Clinical Cancer Research CTRC-AACR San Antonio Breast Cancer Symposium Infection and Cancer: Biology, Therapeutics, and Prevention
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Cell Growth & Differentiation
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tomizawa, Y.
Right arrow Articles by Mori, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tomizawa, Y.
Right arrow Articles by Mori, M.
Clinical Cancer Research Vol. 11, 6816-6822, October 1, 2005
© 2005 American Association for Cancer Research

Human Cancer Biology

Clinicopathologic Significance of the Mutations of the Epidermal Growth Factor Receptor Gene in Patients with Non–Small Cell Lung Cancer

Yoshio Tomizawa1, Hironobu Iijima1, Noriaki Sunaga1, Koji Sato1, Atsushi Takise1, Yoshimi Otani2, Shigefumi Tanaka3, Tatsuo Suga4, Ryusei Saito6, Tamotsu Ishizuka1, Kunio Dobashi1, John D. Minna7, Takashi Nakajima5 and Masatomo Mori1

Authors' Affiliations: Departments of 1 Medicine and Molecular Science, 2 Thoracic and Visceral Organ Surgery, 3 General Surgical Science, 4 Medicine and Biological Science, and 5 Tumor Pathology, Gunma University Graduate School of Medicine, Showa-machi, Maebashi, Gunma, Japan; 6 National Nishigunma Hospital, Kanai, Shibukawa, Gunma, Japan; and 7 Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas

Requests for reprints: Yoshio Tomizawa, Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, 3-39-15 Showa-machi, Maebashi, Gunma 371-8511, Japan. Phone: 27-220-7111 ext. 8123; Fax: 27-220-8136; E-mail: ytomizaw{at}showa.gunma-u.ac.jp.


   Abstract
Top
 Abstract
Materials and Methods
 Results
 Discussion
 References
 
Purpose: It has been reported that the mutations of epidermal growth factor receptor (EGFR) are detected in lung cancers. Studies of EGFR mutations in large numbers of patients' tumors with clinical data including response to EGFR tyrosine kinase directed therapy are needed to develop a robust database for clinical use. The purpose of the present study is to gain further insights into the significance of EGFR mutation in non–small cell lung cancer (NSCLC).

Experimental Design: We investigated the clinicopathologic significance of tyrosine kinase domain (exons 18-21) EGFR mutations in 120 patients with primary NSCLC and the correlation between EGFR mutation and sensitivity to gefitinib in an additional 20 NSCLC patients treated with gefitinib. In addition, onocogenic KRAS mutations and RASSF1A promoter methylation were determined in the same samples.

Results: EGFR mutation was detected in 29 of 120 (24%) tumors. All of the 29 (40%) mutations occurred in 72 adenocarcinomas. EGFR mutation was significantly more frequent in females (47%) than males (12%, P < 0.0001), in younger patients (38%) than older patients (10%, P = 0.0005), in nonsmokers (47%) than smokers (13%, P < 0.0001), and in well-differentiated tumors (39%) than moderately and poorly differentiated tumors (7%, P < 0.0001). Mutation of the EGFR gene was preferentially observed in advanced disease. Furthermore, EGFR mutations were detected in 11 of 14 (79%) responders, whereas none of six (0%) nonresponders had the mutation (P = 0.0022).

Conclusions: These results in Japanese (East Asian) patients indicated that EGFR mutation plays an important role in pathogenesis of lung adenocarcinoma.

Epidermal growth factor receptor (EGFR) is a 170-kDa tyrosine kinase that is expressed in most human tissues and is highly expressed in many human cancers including lung cancer (1–3). EGFR signaling is triggered by the binding of growth factors, such as EGF, resulting in dimerization of EGFR molecules or heterodimerization with other closely related receptors, such as HER-2/neu (4). Activation of the downstream pathways of EGFR leads to cell proliferation, differentiation, migration, motility, adhesion, protection from apoptosis, enhanced survival, and gene transcription (5, 6). Recently, targeting the critical genetic lesion associated with the proliferative signals has become an effective treatment in cancer therapies (7–9). Gefitinib is an orally administered small molecule that inhibits EGFR tyrosine phosphorylation (10). It is the first targeted agent to be approved for the treatment of the patients with advanced non–small cell lung cancer (NSCLC). The objective tumor response rates were 18.4% in IDEAL 1 and 11.8% in IDEAL 2 (11, 12). In the IDEAL 1 subgroup analysis, the response rate in Japanese patients (27.5%) was higher that in Europe, Australian, and South Africa patients (10.4%). Furthermore, these studies showed that women patients with adenocarcinoma histology had a higher chance to respond to gefitinib (11, 12).

Recently, it was reported that mutations of the EGFR gene have been detected in NSCLC (13–19), and these mutations were found in the tyrosine kinase domain in EGFR. These mutations enhance tyrosine kinase activity in response to EGF and are associated with sensitivity to gefitinib (13–15, 20), although tumor with the mutation T970M was resistant to gefitinib (21). These mutations were more frequent in women than in men, in adenocarcinoma than in other histologies, in nonsmokers than in smokers, and in patients from Japan than in patients from the United State (13–16, 18, 19). On the other hand, Huang et al. (17) reported there was no correlation of the EGFR mutation rate with gender and smoking history in Taiwanese patients. Therefore, the correlation between EGFR mutation and clinicopathologic features in NSCLCs needs studies of large numbers of patients.

In the present study, we examined the correlation of EGFR mutations with clinical and pathologic features in the 120 Japanese patients with NSCLC, and we analyzed the association between EGFR mutations and sensitivity to gefitinib in an additional 20 patients with NSCLC treated with gefitinib. We also analyzed the association of EGFR mutations with KRAS mutations and RASSF1A methylation, because EGFR plays an important role in Ras signal transduction.


   Materials and Methods
Top
 Abstract
 Materials and Methods
Results
 Discussion
 References
 
Tissue and patients. Primary lung tumors were obtained from 120 consecutive Japanese cases with NSCLC without gefitinib treatment. All patients underwent operations at the Gunma University Hospital or National Nishigunma Hospital, Gunma, between August 1991 and March 2001. None of the patients had been treated before surgery. Institutional review board permission and informed consent were obtained for all cases. Clinical information, such as gender, age at diagnosis, and smoking history, were obtained by review of medical records of the patients. The median age of the patients in the present study was 67 years. All available histology slides from each case were reviewed, and the diagnosis of histology and the grade of differentiation were confirmed using the WHO histologic classification (22). The differentiation was determined as follows. In adenocarcinoma, well-differentiated tumors were composed of the bronchioloalveolar or papillary pattern, moderately differentiated tumors were composed of the papillary or acinar pattern, and poorly differentiated tumor were composed of the solid pattern. In squamous cell carcinoma, the grade of the differentiation was determined by the degree of the keratinization. Detailed information on these patients is summarized in Table 1. The stage of the disease was postoperative pathologic stage. Therefore, the stage III and IV diseases included preoperative clinical stage I or II diseases. The stage IV diseases included cases who received biopsies by open lung or video-assisted thoracoscopic surgery for diagnosis. The tumors were frozen and stored at –80°C until DNA extraction. Genomic DNA was prepared by a previously described method (23) or by a QIAamp DNA Mini Kit (Qiagen, Tokyo, Japan).


View this table:
[in this window]
[in a new window]
  		Table 1. Characteristicts of 120 patients with NSCLC

 
To analyze the correlation between EGFR mutation and sensitivity of tumors to gefitinib, we reviewed the medical records to find out the patients with NSCLC treated with gefitinib. We found 82 patients treated with gefitinib. Nine tumors diagnosed by cytology and 33 tumor tissues that did not include enough cells were excluded from this analysis. Then, to avoid the false negative due to the tumor heterogeneity, 20 samples obtained by transbronchial lung biopsy or percutaneous lung biopsy in nonresponder were also excluded from this analysis. Finally, 20 patients with NSCLC treated with gefitinib were included of this analysis. Serial 5-µm sections were cut from archival, formalin-fixed, paraffin-embedded tissue of these 20 patients. All slides were stained with H&E, and one of the slides was coverslipped. The coverslipped slide was used as a guide to localize regions of interest for microdissection of the other slides. Microdissection from archival paraffin-embedded tissues was done using a Leica laser microdissection system (Wetzlar, Germany) from multiple microslides of each sample. DNA was prepared by previously described method (24).

Mutational analyses of the EGFR and KRAS genes. Four exons (exons 18-21) which code for tyrosine kinase domain of the EGFR gene and codons 12 and 13 of the KRAS gene were amplified by PCR with EGFR- and KRAS-specific oligonucleotide primers as shown in Table 2. For PCR reactions, after initial denaturation at 95°C for 15 minutes, 35 cycles each consisting of denaturation at 94°C for 30 seconds, annealing at 55°C or 65°C for 30 seconds, and strand elongation at 72°C for 1 minute were done to amplify DNA fragments followed by a final elongation 72°C for 5 minutes. PCR products from the fresh frozen samples were subjected to single-strand conformation polymorphism analysis (25). PCR products that detected sifted bands on single-strand conformational polymorphism analysis were sequenced with ABI 310 Genetic Analyzer (Applied Biosystems, Foster City, CA). PCR products from the microdissected samples were directly sequenced.


View this table:
[in this window]
[in a new window]
  		Table 2. Primer sequences used for mutational analyses of the EGFR and KRAS genes

 
Methylation analysis of the RASSF1A gene. We had previously examined the same 120 patients for RASSF1A methylation (26). Briefly, RASSF1A methylation was analyzed by methylation-specific PCR method using genomic DNAs treated by sodium bisulfite.

Statistical analysis. Fisher's exact test was used to examine the association of two categorical variables. P < 0.05 was considered statistically significant.


   Results
Top
 Abstract
 Materials and Methods
 Results
Discussion
 References
 
EGFR mutations in tumors from patients without gefitinib treatment. One hundred twenty NSCLCs were examined for mutations in exons 18 to 21 of the EGFR gene by PCR/single-strand conformational polymorphism analysis and direct DNA sequencing. Representative results of the PCR/single-strand conformational polymorphism analysis are shown in Fig. 1. Mutation of the EGFR genes was detected in 29 of 120 (24%) tumors. Of 29 mutations, the distribution of the mutations was that one was in exon 18, 16 were in exon 19, and 12 were in exon 21. No mutation was detected in exon 20. There was no tumor that had multiple mutations in a tumor. In exon 19, all of 16 mutations were small in-frame deletion of the kinase domain. Of 16 mutations, 10 were delE746-A750, two were delE746-T751insA, two were delL747-T751insP, one was E747-E749, and one was R748-S752insT. In exon 21, all of 12 mutations were from T to G at the second nucleotide of codon 858 resulting in substitution of leucine with arginine residue. The mutation in exon 18 was that the substitution of G to A at nucleotide 2156 results in amino acid substitution of G for D at codon 719.



View larger version (69K):
[in this window]
[in a new window]
  		Fig. 1. Representative results of PCR-SSCP analyses. Shifted bands are indicated by arrows.

 
Correlation between EGFR mutations and clinicopathologic features in patients with non–small cell lung cancer. We analyzed the relationship between mutation of the EGFR genes and the clinicopathologic characteristics of these patients. The results are summarized in Table 3. EGFR mutation was more frequent in females than males, in younger patients than older patients, and in nonsmokers than smokers. Mutation of the EGFR gene was preferentially observed in advanced-stage NSCLC. All of the mutations occurred in adenocarcinoma. The frequency of EGFR mutation in adenocarcinoma was 29 of 72 (40%). It is significantly higher than that in nonadenocarcinoma. EGFR mutation was more frequent in higher differentiated tumors. There was no correlation between EGFR mutation and vascular invasion, lymphatic permeation, and pleural involvement.


View this table:
[in this window]
[in a new window]
  		Table 3. Correlation of EGFR mutation with clinicopathologic characteristics of patients with NSCLC

 
Correlation between EGFR mutations and gefitinib responsiveness in patients with gefitinib treatment. To investigate whether mutations of the EGFR gene are associated with sensitivity to genitinib, we examined the EGFR mutations in 20 samples obtained from patients with NSCLC treated with gefitinib. All patients were selected retrospectively. The clinical features and the results of mutational analysis are summarized in Table 4. Thirteen patients were responders who were shown partial or complete response. One patient (patient 11) showed the decrease of serum tumor marker (carcinoembryonic antigen) for 50% after gefitinib. This patient was considered as responder. Six patients were nonresponders who were stable disease or progressive disease. The mutations of the EGFR gene were detected in 11 of 14 (79%) responders, whereas none of six (0%) nonresponders had the mutation. The difference of the mutation rates between the responders and nonresponders was statistically significant (P = 0.0022). In patients 1 and 5, because tumors had the highly morphologic heterogeneity, we analyzed the EGFR mutation in two different portions in each sample. The identical mutations (delE746-A750 in patient 1 and L858R in patient 5) were found in each tumor. Totally, 11 mutations were found in 11 patients. Five were occurred around codons 746 to 750 in exon 19. Three were L858R in exon 21. These mutations were commonly found in our 120 fresh frozen tumor series and previous reports. We also found two missense mutations (N756D and G863D) and one silent mutation (L838L) in these patients.


View this table:
[in this window]
[in a new window]
  		Table 4. Characteristics of patients treated with gefitinib

 
Association EGFR mutations with KRAS mutations and RASSF1A hypermethylations in non–small cell lung cancers. Because EGFR plays an important role in Ras signal transduction, we analyzed the association EGFR mutation with KRAS mutation and hypermethylation of the RASSF1A gene, which has Ras association domain in 120 NSCLCs. Mutation of the KRAS gene was detected in 4 of 120 (3%) tumors, all of which had wild-type EGFR (Table 5). Methylation of the RASSF1A gene was detected in 35 (29%) tumors (26). EGFR mutation was detected in 13 of 35 (37%) tumors with RASSF1A methylations and detected in 16 of 85 (19%) tumors without RASSF1A methylations. The statistical significance of the correlation between EGFR mutation and RASSF1A methylation was borderline (P = 0.0587).


View this table:
[in this window]
[in a new window]
  		Table 5. Correlation of EGFR mutation with KRAS mutation and RASSF1A methylation in NSCLC

 

   Discussion
Top
 Abstract
 Materials and Methods
 Results
 Discussion
References
 
The frequency of EGFR mutations in NSCLCs were reported 26% to 40% in Asian patients and 2% to 12% in non-Asian patients, and almost all mutations were detected in adenocarcinoma (13–19). We first examined EGFR mutations in NSCLCs from 120 Japanese patients without gefitinib treatment. The frequency of the EGFR mutation was 24% and all mutations were found in adenocarcinoma. This result was almost concordant with previous studies. These results, including the present study, showed that EGFR mutation is actually high in Asian patients. Paez et al. (14) reported that EGFR mutations were more frequent in females than males. Pao et al. (15) reported that EGFR mutations were common in never smokers. It was also reported that EGFR mutations were more frequent in females and in never smokers (16, 18, 19). On the other hand, Huang et al. (17) reported that there was no significant association EGFR mutation with gender and smoking history. We found that mutations of the EGFR gene were predominantly detected in females and patients without smoking history. Previous three studies (14, 16, 19) and we analyzed the EGFR mutations in Japanese patients. On the other hand, Huang et al. (17) analyzed them in Taiwanese patients. These results suggested that although the frequency of the EGFR mutation is high in Asian patients, the features of the patients with the EGFR mutations are different in geographic distribution even in Asia.

We also found that EGFR mutation was more frequently detected in younger patients than older patients and in advanced-stage than in early-stage lung cancers. These results may suggest that EGFR mutation play a role in lung carcinogenesis in younger patients and in the progression of NSCLC. However, it was reported that there was no association EGFR mutation with age and stage of the disease (16). Because the result of the present study was discordant the previous study, further analyses will be needed to elucidate these issues.

We also showed here that EGFR mutations were correlated with the sensitivity to gefitinib. This result confirmed the previous studies (13–15, 17, 19). In the present study, we analyzed the two tumor samples obtained by the transbronchial lung biopsy. We could detect the EGFR mutation in one patient (patient 3), although we could not detect the mutation in another patient (patient 13) who was sensitive to gefitinib. One possibility is that there was actually no mutation of exons 18 to 21 of the EGFR gene in the tumor from the latter patient, although tumor was responded to gefitinib. The histology of the tumor of this patient was squamous cell carcinoma in which the EGFR mutation was very rare in both present and previous studies (13–17, 19). Another possibility is that the mutation could not be detected in this small sample obtained by transbronchial lung biopsy. One of the most characteristic features of lung cancers is the high degree of morphologic heterogeneity. We and others have reported that the morphologic changes of lung cancers are related to accumulation of genetic alterations (24, 27, 28). Therefore, the mutation could not be detected in this small sample by morphologic and genetic heterogeneity. However, we analyzed the EGFR mutation in two different portions in each of two samples (patients 1 and 5) that had the highly morphologic heterogeneity, and we found that the mutation was identical in the different morphologic portions. These results suggested that the examination of EGFR mutation could be available even in small samples obtained by biopsy such as transbronchial lung biopsy or percutaneous lung biopsy.

Minna et al. (29, 30) hypothesized that because RAS-mediated signal transduction lies downstream of EGFR, both of KRAS and EGFR genes are not required for lung cancer pathogenesis. Therefore, the mutations of these genes would be inversely correlated. It was reported that there was exclusionary relationship between KRAS mutation and EGFR mutation (16, 18). In the present study, although the difference between KRAS mutation and EGFR mutation was not statistically significant, there was no tumor that had mutations in both genes. This result could bear out their hypothesis as the previous study. The frequencies of KRAS mutation were 15% to 20% of all NSCLC and 20% to 30% of lung adenocarcinoma (31). In the present study, KRAS mutation was detected in 3% of NSCLC and 6% of adenocarcinoma. These frequencies were lower than that in the previous report. However, it has been reported that the frequencies of KRAS mutation were 4% to 10% of NSCLC and 8% to 13% of lung adenocarcinoma among the East Asian patients (16, 32–35). These results suggested that the high prevalence of EGFR mutation and the low prevalence of KRAS mutation are features in East Asian patients with lung adenocarcinoma. Furthermore, we analyzed the relationship between EGFR mutation and hypermethylation of the RASSF1A gene that has Ras association domain. We have reported that aberrant methylation of the RASSF1A gene predominantly occurs in adenocarcinoma (26). Recently, it was reported that RASSF1A knockout mice showed lung adenoma, breast adenocarcinoma, and lymphoma (36). In the present study, we found EGFR mutation tended to correlate with RASSF1A methylation. These results including the present results indicated that both EGFR mutation and RASSF1A methylation play a cooperative role in pathogenesis of lung adenocarcinoma.

In conclusion, EGFR mutation was frequently detected in Japanese patients with NSCLC, and all mutations were found in adenocarcinoma. EGFR mutation was predominantly detected in female, nonsmoker, younger patients, advanced-stage tumors, and well-differentiated tumors. Furthermore, EGFR mutation was significantly correlated with responsiveness to gefitinib. These results could assist to select patients who should be treated by gefitinib. However, because the present study was a retrospective analysis and because of the relatively small number of patients studied, prospective studies in a large number of patients will be required to be a diagnostic tool for selection of the treatment method based on molecular classification.


   Footnotes
 
Grant support: Ministry of Health, Labor, and Welfare; Ministry of Education, Culture, Sports, Science, and Technology, Japan grant-in-aid for scientific research grant 14570403-00, and Lung Cancer Specialized Programs of Research Excellence (USA) grants P50CA70907 and CA71618.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Note: Y. Tomizawa and H. Iijima contributed equally to the present study.

Received 2/28/05; revised 6/27/05; accepted 7/ 7/05.


   References
Top
 Abstract
 Materials and Methods
 Results
 Discussion
 References
 

  1. Rusch V, Baselga J, Cordon-Cardo C, et al. Differential expression of the epidermal growth factor receptor and its ligands in primary non-small cell lung cancers and adjacent benign lung. Cancer Res 1993;53:2379–85.[Abstract/Free Full Text]
  2. Fontanini G, De Laurentiis M, Vignati S, et al. Evaluation of epidermal growth factor-related growth factors and receptors and of neoangiogenesis in completely resected stage I-IIIA non-small-cell lung cancer: amphiregulin and microvessel count are independent prognostic indicators of survival. Clin Cancer Res 1998;4:241–9.[Abstract]
  3. Nicholson RI, Gee JM, Harper ME. EGFR and cancer prognosis. Eur J Cancer 2001;37:S9–15.
  4. Jorissen RN, Walker F, Pouliot N, Garrett TP, Ward CW, Burgess AW. Epidermal growth factor receptor: mechanisms of activation and signalling. Exp Cell Res 2003;284:31–53.[CrossRef][Medline]
  5. Olayioye MA, Neve RM, Lane HA, Hynes NE. The ErbB signaling network: receptor heterodimerization in development and cancer. EMBO J 2000;19:3159–67.[CrossRef][Medline]
  6. Pinkas-Kramarski R, Soussan L, Waterman H, et al. Diversification of Neu differentiation factor and epidermal growth factor signaling by combinatorial receptor interactions. EMBO J 1996;15:2452–67.[Medline]
  7. Druker BJ, Sawyers CL, Kantarjian H, et al. Activity of a specific inhibitor of the BCR-ABL tyrosine kinase in the blast crisis of chronic myeloid leukemia and acute lymphoblastic leukemia with the Philadelphia chromosome. N Engl J Med 2001;344:1038–42.[Abstract/Free Full Text]
  8. Demetri GD, von Mehren M, Blanke CD, et al. Efficacy and safety of imatinib mesylate in advanced gastrointestinal stromal tumors. N Engl J Med 2002;347:472–80.[Abstract/Free Full Text]
  9. Slamon DJ, Leyland-Jones B, Shak S, et al. Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med 2001;344:783–92.[Abstract/Free Full Text]
  10. Ciardiello F, Tortora G. A novel approach in the treatment of cancer: targeting the epidermal growth factor receptor. Clin Cancer Res 2001;7:2958–70.[Abstract/Free Full Text]
  11. Fukuoka M, Yano S, Giaccone G, et al. Multi-institutional randomized phase II trial of gefitinib for previously treated patients with advanced non-small-cell lung cancer (The IDEAL 1 Trial). J Clin Oncol 2003;21:2237–46.[Abstract/Free Full Text]
  12. Kris MG, Natale RB, Herbst RS, et al. Efficacy of gefitinib, an inhibitor of the epidermal growth factor receptor tyrosine kinase, in symptomatic patients with non-small cell lung cancer: a randomized trial. JAMA 2003;290:2149–58.[Abstract/Free Full Text]
  13. 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;350:2129–39.[Abstract/Free Full Text]
  14. Paez JG, Janne PA, Lee JC, et al. EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science 2004;304:1497–500.[Abstract/Free Full Text]
  15. Pao W, Miller V, Zakowski M, et al. EGF receptor gene mutations are common in lung cancers from "never smokers" and are associated with sensitivity of tumors to gefitinib and erlotinib. Proc Natl Acad Sci U S A 2004;101:13306–11.[Abstract/Free Full Text]
  16. Kosaka T, Yatabe Y, Endoh H, Kuwano H, Takahashi T, Mitsudomi T. Mutations of the epidermal growth factor receptor gene in lung cancer: biological and clinical implications. Cancer Res 2004;64:8919–23.[Abstract/Free Full Text]
  17. Huang SF, Liu HP, Li LH, et al. High frequency of epidermal growth factor receptor mutations with complex patterns in non-small cell lung cancers related to gefitinib responsiveness in Taiwan. Clin Cancer Res 2004;10:8195–203.[Abstract/Free Full Text]
  18. Marchetti A, Martella C, Felicioni L, et al. EGFR mutations in non-small-cell lung cancer: analysis of a large series of cases and development of a rapid and sensitive method for diagnostic screening with potential implications on pharmacologic treatment. J Clin Oncol 2005;23:857–65.[Abstract/Free Full Text]
  19. Tokumo M, Toyooka S, Kiura K, et al. The relationship between epidermal growth factor receptor mutations and clinicopathological features in non-small cell lung cancers. Clin Cancer Res 2005;11:1167–73.[Abstract/Free Full Text]
  20. Amann J, Kalyankrishna S, Massion PP, et al. Aberrant epidermal growth factor receptor signaling and enhanced sensitivity to EGFR inhibitors in lung cancer. Cancer Res 2005;65:226–35.[Abstract/Free Full Text]
  21. Kobayashi S, Boggon T, Dayaram T, et al. EGFR mutation and resistance of non-small cell lung cancer to gefitinib. N Engl J Med 2005;352:786–92.[Abstract/Free Full Text]
  22. Travis WD, Colby TV, Corrin B, Shimosato Y, Brambilla E. WHO International histological classification of tumors: histological typing of lung and pleural tumors. 3rd ed. Berlin: Springer; 1999.
  23. Yokota J, Wada M, Shimosato Y, Terada M, Sugimura T. Loss of heterozygosity on chromosomes 3, 13, and 17 in small-cell carcinoma and on chromosome 3 in adenocarcinoma of the lung. Proc Natl Acad Sci U S A 1987;84:9252–6.[Abstract/Free Full Text]
  24. Iijima H, Tomizawa Y, Dobashi K, Saito R, Nakajima T, Mori M. Allelic losses on chromosome 3p are accumulated in relation to morphological changes of lung adenocarcinoma. Br J Cancer 2004;91:1143–8.[Medline]
  25. Kohno T, Takakura S, Yamada T, Okamoto A, Tanaka T, Yokota J. Alterations of the PPP1R3 gene in human cancer. Cancer Res 1999;59:4170–4.[Abstract/Free Full Text]
  26. Tomizawa Y, Iijima H, Nomoto T, et al. Clinicopathological significance of aberrant methylation of RARß2 at 3p24, RASSF1A at 3p21.3, and FHIT at 3p14.2 in patients with non-small cell lung cancer. Lung Cancer 2004;46:305–12.[CrossRef][Medline]
  27. Yatabe Y, Konishi H, Mitsudomi T, Nakamura S, Takahashi T. Topographical distributions of allelic loss in individual non-small-cell lung cancers. Am J Pathol 2000;157:985–93.[Abstract/Free Full Text]
  28. Wistuba II, Behrens C, Virmani AK, et al. High resolution chromosome 3p allelotyping of human lung cancer and preneoplastic/preinvasive bronchial epithelium reveals multiple, discontinuous sites of 3p allele loss and three regions of frequent breakpoints. Cancer Res 2000;60:1949–60.[Abstract/Free Full Text]
  29. Minna JD, Gazdar AF, Sprang SR, Herz J. Cancer. A bull's eye for targeted lung cancer therapy. Science 2004;304:1458–61.[Abstract/Free Full Text]
  30. Gazdar AF, Shigematsu H, Herz J, Minna JD. Mutations and addiction to EGFR: the Achilles ‘heal’ of lung cancers? Trends Mol Med 2004;10:481–6.[CrossRef][Medline]
  31. Sekido Y, Fong KM, Minna JD. Progress in understanding the molecular pathogenesis of human lung cancer. Biochim Biophys Acta 1998;1378:F21–59.[Medline]
  32. Tomizawa Y, Kohno T, Kondo H, et al. Clinicopathological significance of epigenetic inactivation of RASSF1A at 3p21.3 in stage I lung adenocarcinoma. Clin Cancer Res 2002;8:2362–8.[Abstract/Free Full Text]
  33. Fukuyama Y, Mitsudomi T, Sugio K, Ishida T, Akazawa K, Sugimachi K. K-ras mutation and p53 mutations are an independent unfavourable prognostic indicator in patients with non-small-cell lung cancer. Br J Cancer 1997;75:1125–30.[Medline]
  34. Noda N, Matsuzoe D, Kohno T, Kawahara K, Yamashita Y, Shirakusa T. K-ras gene mutations in non-small cell lung cancer in Japanese. Oncol Rep 2001;8:889–92.[Medline]
  35. Soung YH, Lee JW, Kim SY, et al. Mutational analysis of EGFR gene and K-ras gene in lung adenocarcinoma. Virchows Arch 2005;446:483–8.[CrossRef][Medline]
  36. Tommasi S, Dammann R, Zhang Z, et al. Tumor susceptibility of Rassf1a knockout mice. Cancer Res 2005;65:92–8.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 J. Mol. Diagn.Home page
K. E. Finberg, L. V. Sequist, V. A. Joshi, A. Muzikansky, J. M. Miller, M. Han, J. Beheshti, L. R. Chirieac, E. J. Mark, and A. J. Iafrate
Mucinous Differentiation Correlates with Absence of EGFR Mutation and Presence of KRAS Mutation in Lung Adenocarcinomas with Bronchioloalveolar Features
J. Mol. Diagn., July 1, 2007; 9(3): 320 - 326.
[Abstract] [Full Text] [PDF]

Home page
 ChestHome page
D. S. Zander
Transbronchial Fine-Needle Aspiration, Epidermal Growth Factor Receptor Mutations, and the Scorpion's Touch
Chest, June 1, 2007; 131(6): 1619 - 1620.
[Full Text] [PDF]

Home page
 ChestHome page
A. Horiike, H. Kimura, K. Nishio, F. Ohyanagi, Y. Satoh, S. Okumura, Y. Ishikawa, K. Nakagawa, T. Horai, and M. Nishio
Detection of Epidermal Growth Factor Receptor Mutation in Transbronchial Needle Aspirates of Non-Small Cell Lung Cancer
Chest, June 1, 2007; 131(6): 1628 - 1634.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
J. Subramanian and R. Govindan
Lung Cancer in Never Smokers: A Review
J. Clin. Oncol., February 10, 2007; 25(5): 561 - 570.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
L. V. Sequist, D. W. Bell, T. J. Lynch, and D. A. Haber
Molecular Predictors of Response to Epidermal Growth Factor Receptor Antagonists in Non-Small-Cell Lung Cancer
J. Clin. Oncol., February 10, 2007; 25(5): 587 - 595.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
R. Rosell, M. Taron, N. Reguart, D. Isla, and T. Moran
Epidermal Growth Factor Receptor Activation: How Exon 19 and 21 Mutations Changed Our Understanding of the Pathway
Clin. Cancer Res., December 15, 2006; 12(24): 7222 - 7231.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
G. J. Riely, K. A. Politi, V. A. Miller, and W. Pao
Update on Epidermal Growth Factor Receptor Mutations in Non-Small Cell Lung Cancer
Clin. Cancer Res., December 15, 2006; 12(24): 7232 - 7241.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
A. Inoue, T. Suzuki, T. Fukuhara, M. Maemondo, Y. Kimura, N. Morikawa, H. Watanabe, Y. Saijo, and T. Nukiwa
Prospective Phase II Study of Gefitinib for Chemotherapy-Naive Patients With Advanced Non-Small-Cell Lung Cancer With Epidermal Growth Factor Receptor Gene Mutations
J. Clin. Oncol., July 20, 2006; 24(21): 3340 - 3346.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
E. L. Kwak, J. Jankowski, S. P. Thayer, G. Y. Lauwers, B. W. Brannigan, P. L. Harris, R. A. Okimoto, S. M. Haserlat, D. R. Driscoll, D. Ferry, et al.
Epidermal growth factor receptor kinase domain mutations in esophageal and pancreatic adenocarcinomas.
Clin. Cancer Res., July 15, 2006; 12(14): 4283 - 4287.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tomizawa, Y.
Right arrow Articles by Mori, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tomizawa, Y.
Right arrow Articles by Mori, M.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Cell Growth & Differentiation