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Clinical Cancer Research Vol. 8, 2292-2297, July 2002
© 2002 American Association for Cancer Research

Molecular Oncology, Markers, Clinical Correlates

Differential Expression of FEZ1/LZTS1 Gene in Lung Cancers and Their Cell Cultures1

Shinichi Toyooka, Yasuro Fukuyama, Ignacio I. Wistuba, Melvyn S. Tockman, John D. Minna and Adi F. Gazdar2

Hamon Center for Therapeutic Oncology Research [S. T., Y. F., J. D. M., A. F. G.], and Departments of Pathology [A. F. G.], Internal Medicine [J. D. M.], and Pharmacology [J. D. M.], University of Texas Southwestern Medical Center, Dallas, Texas 75390-8593; Department of Pathology, Pontificia Universidad Catolica de Chile, Santiago, Chile [I. I. W.]; and Molecular Screening Laboratory, H. Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, Florida 33612-9497 [M. S. T.]


   ABSTRACT
Top
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Purpose: The FEZ1/LZTS1 (FEZ1) gene, located on chromosome 8p22 (8p22), was identified recently as a candidate tumor suppressor gene. Because loss of heterozygosity at 8p21–22 is a frequent event in lung cancers, we studied FEZ1 alteration in short-term cultures of resected lung cancer tumors and cell lines.

Experimental Design: We examined FEZ1 expression in 17 non-small cell lung cancer (NSCLC), 19 small cell lung cancer (SCLC) cell lines, and 6 pairs of short-term cultures of resected NSCLCs and accompanying nonmalignant bronchial cells (NBECs) by reverse transcription-PCR and Western blotting. To investigate the mechanism for silencing, cells were cultured with 5-aza-2'-deoxycytidine or trichostatin A. We screened for genomic mutations by PCR-single-strand conformational polymorphism.

Results: Thirteen of 17 NSCLC (76%) and 3 of 19 SCLC (16%) of cell lines showed absent expression (P = 0.001). Of the paired NSCLC-NBEC cultures, 3 of 6 showed loss of expression in tumor cell cultures. In the cell lines retaining expression, the amplicon products in SCLCs were more intense than those of NSCLCs and NBECs. Expression of FEZ1 was not restored by 5-aza-2'-deoxycytidine and trichostatin A. Although FEZ1 expression was moderately correlated with loss of heterozygosity of specific microsatellite makers at 8p21–22 in NSCLC cell lines, it was strongly correlated to D8S261 and LPL loci in SCLC cell lines. No mutation was found within cording region of FEZ1 by PCR-single-strand conformational polymorphism.

Conclusions: We found differential FEZ1 expression in NSCLC and SCLC cell lines, and the absent expression in 3 of 6 short-term cultures of NSCLC tumors. FEZ1 may be related to tumorigenesis of lung cancer.


   INTRODUCTION
Top
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Lung cancer is the most common cause of cancer deaths in the United States (1) and on clinicopathological grounds is divided into two major types, NSCLCs and SCLCs. The molecular genetic changes in these two types of lung cancer are very different, including specific patterns of allelic loss (2, 3, 4, 5) . Frequent allelic loss at the short arm of chromosome 8p21–23 (8p21–23) region has been reported in lung cancers (2 , 3 , 6, 7, 8, 9) . In microdissected lung cancers, we found LOH3 in 86% of SCLCs, 100% of squamous cell carcinomas, and 81% of adenocarcinomas (7) . Furthermore, deletions commence early during the multistage development at the hyperplasia/metaplasia stage in NSCLC tumor patients and in smokers without cancer (7) . The deletions may persist for several decades after smoking cessation (7) .

Allelic losses on the 8p have also been reported as a frequent event in several kinds of cancer including prostate, colon, breast, head and neck, urinary bladder, hepatocellular, and cholangiocarcinomas (10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21) . These findings suggest the presence of TSGs in this region. Functional evidence for this hypothesis was provided by chromosome transfer and microcell fusion experiments (22, 23, 24, 25) .

At least two major candidate TSGs have been identified on 8p (26 , 27) . One of these, the FEZ1/LZTS1 (FEZ1) gene located on 8p22 encodes a Mr 67,000 leucine-zipper protein (27) . Genomic mutation and loss of expression of FEZ1 was reported in some solid tumors including esophagus, prostate, and gastric cancers (27 , 28) . In this study, we examined the expression and mutation status of FEZ1 gene in NSCLC and SCLC cell lines, and also primary cultures of NSCLC tumors and corresponding NBECs.


   MATERIALS AND METHODS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Lung Cancer Cell Lines.
Thirty six lung cancer cell lines (17 NSCLC and 19 SCLC cell lines) and corresponding B lymphoblastoid lines (n = 30) were established by us (29) . The 17 NSCLC cell lines consisted of 13 adenocarcinomas, 2 squamous cell carcinomas, and 2 large cell carcinomas. Most NSCLC cell lines were derived from primary tumors, and most SCLC cell lines were from metastases. Cells cultures were grown in RPMI 1640 (Life Technologies, Inc., Rockville, MD) supplemented with 5% fetal bovine serum and incubated in 5% CO2 at 37°C.

Primary Culture of Resected Lung Cancer and NBECs.
Six NSCLC primary cells (2 cases each of adenocarcinomas, 2 squamous cell carcinomas, and 2 large carcinomas) and its corresponding NBECs were cultured. Primary cultures of resected nonmalignant lung tissue were selected for culture and placed in trypsin in the cold room for 22–24 h. The next day, the specimen was removed from the cold room and 1 ml of medium was added. The medium was MCDB1532+, consisting of MCDB 153 basal medium (Sigma Chemical Company, St. Louis, MO) supplemented with growth factors in a collagen-coated dish. Every day, fresh medium was added to the dish. For primary lung tumor cell culture, small tumor pieces were enzymatically disassociated and cultured in MCDB153+ medium. All of the culture pairs have been immunophenotyped with vimentin, cytokeratin, and thyroid transcription factor. Early passages from all of the lines were preserved in liquid nitrogen until tested.

RT-PCR Assay.
The RT-PCR assay was used to examine FEZ1 mRNA expression. Total RNA was extracted from the samples (6 NSCLC primary cells and its corresponding NBECs, 17 NSCLC, and 19 SCLC cell lines) with Trizol (Life Technologies, Inc.) following the manufacturer’s instructions. Reverse transcription reaction was performed on 2 µg of total RNA with the SuperScript II First-Strand Synthesis using oligodeoxythymidylic acid primer System (Life Technologies, Inc.), and aliquots of the reaction mixture were used for the subsequent PCR amplification. The forward PCR amplification primer was 5'-GAGCCTCATGAAGGAGCAGG-3' targeted the end of exon 2, and the reverse primer 5'-CAGGTCCTGGGTCCTCAGGT-3' the beginning of exon 3 of FEZ1 gene, and confirmed that genomic DNA was not amplified with these primers. PCR amplification was carried out in a reaction volume of 25 µl for 12 min at 95°C for initial denaturation, followed by 33 cycles of 94°C for 20 s, 60°C for 30 s, and 72°C for 30 s. The housekeeping gene GAPDH was used as an internal control to confirm the success of the reverse transcription reaction. The primers for GAPDH amplification were followed: forward primer, 5'-ACAGTCCATGCCATCACTGCC-3' and reverse primer, 5'-GCCTGCTTCACCACCTTCTTG-3'. PCR products were analyzed on 2% agarose gels.

5-Aza-CdR and TSA Treatment.
Six lung cancer cell lines (4 cases of NSCLC and 2 cases of SCLC) with loss of FEZ1 expression were incubated in culture medium with 5-Aza-CdR and TSA each (30) . Drug treatment was accomplished by adding reagents to the culture medium to final concentrations as follows: (a) 5-Aza-CdR, 2 µg/ml; and (b) TSA, 150 and 300 nM. Cells were treated with 5-Aza-CdR for 5 days and TSA for 24–48 h. Medium were changed every 48 h for 5-Aza-CdR and every 24 h for TSA.

Western Blot Analysis.
A sample of protein (30 µg) from the cell lysates were separated by SDS-PAGE in 10% polyacrylamide gels and transferred to nitrocellulose membranes. The membranes were incubated first with primary antibody to Fez1 protein kindly given by Drs. Carlo Croce and Hideshi Ishii (Kimmel Cancer Center, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA) and then with rabbit polyclonal antibody as secondary antibody coupled to horseradish peroxidase, after which the membranes were developed by SuperSignal West Pico Chemiluminescent Substrate (PIERCE, Rockford, IL).

DNA Extraction and PCR-SSCP Assay.
Genomic DNA was isolated from all of the cultured cells by SDS/proteinase K (Life Technologies, Inc.) digestion, phenol-chloroform extraction, and ethanol precipitation (31) . PCR-SSCP assay was performed for the cording region of FEZ1 gene. The primer information used for PCR-SSCP is summarized in Table 1Citation . PCR amplification was carried out 12 min for 95°C for initial denaturation, followed by 35 cycles of 94°C for 20 s, 60–64°C for 40 s, and 72°C for 40 s. The PCR products were then denatured, loaded on 6% polyacrylamide gels with and without 5% (vol/vol) glycerol, electrophoresed, and exposed to X-ray film.


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  		Table 1 Primer sequences for PCR-SSCP

 
Data Analysis.
The frequencies of FEZ1 expression in NSCLCs and SCLCs were compared using {chi}2 tests. Probability values of P < 0.05 were regarded as statistically significant. The statistical test was two-sided.


   RESULTS
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
We analyzed FEZ1 expression of mRNA in 17 NSCLC and 19 SCLC cell lines by RT-PCR (Table 2)Citation . Thirteen of 17 (76%) NSCLC cell lines lacked FEZ1 expression. By contrast, only 3 of 19 (16%) SCLC cell lines lacked FEZ1 expression. The difference of EFZ1 expression status between NSCLCs and SCLCs was significant (P = 0.001). There was no difference in expression status between adenocarcinoma, squamous cell carcinoma, and large cell carcinoma cell lines. A total of 18 cell lines (9 NSCLCs and 9 SCLCs) were tested for protein expression by Western blot, and the concordance with the RT-PCR expression was 100% (Table 2)Citation . Representative samples are shown in Figs. 1Citation and 2Citation . Of interest, in positive cell lines, expression of FEZ1 by both RT-PCR and Western blot was stronger in SCLC cell lines than in NSCLC cell lines (Figs. 1Citation and 2Citation ). Expression of FEZ1 was not restored by neither 5-Aza-CdR nor TSA treatment in 6 cell lines of which FEZ1 expression was negative.


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  		Table 2 FEZ1 mRNA expression in NSCLC and SCLC cell lines

 


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  		Fig. 1. Representative examples of RT-PCR assay for FEZ1 RNA expression in NSCLC and SCLC cell lines. Expression of the housekeeping gene GAPDH was used as a control for RNA integrity. PC, positive control; NC, negative control (genomic DNA); [NS], NSCLC; [S], SCLC.

 


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  		Fig. 2. Representative figure of Western bolt analysis for Fez1 in NSCLC and SCLC cell lines. Expression of the housekeeping gene Actin was used as a control for protein loading. [NS], NSCLC; [S], SCLC.

 
Because of stromal cell contamination, we could not test tumors directly for loss of FEZ1 expression. Thus, we chose to test short-term cultures of NSCLC tumors and corresponding NBECs. These cells were cultured under conditions that suppressed stromal cell proliferation, and they consisted entirely of epithelial cells. All 6 of the NBEC cultures and 3 (50%) of the NSCLC cultures demonstrated FEZ1 expression by RT-PCR (Fig. 3Citation ; Table 3Citation ). The number of cultured cells available was insufficient to perform Western blot study.



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  		Fig. 3. RT-PCR assay for FEZ1 and GAPDH in 6 NSCLCs and their corresponding NBEC primary cultures. Expression of the housekeeping gene GAPDH was used as a control for RNA integrity. PC, positive control; NC, negative control (genomic DNA); T, NSCLC cell culture sample; N, corresponding nonmalignant epithelial cell culture sample.

 

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  		Table 3 FEZ1 mRNA expression in primary NSCLC and corresponding bronchial epithelial cell cultures

 
We compared the presence or absence of LOH at 8p21–22 using three polymorphic markers with FEZ1 expression in 17 NSCLC and 13 SCLC cell lines (Table 2)Citation . Except for 2 NSCLC cell lines, all were informative for at least one marker. LOH at one or more markers was present in 9 of 15 (60%) informative NSCLC cases and in 5 of 13 (31%) informative SCLC cases. The concordances between LOH and FEZ1expression status were summarized in Table 4Citation . The FEZ1 expression was moderately correlated with LOH of tested microsatellite makers at 8p21–22 in NSCLC cell lines, it was strongly correlated to D8S261 and LPL loci in SCLC cell lines (both concordances were 82%).


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  		Table 4 Concordances between FEZ1 expression and 8p21–22 polymorphic markers in NSCLC and SCLC cell lines

Data of all the informative cell lines are presented. Loss of FEZ1 expression was correlated with allelic loss at three informative markers near the FEZ1 gene at 8p21–22.

 
We examined the genomic mutation by PCR-SSCP using eight sets of primer covering cording region of FEZ1 gene. However, no evidence of genomic mutation was detected by this technique.


   DISCUSSION
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
FEZ1 was identified recently as a candidate TSG at 8p22, and absence or reduction of FEZ1 expression and occasional genomic mutation were reported in several kinds of cancers (27) . Introduction of FEZ1 into Fez1-negative cells resulted in inhibition of cell growth, suggesting that FEZ1 is a candidate TSG. Fez1 is associated with microtubule components and interacts with p34 (cdc2) at late S-G2/M stage in vivo (32) .

Our data demonstrate a highly significant difference (P = 0.001) between the rates of FEZ1 inactivation in NSCLC (76%) and SCLC (16%) cell lines. There was complete concordance between RNA expression (as detected by RT-PCR) and protein expression (as detected by Western blot). These findings are in accord with previous reports of more frequent allelic loss at the gene location (8p22) in NSCLC carcinomas and cell lines compared with SCLC cell lines (7) . Expression data correlated with allelic loss, particularly with LOH at marker D8S261. Examination of a limited number of short-term cultures of paired NSCLC tumors and their corresponding bronchial epithelial cultures demonstrated expression in all of the NBEC cultures but loss of expression in 50% of the paired tumors. This illustrates that the loss of FEZ1 was likely sustained during neoplastic transformation and is not an artifact of long-term cell culture. Expression in SCLC cell lines was relatively strong compared with NSCLC cell lines, tumors, and nonmalignant bronchial epithelium. Whereas the origin of SCLC is controversial, one possibility is that they arise from bronchial cells programmed to become neuroendocrine cells and that FEZ1 expression in neuroendocrine cells is greater than in the other epithelial cells. Of interest, FEZ1 is strongly expressed in brain tissue (27) .

The mechanism of down-regulation of FEZ1 expression in human cancers has not been demonstrated convincingly. Mutations are relatively rare, and we could find no evidence of mutations in expression negative lung cancer cell lines. Recently aberrant methylation of CpG islands in the promoter region has become established as an important epigenetic mechanism for gene silencing (33) . We reported that the profile of aberrant methylation was different between NSCLCs and SCLCs (34) . Vecchione, et al. (28) reported that hypermethylation of 5' region of FEZ1 (1 kb including exon 2) was present in gastric cancer cell lines and also peripheral leukocyte DNA; however, the status of methylation of the 5' region of FEZ1 was not correlated with the level of expression. Because the expression of FEZ1 was not restored by the 5-Aza-CdR treatment in our study, aberrant methylation may not be related to silencing of FEZ1 expression in both NSCLCs and SCLCs. Like other mechanisms for controlling gene expression, histone deacetylation causes silencing of gene expression (35) . However, failure of restoration of FEZ1 expression by treatment with the histone deacetylation inhibitor TSA indicated that histone deacetylation was not the cause of silencing of FEZ1 expression. Thus, the mechanism of FEZ1 gene silencing in lung and other tumors remains unknown.

Our findings indicate that FEZ1 is inactivated frequently in NSCLC tumors and cell lines, and that inactivation is associated with frequent allelic loss at 8p21–22. These findings support a role for FEZ1 as a TSG in NSCLCs. Of interest, other oncogenes and TSGs demonstrate striking differences between the two major forms of lung cancer, including RAS, MYC amplification, and inactivation of the p16 and caspase 8 genes (36, 37, 38, 39) . These studies support the concept that these forms of lung cancer may arise via different pathogenetic pathways.


   ACKNOWLEDGMENTS
 
We thank Drs. Hideshi Ishii and Carlo M. Croce (Kimmel Cancer Center, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA) for the kind gift of Fez1 antibody.


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

1 Supported by grants from the University of Texas Specialized Program of Research Excellence in Lung Cancer (P50CA70907) and the Early Detection Research Network (5U01CA8497102), National Cancer Institute, Bethesda, MD. Back

2 To whom requests for reprints should be addressed, at Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, 6000 Harry Hines Boulevard, Dallas, Texas 75390-8593. Phone: (214) 648-4921; Fax: (214) 648-4940; E-mail: Adi.gazdar{at}UTsouthwestern.edu Back

3 The abbreviations used are: LOH, loss of heterozygosity; SCLC, small cell lung cancer; NSCLC, non-small cell lung cancer; TSG, tumor suppressor gene; RT-PCR, reverse transcription-PCR; NBEC, nonmalignant bronchial cell; 5-Aza-CdR, 5-aza-2'-deoxycytidine; TSA, trichostatin A; SSCP, single-strand conformational polymorphism. Back

Received 2/18/02; revised 4/15/02; accepted 4/15/02.


   REFERENCES
Top
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. Greenlee R. T., Murray T., Bolden S., Wingo P. A. Cancer statistics, 2000. CA Cancer J. Clin., 50: 7-33, 2000.[Abstract]
  2. Virmani A. K., Fong K. M., Kodagoda D., McIntire D., Hung J., Tonk V., Minna J. D., Gazdar A. F. Allelotyping demonstrates common and distinct patterns of chromosomal loss in human lung cancer types. Genes Chromosomes Cancer, 21: 308-319, 1998.[CrossRef][Medline]
  3. Girard L., Zochbauer-Muller S., Virmani A. K., Gazdar A. F., Minna J. D. Genome-wide allelotyping of lung cancer identifies new regions of allelic loss, differences between small cell lung cancer and non-small cell lung cancer, and loci clustering. Cancer Res., 60: 4894-906, 2000.[Abstract/Free Full Text]
  4. Sekido Y., Fong K. M., Minna J. D. Progress in understanding the molecular pathogenesis of human lung cancer. Biochim. Biophys. Acta, 1378: F21-F59, 1998.[Medline]
  5. Wistuba I. I., Barry J., Behrens C., Maitra A., Shivapurkar N., Milchgrub S., Mackay B., Minna J. D., Gazdar A. F. Molecular changes in the bronchial epithelium of patients with small cell lung cancer. Clin. Cancer Res., 6: 2604-2610, 2000.[Abstract/Free Full Text]
  6. Lerebours F., Olschwang S., Thuille B., Schmitz A., Fouchet P., Buecher B., Martinet N., Galateau F., Thomas G. Fine deletion mapping of chromosome 8p in non-small-cell lung carcinoma. Int. J. Cancer, 81: 854-858, 1999.[CrossRef][Medline]
  7. Wistuba I. I., Behrens C., Virmani A. K., Milchgrub S., Syed S., Lam S., Mackay B., Minna J. D., Gazdar A. F. Allelic losses at chromosome 8p21–23 are early and frequent events in the pathogenesis of lung cancer. Cancer Res., 59: 1973-1979, 1999.[Abstract/Free Full Text]
  8. Emi M., Fujiwara Y., Ohata H., Tsuda H., Hirohashi S., Koike M., Miyaki M., Monden M., Nakamura Y. Allelic loss at chromosome band 8p21.3-p22 is associated with progression of hepatocellular carcinoma. Genes Chromosomes Cancer, 7: 152-157, 1993.[Medline]
  9. Ohata H., Emi M., Fujiwara Y., Higashino K., Nakagawa K., Futagami R., Tsuchiya E., Nakamura Y. Deletion mapping of the short arm of chromosome 8 in non-small cell lung carcinoma. Genes Chromosomes Cancer, 7: 85-88, 1993.[Medline]
  10. Kagan J., Stein J., Babaian R. J., Joe Y. S., Pisters L. L., Glassman A. B., von Eschenbach A. C., Troncoso P. Homozygous deletions at 8p22 and 8p21 in prostate cancer implicate these regions as the sites for candidate tumor suppressor genes. Oncogene, 11: 2121-2126, 1995.[Medline]
  11. Bova G. S., Carter B. S., Bussemakers M. J., Emi M., Fujiwara Y., Kyprianou N., Jacobs S. C., Robinson J. C., Epstein J. I., Walsh P. C., et al Homozygous deletion and frequent allelic loss of chromosome 8p22 loci in human prostate cancer. Cancer Res., 53: 3869-3873, 1993.[Abstract/Free Full Text]
  12. Kerangueven F., Essioux L., Dib A., Noguchi T., Allione F., Geneix J., Longy M., Lidereau R., Eisinger F., Pebusque M. J., et al Loss of heterozygosity and linkage analysis in breast carcinoma: indication for a putative third susceptibility gene on the short arm of chromosome 8. Oncogene, 10: 1023-1026, 1995.[Medline]
  13. Yaremko M. L., Wasylyshyn M. L., Paulus K. L., Michelassi F., Westbrook C. A. Deletion mapping reveals two regions of chromosome 8 allele loss in colorectal carcinomas. Genes Chromosomes Cancer, 10: 1-6, 1994.[Medline]
  14. Yaremko M. L., Recant W. M., Westbrook C. A. Loss of heterozygosity from the short arm of chromosome 8 is an early event in breast cancers. Genes Chromosomes Cancer, 13: 186-191, 1995.[Medline]
  15. Emi M., Fujiwara Y., Nakajima T., Tsuchiya E., Tsuda H., Hirohashi S., Maeda Y., Tsuruta K., Miyaki M., Nakamura Y. Frequent loss of heterozygosity for loci on chromosome 8p in hepatocellular carcinoma, colorectal cancer, and lung cancer. Cancer Res., 52: 5368-5372, 1992.[Abstract/Free Full Text]
  16. Fujiwara Y., Emi M., Ohata H., Kato Y., Nakajima T., Mori T., Nakamura Y. Evidence for the presence of two tumor suppressor genes on chromosome 8p for colorectal carcinoma. Cancer Res., 53: 1172-1174, 1993.[Abstract/Free Full Text]
  17. Knowles M. A., Shaw M. E., Proctor A. J. Deletion mapping of chromosome 8 in cancers of the urinary bladder using restriction fragment length polymorphisms and microsatellite polymorphisms. Oncogene, 8: 1357-1364, 1993.[Medline]
  18. MacGrogan D., Levy A., Bostwick D., Wagner M., Wells D., Bookstein R. Loss of chromosome arm 8p loci in prostate cancer: mapping by quantitative allelic imbalance. Genes Chromosomes Cancer, 10: 151-159, 1994.[Medline]
  19. Ichii S., Takeda S., Horii A., Nakatsuru S., Miyoshi Y., Emi M., Fujiwara Y., Koyama K., Furuyama J., Utsunomiya J., et al Detailed analysis of genetic alterations in colorectal tumors from patients with and without familial adenomatous polyposis (FAP). Oncogene, 8: 2399-405, 1993.[Medline]
  20. Yaremko M. L., Kutza C., Lyzak J., Mick R., Recant W. M., Westbrook C. A. Loss of heterozygosity from the short arm of chromosome 8 is associated with invasive behavior in breast cancer. Genes Chromosomes Cancer, 16: 189-195, 1996.[CrossRef][Medline]
  21. Kawaki J., Miyazaki M., Ito H., Nakagawa K., Shimizu H., Yoshidome H., Uzawa K., Tanzawa H., Nakajima N. Allelic loss in human intrahepatic cholangiocarcinoma: correlation between chromosome 8p22 and tumor progression. Int. J. Cancer, 88: 228-231, 2000.[CrossRef][Medline]
  22. Gustafson C. E., Wilson P. J., Lukeis R., Baker E., Woollatt E., Annab L., Hawke L., Barrett J. C., Chenevix-Trench G. Functional evidence for a colorectal cancer tumor suppressor gene at chromosome 8p22–23 by monochromosome transfer. Cancer Res., 56: 5238-5245, 1996.[Abstract/Free Full Text]
  23. Ichikawa T., Nihei N., Suzuki H., Oshimura M., Emi M., Nakamura Y., Hayata I., Isaacs J. T., Shimazaki J. Suppression of metastasis of rat prostatic cancer by introducing human chromosome 8. Cancer Res., 54: 2299-302, 1994.[Abstract/Free Full Text]
  24. Kuramochi H., Ichikawa T., Nihei N., Kawana Y., Suzuki H., Schalken J. A., Takeichi M., Nagafuchi A., Ito H., Shimazaki J. Suppression of invasive ability of highly metastatic rat prostate cancer by introduction of human chromosome 8. Prostate, 31: 14-20, 1997.[CrossRef][Medline]
  25. Nihei N., Ichikawa T., Kawana Y., Kuramochi H., Kugoh H., Oshimura M., Hayata I., Shimazaki J., Ito H. Mapping of metastasis suppressor gene(s) for rat prostate cancer on the short arm of human chromosome 8 by irradiated microcell-mediated chromosome transfer. Genes Chromosomes Cancer, 17: 260-268, 1996.[CrossRef][Medline]
  26. Fujiwara Y., Ohata H., Kuroki T., Koyama K., Tsuchiya E., Monden M., Nakamura Y. Isolation of a candidate tumor suppressor gene on chromosome 8p21.3-p22 that is homologous to an extracellular domain of the PDGF receptor ß gene. Oncogene, 10: 891-895, 1995.[Medline]
  27. Ishii H., Baffa R., Numata S. I., Murakumo Y., Rattan S., Inoue H., Mori M., Fidanza V., Alder H., Croce C. M. The FEZ1 gene at chromosome 8p22 encodes a leucine-zipper protein, and its expression is altered in multiple human tumors. Proc. Natl. Acad. Sci. USA, 96: 3928-3933, 1999.[Abstract/Free Full Text]
  28. Vecchione A., Ishii H., Shiao Y. H., Trapasso F., Rugge M., Tamburrino J. F., Murakumo Y., Alder H., Croce C. M., Baffa R. Fez1/lzts1 alterations in gastric carcinoma. Clin. Cancer Res., 7: 1546-1552, 2001.[Abstract/Free Full Text]
  29. Gazdar A. F., Kurvari V., Virmani A., Gollahon L., Sakaguchi M., Westerfield M., Kodagoda D., Stasny V., Cunningham H. T., Wistuba I. I., Tomlinson G., Tonk V., Ashfaq R., Leitch A. M., Minna J. D., Shay J. W. Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer. Int. J. Cancer, 78: 766-774, 1998.[CrossRef][Medline]
  30. Toyooka, S., Carbone, M., Toyooka, K. O., Bocchetta, M., Shivapurkar, N., Minna, J. D., and Gazdar, A. F. Progressive aberrant methylation of the RASSF1A gene in simian virus 40 infected human mesothelial cells, Oncogene, in press, 2002.
  31. Herrmann B. G., Frischauf A. M. Isolation of genomic DNA. Methods Enzymol., 152: 180-183, 1987.[Medline]
  32. Ishii H., Vecchione A., Murakumo Y., Baldassarre G., Numata S., Trapasso F., Alder H., Baffa R., Croce C. M. FEZ1/LZTS1 gene at 8p22 suppresses cancer cell growth and regulates mitosis, Proc. Natl. Acad. Sci. USA, 98: 10374-10379, 2001.[Abstract/Free Full Text]
  33. Herman J. G., Merlo A., Mao L., Lapidus R. G., Issa J. P., Davidson N. E., Sidransky D., Baylin S. B. Inactivation of the CDKN2/p16/MTS1 gene is frequently associated with aberrant DNA methylation in all common human cancers. Cancer Res., 55: 4525-4530, 1995.[Abstract/Free Full Text]
  34. Toyooka S., Toyooka O. K., Maruyama R., Virmani A. K., Girard L., Miyajima K., Harada K., Ariyoshi Y., Takahashi T., Sugio K., Brambilla E., Gilcrease M., Minna J. D., Gazdar A. F. DNA methylation profiles of lung tumors. Mol. Cancer Ther., 1: 61-67, 2001.[Abstract/Free Full Text]
  35. Cameron E. E., Bachman K. E., Myohanen S., Herman J. G., Baylin S. B. Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer. Nat. Genet., 21: 103-107, 1999.[CrossRef][Medline]
  36. Kiefer P. E., Wegmann B., Bacher M., Erbil C., Heidtmann H., Havemann K. Different pattern of expression of cellular oncogenes in human non- small-cell lung cancer cell lines. J. Cancer Res. Clin. Oncol., 116: 29-37, 1990.[CrossRef][Medline]
  37. Mitsudomi T., Viallet J., Mulshine J. L., Linnoila R. I., Minna J. D., Gazdar A. F. Mutations of ras genes distinguish a subset of non-small-cell lung cancer cell lines from small-cell lung cancer cell lines. Oncogene, 6: 1353-1362, 1991.[Medline]
  38. Kelley M. J., Nakagawa K., Steinberg S. M., Mulshine J. L., Kamb A., Johnson B. E. Differential inactivation of CDKN2 and Rb protein in non-small-cell and small-cell lung cancer cell lines. J. Natl. Cancer Inst., 87: 756-761, 1995.[Abstract/Free Full Text]
  39. Shivapurkar N., Toyooka S., Eby M. T., Huang C. X., Sathyanarayana U. G., Cunningham H. T., Reddy J. L., Brambilla E., Takahashi T., Minna J. D., Chaudhary P. M., Gazdar A. F. Differential inactivation of caspase-8 in lung cancers. Cancer Biology and Therapy, 1: 65-69, 2002.



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