenocarcinoma and squamous cell carcinoma, revealed solid pattern.


   Results
TOP
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Mutations of the p53 gene were confirmed in 19 of 40 specimens (47.5%), 15 missense mutations, and 4 frameshift mutations. One case, in which the mutation did not cause the alteration of the amino acid, was excluded. The TPG of 40 specimens was 75.24 ± 15.55 (mean ± SE; Tables 1 and 2 ). TPG of the 19 specimens positive for p53 gene mutation was significantly higher than that of the 21 specimens negative for p53 gene mutation (Fig 2 , left, a). The examination of each histologic type showed the same finding (Fig 2 , right, b). No clinical or histopathologic parameters were related to p53 gene mutation or telomerase activity except cell differentiation (Fig 3 , 4 ). These tendencies were also observed in each histologic type.


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  		Table 1. Characteristics of 40 NSCLC Cases *

 


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  		Figure 2. Left, a: the TPG of 19 specimens positive for the p53 gene mutation was 119.45 ± 29.15 (mean ± SE), compared with 35.24 ± 6.13 of 21 specimens negative for the p53 gene mutation (p = 0.0005, Mann-Whitney U test). Right, b: in the cases of squamous cell carcinoma, the TPG of 9 specimens positive for the p53 gene mutation was 98.21 ± 25.03, compared with 47.61 ± 8.45 of 12 specimens negative for the p53 gene mutation (p = 0.047). In adenocarcinoma, TPG of eight specimens positive for p53 gene mutation was 98.81 ± 27.95, compared with 18.75 ± 5.32 of nine specimens negative for the p53 gene mutation (p = 0.009).

 


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  		Figure 3. The correlation of TPG with histologic differentiation. Left, a: the TPG of 15 specimens that were poorly differentiated was 98.00 ± 20.02 (mean ± SE), compared with 41.06 ± 7.52 of 23 well and moderately differentiated specimens (p = 0.003). Right, b: in the cases of squamous cell carcinoma, the TPG of 10 poorly differentiated specimens was 97.03 ± 23.52, compared with 44.08 ± 5.45 of 11 well and moderately differentiated specimens (p = 0.035). In adenocarcinoma, the TPG of 6 poorly differentiated specimens was 90.31 ± 35.17, compared with 37.95 ± 15.13 of 11 well and moderately differentiated specimens (p = 0.070).

 

   Discussion
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 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
The p53 tumor suppressor gene plays two important roles in genomic stability: blocking cell proliferation after DNA damage until it has been repaired,14 and starting apoptosis if the damage is too extensive.15 p53 mutations inactivate the tumor suppressor function of wild-type p53, elevating tumor incidences,16 and mutations are most frequently detected in human cancer.17 18 Activation of telomerase extends the telomere length and makes repeated cell divisions possible.9 19 20 Elevation of telomerase activity allows infinite cell divisions, resulting in immortalization of the cells. High telomerase activity was demonstrated in germ line cells and fetal cells as well as many tumor cells including lung carcinoma,21 22 and was applied to staging lymph nodes in patients with NSCLC.23 Mutation of p53 and deregulation of telomerase are considered to be the principal events in the development of cancer and the simultaneous occurrence of these events may be important for tumorigenesis in some cases. Even if p53 mutations are present, normal telomerase activity limits cell proliferation. Consequently, deregulation of telomerase activity may be necessary for tumorigenesis in p53 mutated cells.

The present clinical findings showed a relationship between telomerase activity and p53 mutation, providing a clue to the steps of tumorigenesis in NSCLC. Although the induction mechanism of abnormal telomerase activity has not yet been clarified, an association with the p53 gene24 and p53 protein overexpression25 has been proposed in lung cancer. Our previous study26 revealed the strong correlation between high telomerase activity and p53 protein overexpression. In contrast, some basic investigations27 28 concluded that expression of telomerase activity is observed in some immortal cell lines regardless of p53 status. Consequently, the initial disorder of telomerase activity in the development of cancer is possibly independent of p53 status. If the telomerase-activated cells are immortalized, their proliferation will be controlled by p53. However, repeated cell divisions assured by telomerase activation may increase the chance of p53 gene mutation. Once mutation of p53 is developed in such immortal cells, confused proliferation will start. Furthermore, p53 mutation enhances the telomerase activity through inactivation of p21waf1, which is induced by expression of wild-type p5329 30 and suppresses telomerase activity.31 32 The present findings exactly supported such reciprocal actions of deregulated telomerase activity and p53 mutation.

Grading of disease progression such as staging, T status, and N status did not directly reflect the biological characteristics of tumors because the duration from tumorgenesis to surgery varied widely among patients. The degree of histologic differentiation was more appropriate for presenting biological characteristics of the tumors although it was a qualitative assessment. In NSCLC, grading of histologic differentiation is well correlated with the proliferative activity defined using Ki-67 labeling,33 and poor differentiation represents poor prognosis in patients who have undergone surgical resection.34 35 36 The findings of the present study suggested that p53 gene mutation and high telomerase activity cooperate to cause a high frequency of cell division and poor differentiation of tumor tissues in NSCLC. Additional and simultaneous disorder of the p53 gene and telomerase activity may be relevant to the grade of malignancy in NSCLC.


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  		Table 2. TPG Data

 


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  		Figure 4. The influence of the p53 gene mutation and telomerase activity on histologic differentiation. In the p53 mutated group, the TPG of nine specimens that were poorly differentiated was 132.42 ± 25.83 (mean ± SE), compared with 60.33 ± 18.32 of eight well and moderately differentiated specimens (p = 0.032). In the p53 normal group, the TPG of 6 poorly differentiated specimens was 46.38 ± 17.73, compared to 30.78 ± 5.02 of 15 well and moderately differentiated specimens (p = 0.588).

 

   Footnotes
 
Abbreviations: bp = base pair; NSCLC = non-small cell lung carcinoma; PCR = polymerase chain reaction; SSCP = single-strand conformation polymorphism; TPG = total product generated; TRAP = telomeric repeat amplification protocol

Received for publication July 7, 2000. Accepted for publication February 14, 2001.


   References
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 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Gollahon, LS, Shay, JW (1996) Immortalization of human mammary epithelial cells transfected with mutant p53 (273his). Oncogene 12,715-725[ISI][Medline]
  2. Klingelhutz, AJ, Foster, SA, Mcdougall, JK (1996) Telomerase activation by the E6 gene product of human papillomavirus type 16. Nature 380,79-82[CrossRef][Medline]
  3. Yokoyama, Y, Takahashi, Y, Morishita, S, et al (1997) Introduction of p21Waf1/Cip1 gene into a carcinoma cell line of the uterine cervix with inactivated p53. Cancer Lett 116,233-239[CrossRef][ISI][Medline]
  4. Maxwell, SA, Capp, D, Acosta, SA (1997) Telomerase activity in immortalized endothelial cells undergoing p53-mediated apoptosis. Biochem Biophys Res Commun 241,642-645[CrossRef][ISI][Medline]
  5. Cao, Y, Gao, Q, Wazer, DE, et al (1997) Abrogation of wild-type p53-mediated transactivation is insufficient for mutant p53-induced immortalization of normal human mammary epithelial cells. Cancer Res 57,5584-5589[Abstract/Free Full Text]
  6. Kishimoto, Y, Mutakami, Y, Shiraishi, M, et al (1992) Aberrations of the p53 tumor suppressor gene in human non-small cell carcinomas of the lung. Cancer Res 52,4799-4804[Abstract/Free Full Text]
  7. Sato, T, Tanigami, A, Yamakawa, K, et al (1990) Allelotype of breast cancer: cumulative allele losses promote tumor progression in primary breast cancer. Cancer Res 50,7184-7189[Abstract/Free Full Text]
  8. Orita, M, Suzuki, Y, Sekiya, T, et al (1989) Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5,874-879[CrossRef][ISI][Medline]
  9. Kim, NW, Piatyszek, MA, Prowse, KR, et al (1994) Specific association of a telomerase activity with immortal cells and cancer. Science 266,2011-2015[Abstract/Free Full Text]
  10. Wright, WE, Shay, JW, Piatyazek, MA (1995) Modifications of a telomeric repeat amplification protocol (TRAP) result in increased reliability, linearity and sensitivity. Nucleic Acid Res 23,3794-3795[Free Full Text]
  11. Hisatomi, H, Nagano, K, Komatsu, H (1997) Quantification of telomerase activity in human liver tissue by fluorescence-based TRAP analysis. Hepatol Res 7,35-42
  12. Albanell, J, Lonardo, F, Rusch, V, et al (1997) High telomerase activity in primary lung cancers: association with increased cell proliferation rates and advanced pathologic stage. J Natl Cancer Inst 89,1609-1615[Abstract/Free Full Text]
  13. Mountain, CF (1997) Revisions in the international system for staging lung cancer. Chest 111,1710-1717[Abstract/Free Full Text]
  14. Kastan, MB, Onyekwere, O, Sidransky, D, et al (1991) Participation of p53 protein in the cellular response to DNA damage. Cancer Res 51,6304-6311[ISI][Medline]
  15. Clarke, AR, Purdie, CA, Harrison, DJ, et al (1993) Thymocyte apoptosis induced by p53-dependent and independent and independent pathway. Nature 352,849-852
  16. Paulovich, AG, Toczyski, DP, Hartwell, LH (1997) When checkpoints fall. Cell 88,315-321[CrossRef][ISI][Medline]
  17. Hollstein, M, Sidransky, D, Vogelstein, B, et al (1991) p53 mutation in human cancer. Science 262,1980-1981
  18. Kikuchi-Yanoshita, R, Konishi, M, Ito, S, et al (1992) Genetic changes of both p53 alleles associated with the conversion from colorectal adenoma in familial adenomatous polyposis and non-familial adenomatous polyposis patients. Cancer Res 52,3965-3971[Abstract/Free Full Text]
  19. Greider, CW, Blackburn, EH (1985) Identification of a specific telomere terminal transferase activity in tetrahymera extracts. Cell 43,405-413[CrossRef][ISI][Medline]
  20. Morin, GB (1989) The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell 59,521-529[CrossRef][ISI][Medline]
  21. Harley, CB, Villeponteau, B (1995) Telomeres and telomerase in aging and cancer. Curr Opin Genet Dev 5,249-255[CrossRef][Medline]
  22. Xinarianos, G, Scott, FM, Liloglou, T, et al (1999) Telomerase activity in non-small cell lung carcinomas correlates with smoking status. Int J Oncol 15,961-965[ISI][Medline]
  23. Ahrendt, SA, Yang, SC, Wu, L, et al (1997) Comparison of oncogene mutation and telomerase activity for the molecular staging of non-small cell lung cancer. Clin Cancer Res 3,1207-1214[Abstract]
  24. Hiyama, K, Ishioka, S, Shirotani, Y, et al (1995) Alteration in telomeric repeat length in lung cancer are associated with loss of heterozygosity in p53 and Rb. Oncogene 10,937-944[ISI][Medline]
  25. Wu, X, Kemp, B, Amos, CI, et al (1999) Association among telomerase activity, p53 protein overexpression, and genetic instability in lung cancer. Br J Cancer 80,453-457[CrossRef][ISI][Medline]
  26. Kiyooka, K, Maniwa, Y, Okada, M (1999) Analysis of mutant p53 and telomerase activity in non-small cell lung cancer. Ann Thorac Cardiovasc Surg 5,293-299[Medline]
  27. Sakitani, H, Tsujiuchi, T, Kobitsu, K, et al (1998) Increased telomerase activity and absence of p53 mutation in oligoastrocytomas induced by N-etyhyl-N-nitrosoures in rats. Cancer Lett 126,157-164[CrossRef][ISI][Medline]
  28. Milas, M, Yu, D, Sun, D, et al (1998) Telomerase activity of sarcoma cell lines and fibroblasts is independent of p53 status. Clin Cancer Res 4,1573-1579[Abstract]
  29. el-Deiry, WS, Tokino, T, Velculescu, VE, et al (1993) WAF1, a potential mediator of p53 tumor suppression. Cell 75,817-825[CrossRef][ISI][Medline]
  30. Deng, C, Zhang, P, Harper, JW, et al (1995) Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 82,675-684[CrossRef][ISI][Medline]
  31. Kallassy, M, Martel, N, Damour, O, et al (1998) Growth arrest of immortalized human keratinocytes and suppression of telomerase activity by p21WAF1 gene expression. Mol Carcinog 21,26-36[CrossRef][ISI][Medline]
  32. Terasaki, M, Kato, S, Sugita, Y, et al (1999) Adenovirus-mediated over-expression of p21Waf-1 differentially affects the proliferation, telomerase activity, and expression of glial fibrillary acidic protein in human glioma cells. Int J Oncol 14,63-69[ISI][Medline]
  33. Kawai, T, Suzuki, M, Kono, S, et al (1994) Proliferating cell nuclear antigen and Ki-67 in lung carcinoma. Cancer 74,2468-2475[CrossRef][ISI][Medline]
  34. Harpole, DH, Jr, Herndon, JE, II, Wolfe, WG, et al (1995) Prognostic model of recurrence and death in stage I non-small cell lung cancer utilizing presentation, histopathology, and oncoprotein expression. Cancer Res 55,51-56[Abstract/Free Full Text]
  35. Chung, CK, Zaino, R, Stryker, JA, et al (1982) Carcinoma of the lung: evaluation of histologic grade and factors influencing prognosis. Ann Thorac Surg 33,599-604[Abstract]
  36. Ichinose, Y, Yano, T, Asoh, H, et al (1995) Prognostic factors obtained by a pathologic examination in completely resected non-small cell lung cancer. Thorac Cardiovasc Surg 110,601-605



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