Cancer Research CTRC-AACR San Antonio Breast Cancer Symposium AACR Conference on Molecular Diagnostics - 2008
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 Meeting Abstracts Online
[Cancer Research 59, 4798-4803, October 1, 1999]
© 1999 American Association for Cancer Research
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 Tseng, J. E.
Right arrow Articles by Mao, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tseng, J. E.
Right arrow Articles by Mao, L.
[Cancer Research 59, 4798-4803, October 1, 1999]
© 1999 American Association for Cancer Research

Advances in Brief

Loss of Fhit Is Frequent in Stage I Non-Small Cell Lung Cancer and in the Lungs of Chronic Smokers1

Jennifer E. Tseng, Bonnie L. Kemp, Fadlo R. Khuri, Jonathan M. Kurie, Jin Soo Lee, Xiao Zhou, Diane Liu, Waun Ki Hong and Li Mao2

Molecular Biology Laboratory at the Department of Thoracic/Head and Neck Medical Oncology [J. E. T., F. R. K., J. M. K., J. S. L., X. Z., D. L., W. K. H., L. M.] and the Department of Pathology [B. L. K.], the University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030


   ABSTRACT
Top
 ABSTRACT
Introduction
 Materials and Methods
 Results
 Discussion
 REFERENCES
 
Abnormalities of FHIT, a candidate tumor suppressor gene at 3p14.2, have been found frequently in multiple tumor types including non-small cell lung cancer (NSCLC). To investigate whether FHIT inactivation plays a role in early lung tumorigenesis, Fhit levels were determined by immunohistochemistry in tumors from 87 patients with stage I NSCLC and in 372 bronchial biopsy specimens from 86 chronic smokers without evidence of malignancy. We found that 49% of NSCLC specimens demonstrated significantly decreased staining or lack of staining for Fhit. However, Fhit expression status was not significantly associated with disease-free survival or overall survival. Analysis of a subset of 76 specimens on which microsatellite analysis at the FHIT locus was performed did not show a strong association between loss of heterozygosity at FHIT and Fhit expression, suggesting the presence of complex mechanisms of Fhit inactivation. Of 372 bronchial biopsies from chronic smokers, 86 biopsies (23%) exhibited decreased Fhit expression or lack of Fhit expression. In 37 of 86 (43%) subjects, decreased Fhit expression or lack of expression was observed in at least one biopsy site. Loss of Fhit expression was significantly higher in bronchial metaplastic lesions (23 of 49 lesions, 47%) than in histologically normal bronchial epithelium (63 of 323 specimens, 20%; P < 0.001). Smokers with a metaplasia index of >15% had a higher frequency of loss of Fhit expression than those with a metaplasia index of <=15% (P = 0.015). Interestingly, current smokers had a higher rate of loss of Fhit expression than former smokers (P = 0.02). Our data indicate that Fhit expression is significantly reduced in a substantial number of early-stage NSCLC and preneoplastic lesions in chronic smokers. The association between cigarette smoking and Fhit expression suggests a role for FHIT in the initiation of smoking-related lung tumorigenesis.


   Introduction
Top
 ABSTRACT
 Introduction
Materials and Methods
 Results
 Discussion
 REFERENCES
 
Lung cancer remains the leading cause of cancer-related mortality in the United States (1) . It was estimated that 171,600 new cases of lung cancer would be diagnosed in 1998, with an estimated 158,900 deaths (1) . The 5-year-survival rate of 14% for patients with lung cancer has improved little over the past decade (1) . Even patients with pathological stage I NSCLC3 have only a 60% survival rate at 5 years. Two concepts, field cancerization and multistep tumorigenesis, are essential in understanding the underlying mechanisms of carcinogenesis of the aerodigestive tract. Field cancerization was first proposed by Slaughter and Skejkal in 1953 (2) , who suggested that a whole tissue field exposed to common carcinogens, such as cigarette smoke, is at risk for the development of malignancy due to diffuse injury over time. Multistep tumorigenesis refers to the multistep process in which genetic events accumulate and result in malignant transformation, as well illustrated in the colorectal cancer model (3) .

It is estimated that between 10 and 20 genetic events are required for lung tumorigenesis (4) . Numerous genetic alterations have been identified that occur at high frequencies in NSCLC. These include p53 gene mutations (5 , 6) , K-ras proto-oncogene mutations (7) , inactivation of the Rb gene (8) , and alterations in p16, CDC25, cyclin D1, and FHIT (9, 10, 11, 12, 13) . The FHIT gene is located at 3p14.2 (14) , a region frequently lost in multiple tumor types. Loss of Fhit expression has been found to occur frequently in multiple tumor types including NSCLC (10 , 15) . To determine whether loss of Fhit occurs early and plays an important role in lung tumorigenesis, we analyzed Fhit expression in primary tumors from 87 patients with stage I NSCLC and in 372 bronchial biopsy specimens from 86 chronic smokers without evidence of malignancy. We found that Fhit expression is absent or markedly reduced in 49% of 87 primary tumors in patients with stage I NSCLC and in 23% of 372 bronchial biopsy specimens from chronic smokers. We also found that loss of Fhit expression was associated with bronchial metaplastic changes, patients’ MI, and current smoking status.


   Materials and Methods
Top
 ABSTRACT
 Introduction
 Materials and Methods
Results
 Discussion
 REFERENCES
 
Study Population.
Primary NSCLC specimens were obtained from archived tissue specimens of surgically resected pathological stage I tumors from 87 patients treated at the M. D. Anderson Cancer Center (Houston, TX). Clinical and pathological features of the study population are listed in Table 1Citation . Surgical specimens were collected between 1975 and 1990. Survival data were available for all patients for at least 5 years. The study population consisted of 62 males and 25 females. The mean age of patients was 64.0 years. Histological subtypes included 37 adenocarcinomas, 36 squamous cell carcinomas, 8 bronchoalveolar carcinomas, 3 large cell carcinomas, 1 adenosquamous carcinoma, and 2 unclassified tumors.


View this table:
[in this window]
[in a new window]
  		Table 1 Fhit reactivity in stage I NSCLC tumors according to clinicopathological features of patients

 
A total of 372 bronchial biopsies were obtained from 86 chronic smokers (72 current and 14 former smokers) who were enrolled in a chemoprevention protocol at the M. D. Anderson Cancer Center between 1993 and 1998, before any intervention. Chronic smokers were defined as subjects who had smoked >=1 pack/day for >=20 years and who were currently smoking at the time of enrollment in the study or may have quit at any time before enrollment. Current smokers were defined as subjects who were smoking at the time of enrollment or who had stopped smoking within a year before enrollment. Former smokers were defined as subjects who had stopped smoking >12 months before enrollment. Thirty-eight percent (33 of 86) were female, and 62% (53 of 86) were male. The median number of pack-years for all subjects was 30.5; the median number of packs/day was 1.2. The median age of subjects was 54 years. All patients underwent fiber-optic bronchoscopy. Random endobronchial biopsies were obtained from six predetermined sites, including the right upper lobe bronchus, the right middle lobe bronchus, the right lower lobe bronchus, the left upper lobe bronchus, the left lower lobe bronchus, and the carina. Forty-nine of 372 specimens contained bronchial metaplasia; the remainder did not contain any histological abnormalities. The MI was calculated for each subject; the MI was defined as the number of biopsies exhibiting metaplasia divided by the total number of biopsies, with the quotient multiplied by 100%.

Immunohistochemical Staining for Fhit Protein.
Paraffin-embedded, 4-µm-thick tissue sections from all 87 primary tumors and 372 bronchial epithelial biopsies from 86 chronic smokers were stained for the Fhit protein using a primary rabbit polyclonal anti-glutathione S-transferase-Fhit antibody (kindly provided by Drs. Carlo Croce and Kay Huebner, Kimmel Cancer Center, Philadelphia, PA). Deparaffinization of all sections was performed through a series of xylene baths, and rehydration was performed through graded alcohols. Slides were incubated for 15 min in a 0.3% hydrogen peroxide-methanol solution to reduce endogenous peroxidase activity, followed by incubation in 2.5% normal goat serum for 30 min to reduce nonspecific binding. Slides were incubated overnight at room temperature with primary anti-glutathione S-transferase-Fhit antiserum (1:5000), followed by incubation for 30 min with biotinylated antirabbit IgG (Vector Laboratories, Burlingame, CA). Antibody localization was performed using the manufacturer’s protocol (Vector Laboratories). Normal bronchial tissue sections from a nonsmoker were used as positive controls for each batch of staining for both the NSCLC specimens and the bronchial biopsies.

A four-tiered scoring system was used that incorporates both intensity of staining (absent, 0; low level, 1; moderate, 2; and strong, 3) and the percentage of cells stained (<5%, 1; 5–50%, 2; and >50%, 3). A composite score of 0–9 for the Fhit protein level was obtained by multiplying the intensity and extent scores for each tissue section. Composite scores <= 3 were defined as a marked reduction or absence of Fhit protein expression, whereas composite scores of >3 were considered positive for Fhit protein expression. All slides were scored concomitantly by one pathologist (B. L. K.) and another investigator (J. E. T.), who reached agreement regarding both the intensity and percentage of staining for each tissue section.

DNA Extraction and Microsatellite Analysis.
Microsatellite analysis was performed on a subset of 76 cases of NSCLC for which immunohistochemical analysis had been performed. Sections adjacent to those used for immunohistochemistry were deparaffinized and stained with H&E. Tumor cells and adjacent bronchial alveolar normal cells were microdissected from each section using a 25.5-gauge steel needle under a stereomicroscope. Approximately 1000–2000 cells were dissected from each section. After microdissection, DNA was extracted as described previously (16) . The markers used for microsatellite analysis were D3S1234 and D3S1481 on 3p14 within the FHIT gene (Research Genetics, Huntsville, AL). Microsatellite analysis was performed as described previously (16) . LOH was defined as a >50% decrease in signal intensity by visual inspection in one of the two alleles compared to that in the corresponding normal control. The results were interpreted by two independent observers (J. E. T. and L. M.), and agreement was reached after discussion for any with discrepancies. If either of the two markers demonstrated LOH, then the patient was considered to exhibit LOH at the FHIT locus.

Statistical Analysis.
Survival curves were estimated by the Kaplan-Meier method, and the resulting curves were compared using the log-rank test. Fisher’s exact test and {chi}2 test were used to analyze the association between two categorical variables. P < 0.05 was considered to be statistically significant. Immunohistochemical analysis was performed in a blinded manner with respect to the clinical information for the subjects, both for the stage I NSCLC patients and for the chronic smokers.


   Results
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results
Discussion
 REFERENCES
 
Loss of Fhit Expression in Stage I NSCLC.
Immunohistochemical staining was performed on 87 tumor sections from 87 stage I NSCLC cases. In normal bronchial tissues, cytoplasmic granular staining of Fhit was observed in bronchial epithelial cells (Fig. 1, A and B)Citation . We found that 49% (43 of 87) of pathological stage I NSCLC tissue sections showed decreased or negative staining for Fhit (score, <=3), whereas 51% of NSCLC tissue sections showed at least moderate levels of staining (score, >3). The scoring system we applied is similar to that used by Greenspan et al. (17) and Hadaczek et al. (18) using the same antibody. Other studies have used a scoring system that classified tumors as positive, mixed, or negative for Fhit immunoreactivity (19 , 20) . We did observe a mixed pattern in many tumors and bronchial epithelial specimens with different staining intensities within the same section; thus, we used an immunoreactivity score that takes into account both the intensity of cells stained and the percentage of cells stained. In tumors and epithelial specimens displaying a mixed pattern of staining, the highest intensity of staining within the tumor was used as the intensity score. Examples of Fhit expression patterns in stage I NSCLC primary tumors are shown in Fig. 1Citation . It should be noted that the Fhit positive category represents a range of intensities and extents of Fhit expression, many of which may be consistent with LOH.



View larger version (177K):
[in this window]
[in a new window]
  		Fig. 1. Fhit immunohistochemical staining in normal bronchial tissue and NSCLC primary tumors. a and b, normal bronchial epithelium (arrows) with strongly positive immunostaining. c, Fhit-positive immunostaining in a bronchoalveolar carcinoma (arrow) demonstrating strong staining intensity in >50% of tumor cells. d, weak immunostaining for Fhit in >50% tumor cells in an adenocarcinoma (arrow). e, squamous cell carcinoma (arrow) demonstrating moderate immunostaining for Fhit in >50% tumor cells. f, squamous cell carcinoma (arrow) demonstrating negative immunostaining for Fhit in >50% tumor cells.

 
The frequency of Fhit negativity in patients with stage I NSCLC did not differ significantly according to gender, age, histological subtype, or smoking status (Table 1)Citation . However, it should be noted that the comparison between smokers and nonsmokers is limited by the small sample size of the nonsmoker group (Table 1)Citation . OS and DFS did not differ significantly between patients with Fhit-negative and Fhit-positive tumors (Table 1)Citation . In the squamous cell carcinoma and adenocarcinoma subsets, OS and DFS did not differ significantly between patients with Fhit-negative and Fhit-positive tumors.

Loss of Fhit Expression in Stage I NSCLC and LOH at the FHIT Gene.
Microsatellite analysis was performed on 76 of 87 tissue sections on which immunohistochemical protein analysis was also performed. We were unable to perform microsatellite analysis on 11 of 87 tissue sections due to lack of tissue samples, lack of reliable normal tissues as controls, or the poor quality of DNA. The two highly polymorphic markers, D3S1234 and D3S1481, are located within the FHIT gene at the 3p14 locus; D3S1234 is located in intron 5 (21) , and D3S1481 is located in intron 4. Fifty-nine of 76 (78%) and 44 of 76 (58%) cases were informative at the D3S1234 and D3S1481 markers, respectively; 67 of 76 (88%) cases were informative at one or both markers. LOH at markers D3S1234 and D3S1481 were observed in 34 of 59 (58%) and 15 of 44 (34%) informative cases, respectively. Thirty-nine of 67 (58%) informative cases demonstrated LOH in at least one marker. Twenty-three of 36 (64%) tumors with negative Fhit expression demonstrated LOH at the FHIT locus, whereas 16 of 31 (52%) tumors with positive Fhit expression demonstrated LOH at the FHIT locus. Thus, a slightly higher proportion of tumors with loss of Fhit expression demonstrated LOH at FHIT compared with tumors with normal Fhit expression, although this difference was not statistically significant (P = 0.31, {chi}2 test).

Loss of Fhit Expression in Bronchial Epithelial Specimens from Chronic Smokers.
Loss of Fhit expression was observed in 86 of 372 (23%) bronchial epithelial biopsy specimens, including 63 of 323 (20%) bronchial epithelial biopsy specimens without metaplasia, and in 23 of 49 (47%) biopsy specimens containing bronchial metaplasia (P < 0.001; Table 2Citation ; Fig. 2Citation ). Loss of Fhit expression in the bronchial epithelium in at least one biopsy site was demonstrated in 37 of 86 cases (43%), including 8 of 31 (26%) of the cases with a metaplasia index of <=15% and in 29 of 55 (53%) of the cases with a metaplasia index of >15% (P = 0.015).


View this table:
[in this window]
[in a new window]
  		Table 2 Fhit reactivity in bronchial epithelial specimens in chronic smokers

 


View larger version (107K):
[in this window]
[in a new window]
  		Fig. 2. Fhit immunohistochemical staining in histologically normal bronchial epithelium and bronchial metaplasia from chronic smokers. a, strong Fhit immunostaining in histologically normal bronchial epithelium (arrow). b, weak Fhit immunostaining in histologically normal bronchial epithelium (arrow). c, strong Fhit immunostaining in a bronchial metaplastic lesion (arrow). d, weak Fhit immunostaining in a bronchial metaplastic lesion (arrow).

 
Interestingly, all biopsy specimens containing metaplasia were from current smokers. In bronchial biopsies without metaplasia, loss of Fhit expression was observed in only 4 of 59 (7%) biopsy sites from former smokers and 59 of 264 (22%) biopsy sites from current smokers (P = 0.006, Fisher’s exact test), compared to 23 of 49 (47%) biopsy sites with metaplasia from current smokers. When analyzed by case, loss of Fhit expression was observed in at least one biopsy specimen in 35 of 72 (49%) current smokers compared to 2 of 14 (14%) biopsy specimens from former smokers (P = 0.02, Fisher’s exact test).


   Discussion
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results
 Discussion
REFERENCES
 
The role of the FHIT gene as a tumor suppressor gene has been controversial. Tumor suppressor genes may be inactivated by several mechanism, including intragenic deletions, point mutations, loss of a whole chromosome, or genetic recombination (22) . One feature of FHIT is the virtual absence of point mutations (23 , 24) . Furthermore, a number of investigators have reported the coexistence of wild-type transcripts along with aberrant FHIT transcripts in tumors (10 , 14 , 23) . Additionally, aberrant FHIT transcripts have been found in nonmalignant tissues (25, 26, 27) . However, multiple lines of evidence suggest that FHIT is a tumor suppressor gene. First, FHIT is located in a region of the genome that is known to be rearranged and/or deleted in multiple tumor types (10 , 14 , 28, 29, 30) . Homozygous deletions of the genetic region have been identified in multiple types of tumors; in some cases, the deletions included FHIT exons (14 , 31) . Furthermore, aberrant FHIT transcripts and/or decreased protein expression have been identified in multiple tumor types, including renal cell, cervical, and lung cancers (15 , 17, 18, 19, 20) . It has been demonstrated that transfection of wild-type FHIT genes into cancer cell lines lacking FHIT suppressed tumorigenicity in nude mice (32) . More recently, Ji et al. (33) found that overexpression of the FHIT gene in lung and head and neck cancer cell lines induced apoptosis, caused cell cycle arrest in S phase, and suppressed tumorigenicity.

We observed frequent loss of Fhit protein expression in early-stage NSCLC, with 49% of specimens demonstrating a decrease or lack of Fhit protein staining. This frequency of decreased or absent protein expression is similar to that observed in NSCLC by other investigators (15 , 19 , 20) . However, we did not find a significant association between the lack of Fhit expression and survival in stage I NSCLC. This is consistent with a recent report by Sozzi et al. (19) . However, Tomizawa et al. (20) found that lack of Fhit expression in Japanese patients with NSCLC predicted a poorer outcome. The lack of correlation between Fhit protein inactivation and survival in our study and in that of Sozzi et al. (19) may be due to a role of FHIT at an early stage in lung carcinogenesis, whereas different scoring methodologies and patient populations may account in part for the discordance between our results and those of Tomizawa et al. (20) . Additionally, the latter study included a larger proportion of adenocarcinomas, a factor that may have contributed to the different outcome of a subset of patients in this study (20) .

A number of studies have found that alterations in the 3p locus occur frequently in bronchial preneoplastic lesions (34 , 35) from chronic smokers. Sozzi et al. (19) observed loss of Fhit expression in 17 of 20 (85%) dysplastic lesions and in 25 of 25 (100%) carcinoma in situ lesions at the margins of tumor specimens from patients with lung cancer, further suggesting that inactivation of the FHIT gene may occur at an early stage in lung carcinogenesis. In this study, we have analyzed Fhit expression status in a large number of bronchial biopsies from chronic smokers. We have found that epithelium in 23% of the biopsies from 43% of smokers showed a lack of Fhit expression or a significantly reduced Fhit expression. Loss of Fhit expression was significantly more frequent in metaplastic lesions than in histologically normal epithelium (47% versus 20%, respectively) in chronic smokers. To our knowledge, our study is the first to demonstrate loss of Fhit expression in histologically normal bronchial epithelium and frequent loss of Fhit expression in bronchial metaplastic lesions from chronic smokers. Furthermore, loss of Fhit expression was more frequent in smokers with a high MI (>15%) than in smokers with a MI <= 15% (53% versus 26%). We have previously demonstrated that LOH at 3p14 occurs frequently in the histologically normal bronchial epithelium of chronic smokers (35) . Together, these observations strongly support the notion that loss of Fhit expression occurs early in the progression from normal to premalignant lesions in the lungs of smokers. Furthermore, loss of Fhit expression appears to be strongly associated with current smoking status, suggesting that the FHIT gene may be the target of carcinogens in cigarette smoke. These data further support a potential role of the FHIT gene in smoking-related lung carcinogenesis. Recent epidemiological studies indicate that former smokers account for over 50% of lung cancer cases; former smokers have over twice the lung cancer risk of lifetime nonsmokers (36, 37, 38) . These data underscore the importance of investigating the genetic abnormalities associated with smoking.

In this study, we did not find a concordance between LOH at the FHIT gene and loss of Fhit expression in NSCLC. Whereas LOH is generally associated with decreased protein expression, protein inactivation may be due to complex mechanisms other than deletion within an allele. Tanaka et al. (39) investigated such mechanisms of FHIT inactivation in esophageal cell lines and tumors and found hypermethylation of the 5' CpG island to be an important mechanism for FHIT inactivation in some cell lines. Alternatively, abnormal protein expression may occur in the absence of genetic alterations through altered splicing fidelity (27) . Additionally, posttranslational events such as ubiquitin-proteosome-dependent degradation (40) have been found to be important mechanisms of protein inactivation for other genes; however, such mechanisms have not previously been investigated for Fhit. It is also possible that microsatellite analysis may not be a sensitive method to detect homozygous deletions within the FHIT gene that have been reported in NSCLC cell lines as a mechanism of FHIT inactivation (41 , 42) . In fact, homozygous deletions in the FRA3B region located within the FHIT gene have been reported in certain NSCLC cell lines (41 , 42) .

Additionally, the scoring system that we and others have used for protein expression incorporates both the intensity of staining and the percentage of cells stained and thus includes heterogenous patterns of immunoreactivity within subsets of tumors that are classified as positive and those that are classified as negative. A tumor with an immunoreactivity score of 6 might have strong staining in a minority of cells (and perhaps loss of both alleles in most cells) or moderate expression in a majority of cells (and perhaps loss of one allele in most cells). Thus, LOH data do not provide information about mechanisms of protein inactivation that do not involve deletion.

In summary, we have demonstrated that Fhit protein inactivation is a frequent event in bronchial premalignant lesions and in stage I NSCLC, suggesting that loss of Fhit expression occurs early in the progression from normal to malignant lesions. Importantly, we found strong associations between loss of Fhit expression and cigarette smoking, metaplastic changes, and high MI, suggesting a potential role for FHIT in smoking-related lung tumorigenesis.


   ACKNOWLEDGMENTS
 
We thank Susan Cweren, Lakshmi Kakarala, and Shyla Kalapurakai for technical assistance with immunohistochemistry and assistance with the preparation of pathology slides.


   FOOTNOTES
 
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 with 18 U.S.C. Section 1734 solely to indicate this fact.

1 Supported in part by American Cancer Society Grant RPG-98-054 (to L. M.), American Cancer Society-Clinical Oncology Career Development Award 96-41 (to F. R. K.), M. D. Anderson Trainee and Student Resources Research Grant (to J. E. T.), National Cancer Institute No1 Grant (to W. K. H.), U19 CA 68437 Grant (to W. K. H.), and National Cancer Institute Cancer Center Grant P30 CA16620 (to the M. D. Anderson Cancer Center). W. K. H. is an American Cancer Society Clinical Research Professor. Back

2 To whom requests for reprints should be addressed, at Molecular Biology Laboratory at the Department of Thoracic/Head and Neck Medical Oncology, The University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030. Phone: (713) 792-6363; Fax: (713) 796-8865; Back

3 The abbreviations used are: NSCLC, non-small cell lung cancer; LOH, loss of heterozygosity; DFS, disease-free survival; OS, overall survival; MI, metaplasia index. Back

Received 6/29/99. Accepted 8/18/99.


   REFERENCES
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results
 Discussion
 REFERENCES
 

  1. Landis S. H., Murray T., Bolden S., Wingo P. A. Cancer statistics, 1999. CA Cancer J. Clin., 49: 8-31, 1999.[Abstract/Free Full Text]
  2. Slaughter M., Skejkal W. A. Field cancerization in oral stratified epithelium: clinical implications of multicentric origin. Cancer (Phila.), 6: 963-968, 1953.[Medline]
  3. Kinzler K. W., Vogelstein B. Lessons from hereditary colorectal cancer. Cell, 87: 159-171, 1996.[Medline]
  4. Minna J. D. Genetic events in the pathogenesis of lung cancer. Chest, 96: 17S-23S, 1989.[Free Full Text]
  5. Mitsudomi T., Steinberg S. M., Nau M. M., Carbone D., D’Amico D., Bodner S., Oie H. K., Linnoila R. I., Mulshine J. L., Minna J. D., Gazdar A. F. p53 gene mutations in non-small-cell lung cancer cell lines and their correlation with the presence of ras mutations and clinical features. Oncogene, 7: 171-180, 1992.[Medline]
  6. Hollstein M., Sidransky D., Vogelstein B., Harris C. C. p53 mutations in human cancers. Science (Washington DC), 253: 49-53, 1991.[Abstract/Free Full Text]
  7. Rosell R., Monzo M., Molina F., Martinez E., Pifarre A., Moreno I., Mate J. L., de Anta J. M., Sanchez M., Font A. K-ras genotypes and prognosis in non-small-cell lung cancer. Ann. Oncol., 6: S15-S20, 1995.
  8. Reissmann P. T., Koga H., Takahashi R., Figlin R. A., Holmes E. C., Piantadosi S., Cordon-Cardo C., Slamon D. J. Inactivation of the retinoblastoma susceptibility gene in non-small-cell lung cancer. The Lung Cancer Study Group. Oncogene, 8: 1913-1919, 1993.[Medline]
  9. Wu W., Fan Y. H., Kemp B. L., Walsh G., Mao L. Overexpression of cdc25A and cdc25B is frequent in primary non-small cell lung cancer but is not associated with overexpression of c-myc. Cancer Res., 58: 4082-4085, 1998.[Abstract/Free Full Text]
  10. Sozzi G., Veronese M. L., Negrini M., Baffa R., Cotticelli M. G., Inoue H., Tornielli S., Pilotti S., De Gregorio L., Pastorino U., Pierotti M. A., Ohta M., Huebner K., Croce C. M. The FHIT gene 3p14.2 is abnormal in lung cancer. Cell, 85: 17-26, 1996.[Medline]
  11. Merlo A., Herman J. G., Mao L., Lee D. J., Gabrielson E., Burger P. C., Baylin S. B., Sidransky D. 5' CpG island methylation is associated with transcriptional silencing of the tumour suppressor p16/CDKN2/MTS1 in human cancers. Nat. Med., 1: 686-692, 1995.[Medline]
  12. Marchetti A., Doglioni C., Barbareschi M., Buttitta F., Pellegrini S., Gaeta P., La Rocca R., Merlo G., Chella A., Angeletti C. A., Dalla Palma P., Bevilacqua G. Cyclin D1 and retinoblastoma susceptibility gene alterations in non-small cell lung cancer. Int. J. Cancer, 75: 187-192, 1998.[Medline]
  13. Cairns P., Polascik T. J., Eby Y., Tokino K., Califano J., Merlo A., Mao L., Herath J., Jenkins R., Westra W., et al Frequency of homozygous deletion at p16/CDKN2 in primary human tumours. Nat. Genet., 11: 210-212, 1995.[Medline]
  14. Ohta M., Inoue H., Cotticelli M. G., Kastury K., Baffa R., Palazzo J., Siprashvili Z., Mori M., McCue P., Druck T., et al The FHIT gene, spanning the chromosome 3p14.2 fragile site and renal carcinoma-associated t(3:8) breakpoint, is abnormal in digestive tract cancers. Cell, 84: 587-597, 1996.[Medline]
  15. Sozzi G., Tornielli S., Tagliabue E., Sard L., Pezzella F., Pastorino U., Minoletti F., Pilotti S., Ratcliffe C., Veronese M. L., Goldstraw P., Huebner K., Croce C. M., Pierotti M. A. Absence of Fhit protein in primary lung tumors and cell lines with FHIT gene abnormalities. Cancer Res., 57: 5207-5212, 1997.[Abstract/Free Full Text]
  16. Mao L., Lee J. S., Fan Y. H., Ro J. Y., Batsakis J. G., Lippman S., Hittelman W., Hong W. K. Frequent microsatellite alterations at chromosomes 9p21 and 3p14 in oral premalignant lesions and their value in cancer risk assessment. Nat. Med., 2: 682-685, 1996.[Medline]
  17. Greenspan D. L., Connolly D. C., Wu R., Lei R. Y., Vogelstein J. T., Kim Y. T., Mok J. E., Munoz N., Bosch F. X., Shah K., Cho K. R Loss of FHIT expression in cervical carcinoma cell lines and primary tumors. Cancer Res., 57: 4692-4698, 1997.[Abstract/Free Full Text]
  18. Hadaczek P., Siprashvili Z., Markiewski M., Domagala W., Druck T., McCue P. A., Pekarsky Y., Ohta M., Huebner K., Lubinski J. Absence or reduction of Fhit expression in most clear cell renal carcinomas. Cancer Res., 58: 2946-2951, 1998.[Abstract/Free Full Text]
  19. Sozzi G., Pastorino U., Moiraghi L., Tagliabue E., Pezzella F., Ghirelli C., Tornielli S., Sard L., Huebner K., Pierotti M. A., Croce C. M., Pilotti S. Loss of FHIT function in lung cancer and preinvasive bronchial lesions. Cancer Res., 58: 5032-5037, 1998.[Abstract/Free Full Text]
  20. Tomizawa Y., Nakajima T., Kohno T., Saito R., Yamaguchi N., Yokota J. Clinicopathological significance of Fhit protein expression in stage I non-small cell lung carcinoma. Cancer Res., 58: 5478-5483, 1998.[Abstract/Free Full Text]
  21. Mimori K., Druck T., Inoue H., Alder H., Berk L., Mori M., Huebner K., Croce C. M. Cancer-specific chromosome alterations in the constitutive fragile region FRA3B. Proc. Natl. Acad. Sci. USA, 96: 7456-7461, 1999.[Abstract/Free Full Text]
  22. Knudson A. G., Jr. Retinoblastoma: a prototypic hereditary neoplasm. Semin. Oncol., 5: 57-60, 1978.[Medline]
  23. Fong K. M., Biesterveld E. J., Virmani A., Wistuba I., Sekido Y., Bader S. A., Ahmadian M., Ong S. T., Rassool F. V., Zimmerman P. V., Giaccone G., Gazdar A. F., Minna J. D. FHIT and FRA3B 3p14.2 allele loss are common in lung cancer and preneoplastic bronchial lesions and are associated with cancer-related FHIT cDNA splicing aberrations. Cancer Res., 57: 2256-2267, 1997.[Abstract/Free Full Text]
  24. Nelson H. H., Wiencke J. K., Gunn L., Wain J. C., Christiani D. C., Kelsey K. T. Chromosome 3p14 alterations in lung cancer: evidence that FHIT exon deletion is a target of tobacco carcinogens and asbestos. Cancer Res., 58: 1804-1807, 1998.[Abstract/Free Full Text]
  25. Panagopoulos I., Thelin S., Mertens F., Mitelman F., Aman P. Variable FHIT transcripts in non-neoplastic tissues. Genes Chromosomes Cancer, 19: 215-219, 1997.[Medline]
  26. Chen Y. J., Chen P. H., Lee M. D., Chang J. G. Aberrant FHIT transcripts in cancerous and corresponding non-cancerous lesions of the digestive tract. Int. J. Cancer, 72: 955-958, 1997.[Medline]
  27. Gayther S. A., Barski P., Batley S. J., Li L., de Foy K. A., Cohen S. N., Ponder B. A., Caldas C. Aberrant splicing of the TSG101 and FHIT genes occurs frequently in multiple malignancies and in normal tissues and mimics alterations previously described in tumours. Oncogene, 15: 2119-2126, 1997.[Medline]
  28. Mao L., Fan Y. H., Lotan R., Hong W. K. Frequent abnormalities of FHIT, a candidate tumor suppressor gene, in head and neck cancer cell lines. Cancer Res., 56: 5128-5131, 1996.[Abstract/Free Full Text]
  29. Druck T., Kastury K., Hadaczek P., Podolski J., Toloczko A., Sikorski A., Ohta M., LaForgia S., Lasota J., McCue P., Lubinski J., Huebner K. Loss of heterozygosity at the familial RCC t(3:8) locus in most clear cell renal carcinomas. Cancer Res., 55: 5348-5353, 1995.[Abstract/Free Full Text]
  30. Virgilio L., Shuster M., Gollin S. M., Veronese M. L., Ohta M., Huebner K., Croce C. M. FHIT gene alterations in head and neck squamous cell carcinomas. Proc. Natl. Acad. Sci. USA, 93: 9770-9775, 1996.[Abstract/Free Full Text]
  31. Druck T., Hadaczek P., Fu T. B., Ohta M., Siprashvili Z., Baffa R., Negrini M., Kastury K., Veronese M. L., Rosen D., Rothstein J., McCue P., Cotticelli M. G., Inoue H., Croce C. M., Huebner K. Structure and expression of the human FHIT gene in normal and tumor cells. Cancer Res., 57: 504-512, 1997.[Abstract/Free Full Text]
  32. Siprashvili Z., Sozzi G., Barnes L. D., McCue P., Robinson A. K., Eryomin V., Sard L., Tagliabue E., Greco A., Fusetti L., Schwartz G., Pierotti M. A., Croce C. M., Huebner K. Replacement of Fhit in cancer cells suppresses tumorigenicity. Proc. Natl. Acad. Sci. USA, 94: 13771-13776, 1997.[Abstract/Free Full Text]
  33. Ji L., Fang B., Yen N., Fong K., Minna J. D., Roth J. A. Induction of apoptosis, inhibition of tumorigenicity, and suppression of tumor growth by adenovirus vector-mediated FHIT gene overexpression in vitro and in vivo. Cancer Res., 59: 3333-3339, 1999.[Abstract/Free Full Text]
  34. Wistuba I. I., Lam S., Behrens C., Virmani A. K., Fong K. M., LeRiche J., Samet J. M., Srivastava S., Minna J. D., Gazdar A. F. Molecular damage in the bronchial epithelium of current and former smokers. J. Natl. Cancer Inst., 89: 1366-1373, 1997.[Abstract/Free Full Text]
  35. Mao L., Lee J. S., Kurie J. M., Fan Y. H., Lippman S. M., Lee J. J., Ro J. Y., Broxson A., Yu R., Morice R. C., Kemp B. L., Khuri F. R., Walsh G. L., Hittelman W. N., Hong W. K. Clonal genetic alterations in the lungs of current and former smokers. J. Natl. Cancer Inst., 89: 857-862, 1997.[Abstract/Free Full Text]
  36. Huebner K., Garrison P. N., Barnes L. D., Croce C. M. The role of the FHIT/FRA3B locus in cancer. Annu. Rev. Genet., 32: 7-31, 1998.[Medline]
  37. Alavanja M. C., Brownson R. C., Benichou J., Swanson C., Boice J. D., Jr. Attributable risk of lung cancer in lifetime nonsmokers and long-term ex-smokers (Missouri, United States). Cancer Causes Control, 6: 209-216, 1995.[Medline]
  38. Tong L., Spitz M. R., Fueger J. J., Amos C. A. Lung carcinoma in former smokers. Cancer (Phila.), 78: 1004-1010, 1996.[Medline]
  39. Tanaka H., Shimada Y., Harada H., Shinoda M., Hatooka S., Imamura M., Ishizaki K. Methylation of the 5' CpG island of the FHIT gene is closely associated with transcriptional inactivation in esophageal squamous cell carcinomas. Cancer Res., 58: 3429-3434, 1998.[Abstract/Free Full Text]
  40. Pagano M., Tam S. W., Theodoras A. M., Beer-Romero P., Del Sal G., Chau V., Yew P. R., Draetta G. F., Rolfe M. Role of the ubiquitin-proteasome pathway in regulating abundance of the cyclin-dependent kinase inhibitor p27. Science (Washington DC), 269: 682-685, 1995.[Abstract/Free Full Text]
  41. Ong S. T., Fong K. M., Bader S. A., Minna J. D., Le Beau M. M., McKeithan T. W., Rassool F. V. Precise localization of the FHIT gene to the common fragile site at 3p14.2 (FRA3B) and characterization of homozygous deletions within FRA3B that affect FHIT transcription in tumor cell lines. Genes Chromosomes Cancer, 20: 16-23, 1997.[Medline]
  42. van den Berg A., Draaijers T. G., Kok K., Timmer T., Van der Veen A. Y., Veldhuis P. M., de Leij L., Gerhartz C. D., Naylor S. L., Smith D. I., Buys C. H. Normal FHIT transcripts in renal cell cancer- and lung cancer-derived cell lines, including a cell line with a homozygous deletion in the FRA3B region. Genes Chromosomes Cancer, 19: 220-227, 1997.[Medline]



This article has been cited by other articles:


Home page
 Cancer Epidemiol. Biomarkers Prev.Home page
P. Bavi, Z. Jehan, V. Atizado, H. Al-Dossari, F. Al-Dayel, A. Tulbah, S. S. Amr, S. S. Sheikh, A. Ezzat, H. El-Solh, et al.
Prevalence of fragile histidine triad expression in tumors from saudi arabia: a tissue microarray analysis.
Cancer Epidemiol. Biomarkers Prev., September 1, 2006; 15(9): 1708 - 1718.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
F. D'Agostini, A. Izzotti, R. Balansky, N. Zanesi, C. M. Croce, and S. De Flora
Early loss of fhit in the respiratory tract of rodents exposed to environmental cigarette smoke.
Cancer Res., April 1, 2006; 66(7): 3936 - 3941.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
K. Mimori, H. Ishii, H. Nagahara, T. Sudo, K. Yamashita, H. Inoue, G. F. Barnard, and M. Mori
FHIT Is Up-Regulated by Inflammatory Stimuli and Inhibits Prostaglandin E2-Mediated Cancer Progression.
Cancer Res., March 1, 2006; 66(5): 2683 - 2690.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
U.-P. Rohr, N. Rehfeld, H. Geddert, L. Pflugfelder, I. Bruns, J. Neukirch, A. Rohrbeck, H. J. Grote, U. Steidl, R. Fenk, et al.
Prognostic Relevance of Fragile Histidine Triad Protein Expression in Patients with Small Cell Lung Cancer
Clin. Cancer Res., January 1, 2005; 11(1): 180 - 185.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
C. Lu, J.-C. Soria, X. Tang, X.-C. Xu, L. Wang, L. Mao, R. Lotan, B. Kemp, B. N. Bekele, L. Feng, et al.
Prognostic Factors in Resected Stage I Non-Small-Cell Lung Cancer: A Multivariate Analysis of Six Molecular Markers
J. Clin. Oncol., November 15, 2004; 22(22): 4575 - 4583.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
J. S. Kim, H. Kim, Y. M. Shim, J. Han, J. Park, and D.-H. Kim
Aberrant methylation of the FHIT gene in chronic smokers with early stage squamous cell carcinoma of the lung
Carcinogenesis, November 1, 2004; 25(11): 2165 - 2171.
[Abstract] [Full Text] [PDF]

Home page
 Eur Respir JHome page
C. Mascaux, B. Martin, J.M. Verdebout, A.P. Meert, V. Ninane, and J.P. Sculier
Fragile histidine triad protein expression in nonsmall cell lung cancer and correlation with Ki-67 and with p53
Eur. Respir. J., May 1, 2003; 21(5): 753 - 758.
[Abstract] [Full Text] [PDF]

Home page
 Pharmacol. Rev.Home page
R. Danesi, F. De Braud, S. Fogli, T. M. De Pas, A. Di Paolo, G. Curigliano, and M. Del Tacca
Pharmacogenetics of Anticancer Drug Sensitivity in Non-Small Cell Lung Cancer
Pharmacol. Rev., March 1, 2003; 55(1): 57 - 103.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
Y. S. Chang, L. Wang, D. Liu, L. Mao, W. K. Hong, F. R. Khuri, and H.-Y. Lee
Correlation between Insulin-like Growth Factor-binding Protein-3 Promoter Methylation and Prognosis of Patients with Stage I Non-Small Cell Lung Cancer
Clin. Cancer Res., December 1, 2002; 8(12): 3669 - 3675.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
K. Uematsu, A. Yoshimura, A. Gemma, H. Mochimaru, Y. Hosoya, S. Kunugi, K. Matsuda, M. Seike, F. Kurimoto, K. Takenaka, et al.
Aberrations in the Fragile Histidine Triad (FHIT) Gene in Idiopathic Pulmonary Fibrosis
Cancer Res., December 1, 2001; 61(23): 8527 - 8533.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
T. Tsujiuchi, Y. Sasaki, N. Murata, M. Tsutsumi, Y. Konishi, and D. Nakae
FHIT alterations in lung adenocarcinomas induced by N-nitrosobis(2-hydroxypropyl)amine in rats
Carcinogenesis, December 1, 2001; 22(12): 2017 - 2022.
[Abstract] [Full Text] [PDF]

Home page
 JAMAHome page
H. Ishii, K. R. Dumon, A. Vecchione, L. Y. Y. Fong, R. Baffa, K. Huebner, and C. M. Croce
Potential Cancer Therapy With the Fragile Histidine Triad Gene: Review of the Preclinical Studies
JAMA, November 21, 2001; 286(19): 2441 - 2449.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
S. Zöchbauer-Müller, K. M. Fong, A. Maitra, S. Lam, J. Geradts, R. Ashfaq, A. K. Virmani, S. Milchgrub, A. F. Gazdar, and J. D. Minna
5' CpG Island Methylation of the FHIT Gene Is Correlated with Loss of Gene Expression in Lung and Breast Cancer
Cancer Res., May 1, 2001; 61(9): 3581 - 3585.
[Abstract] [Full Text]

Home page
 Cancer Res.Home page
J. Brognard, A. S. Clark, Y. Ni, and P. A. Dennis
Akt/Protein Kinase B Is Constitutively Active in Non-Small Cell Lung Cancer Cells and Promotes Cellular Survival and Resistance to Chemotherapy and Radiation
Cancer Res., May 1, 2001; 61(10): 3986 - 3997.
[Abstract] [Full Text]

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 Tseng, J. E.
Right arrow Articles by Mao, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tseng, J. E.
Right arrow Articles by Mao, L.

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 Meeting Abstracts Online