| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Molecular Biology and Genetics |
Dipartimento di Patologia [C. S., A. B., S. O., G. Z., A. S.] and Dipartimento di Scienze Chirurgiche [P. P.], Università di Verona, I-37134, Verona, Italy; and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107 [K. H.]
ABSTRACT
We examined 2 normal pancreata, 21 primary pancreatic ductal cancers, and 19 pancreatic cancer cell lines for Fhit expression and FHIT gene status. The normal pancreas expressed Fhit protein in the cytoplasm of ductular cells, whereas interlobular and larger ducts, acini, and insulae of Langerhans were negative. Fhit protein was detected by immunoblot assay in 11 pancreatic cancer cell lines; of the 8 cell lines lacking Fhit protein, 7 lacked FHIT mRNA and 1 showed an abnormally sized transcript. DNA from five of these eight cell lines showed homozygous loss of FHIT exon 5. In 8 of the 21 primary cancers, Fhit expression was detected by immunohistochemistry. Reverse transcription-PCR analysis of 6 of the 13 cases lacking Fhit showed normal-sized FHIT product in 3 cases and a mixture of normal and abnormal products in the other 3. Sequencing showed that abnormal bands were missing variable numbers of exons. Loss of microsatellite DNA markers internal to the FHIT gene was observed in 10 of 13 primary cancers lacking Fhit protein (homozygous in two cases) and in only 1 of the 8 cancers expressing Fhit protein. In nine primary cancers, four expressing and five lacking Fhit protein, it was possible to obtain pure cancer DNA by microdissection. Three of the five microdissected cases lacking Fhit protein exhibited homozygous deletion of FHIT exon 5. In conclusion, the lack of Fhit protein in pancreatic cancers correlated with absence or alteration of FHIT mRNA and was often associated with FHIT gene anomalies.
INTRODUCTION
Chromosome 3p is among the chromosomes that show relatively frequent karyotype anomalies and allelic losses in pancreatic cancer (1, 2, 3, 4)
. In the course of allelotyping studies of cryostat-enriched primary pancreatic cancers, we found allelic losses at chromosome 3p14.2 in 
60% of cases and rarely on chromosome 3 outside of this region, paralleling the findings of Shridhar et al. (5)
. The fragile histidine triad gene (FHIT) is located at this site and has been proposed as a candidate tumor suppressor gene, due to its frequent alteration in different cancer types (6, 7, 8)
. It consists of 10 exons distributed over a genomic region of >1 Mb, which also contains the FRA3B fragile site, the most inducible of the common aphidicolin-inducible fragile sites. The 1.1-kb FHIT transcript encodes a protein with in vitro dinucleoside 5',5''''-P1,P3- triphosphate hydrolase activity, but its in vivo function is as yet unknown (9)
.
A considerable amount of data has accumulated over the last few years on FHIT anomalies in different cancer types (6 , 7 , 10, 11, 12, 13, 14, 15) . The anomalies included the frequent presence of abnormal FHIT transcripts detected by RT-PCR3 and the absence of Fhit protein expression, often associated with FHIT gene alterations (for reviews see Refs. 8 , 16 , and 17 ). These mainly included deletions of different arrays of FHIT exons. Lung, stomach, kidney, and cervical cancers derive from normal epithelial tissues that are strongly positive for Fhit expression, yet >50% of cancers from each of these organs has lost Fhit expression (12 , 18, 19, 20) . The available data on FHIT in pancreatic cancer are limited to the analysis of LOH in primary tumors (5) and the detailed analysis of 17 cancer cell lines (5 , 21 , 22) . Among the 13 cell lines analyzed by Simon et al. (21) , 4 had wild-type FHIT transcripts, whereas 9 exhibited absent or abnormal mRNAs. Four additional pancreatic cancer cell lines were reported as showing apparent integrity of FHIT transcripts (5 , 22) . Homozygous losses of various 3p14.2 microsatellite markers or FHIT exons were also found (21) . No data are available on FHIT gene status and protein expression in primary pancreatic cancers or normal pancreas.
Here, we examined two normal pancreata and a series of pancreatic ductal carcinomas for Fhit protein and mRNA expression as well as for the presence of genomic alterations. Our series included 21 primary cancers and 19 cell lines, 14 of which have not been analyzed before. We found that normal pancreas expresses Fhit protein in the ductular region and that 42% of the cell lines and 62% of the primary cancers exhibit various alterations affecting FHIT gene expression.
MATERIALS AND METHODS
Normal Pancreas and Tumor Samples.
Samples of two normal pancreas were obtained from a pancreatic explant to be used for transplantation of insulae of Langerhans and from a splenopancreasectomy following abdominal trauma. The 21 primary pancreatic cancer samples were collected between 1984 and 1997 at the Pathology Department of the University of Verona (Verona, Italy; Table 1
). All common-type ductal cancers were graded according to Kløppel (23)
. Cases included 9 moderately and 10 poorly differentiated neoplasms. The remaining two cancers were a mucinous noncystic cancer (PC10) and an adenosquamous carcinoma (PC11). The nine cancers numbered consecutively from PC-3 to PC-11 have been described previously (24)
.
|
Immunostaining.
Paraffin-embedded sections of primary cancers and normal pancreas were immunostained after antigen retrieval by boiling twice for 5 min each in 0.1 M citrate buffer (pH 6) in a microwave oven (600 W). Incubation with anti-GST-Fhit antibody (1:5000) was overnight at 37°C. After three washes, the slides were incubated with biotinylated antirabbit IgG for 20 min and, subsequently, with streptavidin-peroxidase complex for 20 min. Antibody localization was effected by 10 min of incubation with 3,3'-diaminobenzidine. The slides were lightly counterstained with hematoxylin. Immunostaining for cell lines was essentially the same with 2 h incubation for anti-GST-Fhit antibody after fixation of cytospin preparations in cold methanol (-20°C).
Immunoblot Analysis.
Western blotting analysis was performed on two normal pancreas and all cell lines, whereas the contamination from nonneoplastic cells prevents such analysis on primary cancers. Freshly frozen pancreatic tissue fragments were minced in dry ice and lysed in radioimmunoprecipitation assay buffer [150 mM NaCl, 20 mM HEPES (pH 7.5), 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 1 mM EDTA, 10 µg/ml chymostatin-leupeptin-aprotinin-pepstatin, 1 mM phenylmethylsulfonyl fluoride, and 1 mM DTT]. Cultured cells were directly lysed (2 x 107 cells/ml). Proteins (50 µg/lane) were fractionated by SDS-PAGE in a 20% gel, electroblotted onto nitrocellulose membrane (2.5 mA/cm2 for 30 min) and reacted with anti-GST-Fhit antibody 1:5000 according to standard procedures, using goat antirabbit horseradish peroxidase-conjugated immunoglobulin for detection of the primary antibody. The ECL Western detection system (Amersham, Milan, Italy) was used, and the blot was exposed to X-ray film for 1–60 min.
RT-PCR and cDNA Sequencing.
Reverse transcription was performed using oligo(18-dT)s and nested-PCR amplification of FHIT mRNA using the primers listed in Table 2
. Total RNA was prepared from cell lines, normal pancreata and primary cancers using the Ultraspec RNA Extraction Kit (Biotec Laboratories, Houston, TX) according to the manufacturer’s instructions. Reverse transcription was performed in a 20-µl final volume using 200 units of Moloney murine leukemia virus reverse transcriptase (Life Technologies, Inc., Bethesda, MD) and 1–2 µg of total RNA, according to the manufacturer’s instructions. One µl of cDNA was used for the amplification with primers 5U2 and 3D2, from FHIT exons 1 and 10, respectively. One µl of the first PCR product at a 1:20 dilution was used for a second amplification with primers 5U1 and 3D1, located in exons 3 and 10 of FHIT. The PCR conditions were as described in Ohta et al. (6)
. The products corresponding to the normally and abnormally sized amplification products were excised from the agarose gel and the DNA purified using the Agarose Gel DNA Extraction Kit (Boehringer Mannheim, Mannheim, Germany). The DNA fragments were cloned in the TA vector (TA cloning kit; Invitrogen, San Diego, CA), and individual clones were sequenced with the primer 5U1 and/or 3D1.
|
. Briefly, a portion of the PCR mixture was denatured and run on polyacrylamide gels, which were dried on filter paper and exposed to X-ray film at -80°C for 1 h with an intensifying screen. The DNA samples showing mobility shifts of single-stranded DNA fragments were sequenced directly. To enrich for mutated sequences, the shifted bands were eluted from the gel, amplified, and then sequenced with the appropriate primers (Table 2)
as described previously (27)
. The wild-type bands of the same cases were also sequenced as internal controls.
DNA Analysis.
The status of the FHIT locus was studied by amplification of four polymorphic CA microsatellite loci, as described previously (24
, 28)
. The microsatellite loci were the D3S1312, centromeric to FHIT exon 1, and the three intragenic D3S1481, D3S1300, and D3S1234 loci (6
, 29)
. The study was performed using high molecular weight DNA from all cell lines and frozen samples of 12 primary cancers. The latter were those cancers which could be enriched to a neoplastic cellularity of >60% by cryostat dissection, as described previously (24)
. They included the nine cases from PC3 to PC11 (24)
, PC14, PC16, and PC17. In those primary cancers that could not be cryostat-enriched and in those cryostat-enriched cases showing allelic loss, the FHIT locus was further studied using DNA extracted from microdissected paraffin-embedded tissues (28)
. For this purpose, 10-µm paraffin-embedded, hematoxylin-stained sections were soaked for 5 min in 10 mM Tris-HCl (pH 8.0)-1 mM EDTA (TE buffer) and 1% glycerol. Fifty to 200 cells from neoplastic and nonneoplastic areas were scraped from the sections using a 27-gauge needle and collected in 20-µl DNA extraction mix (TE buffer, 1% Tween 20, and 200 µg/ml proteinase K) that was kept overnight at 37°C on a rotating wheel and subsequently boiled for 10 min; 4 µl from each served as template for PCR amplification.
PCR Amplification of FHIT Exons.
Exon-specific PCR amplification was accomplished using intronic primers and reaction conditions described in Druck et al. (30)
. The primers were UR4 and i15'R (exon 1), iex2F and iex2R (exon 2), iex3F and iex3R (exon 3), iex4F and iex4R (exon 4), iex5F and iex5R (exon 5), iex6F and iex6R (exon 6), iex7F and 016 (exon 7), iex8F and iex8R (exon 8), and iex9F and iex9R (exon 9).
RESULTS
Strategy.
Two normal pancreata and a panel of 40 pancreatic ductal cancers, including 21 primary cancers and 19 cell lines, were studied. The two normal pancreata and cancer cell lines were evaluated for Fhit protein expression by Western blot and immunohistochemistry using rabbit anti-GST-Fhit antibody (30)
, for FHIT mRNA status by RT-PCR, for allelic deletions at chromosome 3p14.2 by PCR amplification of four microsatellite markers, and for FHIT gene status by PCR amplification of exons 3–9. The same comprehensive analysis was performed on the primary cancers, with the following variations. Fhit protein was evaluated by immunohistochemistry alone, RT-PCR was performed in the only six cases for which RNA was available, and FHIT gene exon amplification was restricted to the nine cases for which it was possible to microdissect cancer cells or glands completely devoid of any contaminating nonneoplastic cells.
Normal Pancreas and Cancer-associated Pancreatitis.
Immunohistochemistry on normal pancreatic tissues using anti-GST-Fhit antiserum showed a cytoplasmic reactivity in the intralobular ductuli and centroacinar cells. In interlobular and larger size ducts and insulae of Langerhans as well as in acinar cells, Fhit expression was not detected (Fig. 1)
. However, the epithelium of large ducts and the insulae of Langerhans showed cytoplasmic Fhit expression in the large majority of chronic obstructive pancreatitis areas associated with cancer (Fig. 1C)
.
|
Cancer Cell Lines.
The results of the analysis on the cancer cell lines are summarized in Table 3
. Western blot analysis showed that Fhit protein was expressed at various levels in 11 cell lines and was undetectable in 8 (42%; Fig. 2
). Immunohistochemical and Western blot data matched for the 10 cell lines for which both analyses were performed.
|
|
. The PACA3 cell line lacking Fhit protein showed a shorter RT-PCR product that lacked exon 5, as revealed by sequence analysis. The normally sized RT-PCR products were not further analyzed, except in the case of ASPC1 and BJ-SW cells, both of which showed a similar abnormal pattern on SSCP analysis. Sequencing of cloned RT-PCR products of these two cell lines showed that their transcripts contained an insertion of the same 5'-GCTTAAG-3' sequence at the same nucleotide position within the noncoding exon 10 (439-CAGTGACACAGGCTTAAGATGTTTTTCAGATCC-465; 6
). In ASPC1, the abnormal transcript coexisted with a normal transcript. Both these cell lines expressed Fhit protein, and therefore, these insertions did not affect FHIT expression.
|
. The loss of exon 5 was confirmed by Southern blotting analysis (data not shown).
|
). Obviously, this comparative analysis could not be performed in all of the other cell lines. However, five cell lines exhibited homozygous loss of the D3S1300 marker in intron 5. In addition, the finding of only one allelic form in all four microsatellite markers in most cell lines suggests that this may be due to the loss of the second allele in a number of cases, considering the high degree of informativeness of these markers (see also Table 4
).
|
. Protein expression was not analyzed by Western blot in primary pancreatic cancers, due to the high admixture of contaminating nonneoplastic inflammatory and stromal cells. However, the coincidence of immunohistochemical and Western blot data in cell lines implied that immunohistochemistry was a reliable tool to investigate Fhit protein expression. The staining in primary tumors was considered as positive when the intensity of staining was well over the stromal background, which was used as the internal negative control, whereas the ductules served as internal positive controls. We scored as negative the cases with over 95% of cancer cells unstained. Thirteen of the 21 (62%) primary cancers showed the absence of Fhit protein and 8 cases (38%) showed Fhit positive neoplastic populations with variable intensity of staining (Fig. 1)
. In 4 of these 8 cases the large majority of cancer cells was stained, whereas a mixed population of anti-Fhit reactive and nonreactive neoplastic cells was observed in the remaining 4.
RNA was available for six primary cancers lacking Fhit and was analyzed by RT-PCR. Three cases showed normal sized FHIT RT-PCR products (PC-19, PC-23 and PC-24) and three cases showed shorter bands in addition to the normal one (PC-20, PC-21 and PC-22; Fig. 5
). Due to the absence of Fhit protein in the cancer cells of the samples showing aberrant mRNAs, we assumed that all of the altered transcripts were functionally equivalent, i.e., leading to the lack of Fhit protein expression. Therefore we characterized the single abnormal band found in case PC-22 and only the apparently more intense lower bands of cases PC-20 and PC-21. These bands were excised from the gel, cloned and sequenced. The sequencing revealed absence of exons 4 to 7, 5 to 7 and 5 to 6 in PC-20, PC-21 and PC-22, respectively, always accompanied by the lack of nucleotides 450–460. The normal sized product from all six cases was analyzed by SSCP and sequencing. This analysis showed that in five cases there was a mixed population of both full length and shorter transcripts lacking cDNA nucleotides 450–460 in the non coding exon 10 (439-CAGTGACACAGATGTTTTTCAGATCC-465), whereas the last case (PC-23) showed only the full length transcript. In addition, case PC-24 contained two polymorphisms, one at at codon 13 (TCT to TCC) and the second at codon 83 (CAG to CAA).
|
The search for the presence of FHIT exons was performed in the nine primary cancers in which it was possible to microdissect cancer cells or glands completely devoid of any contaminating nonneoplastic cell. These nine cases included four expressing and five lacking Fhit protein (see Table 4)
. The search was limited to exons 5 and 8, because the former was the commonly deleted exon in our cell lines, whereas the latter was never lost. Three of the five cases lacking Fhit protein showed homozygous deletion of exon 5 (Fig. 6)
, whereas both exons were present in the remaining six cases.
|
In this study, we demonstrated the following. (a) Fhit protein is constantly expressed in the intralobular ductules of the normal pancreas. Its expression was undetected in the larger ducts and insulae of Langerhans, where it may be induced by a chronic inflammatory status such as the cancer-associated obstructive pancreatitis; acinar cells never showed Fhit expression. (b) Fhit protein is expressed in a proportion of pancreatic cancers, both cell lines and primaries; those cancers lacking Fhit protein expression also show either the absence or the alteration of FHIT mRNA, which is often associated with FHIT gene anomalies.
In the normal pancreas, only ductular cells expressed large amounts of Fhit protein in the cytoplasm. The absence of the protein in interlobular and larger ducts implies the existence of a specific metabolic status of the ductal epithelium in the more distal district that requires Fhit protein expression. It is notable that only 60% of the cancer cell lines and 40% of primary cancers expressed various amounts of Fhit protein. This suggests that, if most of these cancers derived from the Fhit-positive distal ductal epithelium, they have lost Fhit expression during cancer development or progression, consistent with the genomic lesions in the FHIT gene in Fhit-negative cancers.
All of the 11 cancer cell lines expressing Fhit protein did not show alterations in the mRNA open reading frame. The only alterations were detected in the noncoding exon 10 and did not affect Fhit expression. They consisted in a novel 7-bp insertion (5'-GCTTAAG-3') at position 449 of the cDNA in two cell lines (ASPC1 and BJ-SW), and in the deletion of 11 bp at the beginning of exon 10 and including cDNA nucleotides 450–460 in all 11 cell lines. This latter represents an alternatively spliced form (FHITß) that has been described previously (11 , 21 , 31) . On the other hand, our analysis showed that in eight cancer cell lines the lack of Fhit protein expression was always associated with the absence or the abnormality of FHIT mRNA. A genomic lesion was demonstrated in five of these eight cell lines and consisted in a homozygous loss at chromosome 3p14.2 encompassing the D3S1300 locus and a variable number of FHIT exons, always including exon 5. The occurrence of these lesions as a cell culture artifact could be ruled out by our analysis of primary cancers.
Fhit protein was expressed in 40% of primary cancers. Our finding that normal pancreatic large ducts express Fhit protein in the presence of chronic pancreatitis suggests that the expression of Fhit protein in pancreatic cancer cells might be caused by coexistent inflammatory stimuli. However, the large majority of Fhit protein-negative primary pancreatic cancers were associated with chronic pancreatitis changes as well. This makes it more likely that the lack of Fhit protein might represent a defect in genetic information rather than a metabolic condition leading to Fhit underexpression. The possibility to record the lack of FHIT mRNA in primary pancreatic cancers is prevented by the common contamination from nonneoplastic tissue. By RT-PCR analysis, three of six Fhit protein-negative cases presented one or two shorter transcripts in addition to the normally sized transcript. However, the occurrence of more than one shorter transcript in some cases together with the fact that these were characterized by the lack of a variable array of entire exons suggests that aberrant splicing events may be the cause of this phenomenon (32 , 33) . Therefore, the question whether the lack of Fhit protein was due to the lack of genetic information could only be addressed by the analysis of cancer cell DNA. This is particularly difficult for pancreatic cancer due to the paucity of cancer cells in this type of malignancy. We overcome such difficulty using cryostat enrichment of samples to a neoplastic cellularity of at least 60%. The analysis of this material showed that the allelic loss of microsatellite DNA markers internal to the FHIT gene occurred in the large majority (10 of 13; 77%) of primary cancers lacking Fhit protein expression, but in only 1 of the 8 (12%) cancers expressing Fhit protein. Considering the low genomic resolution of this analysis, there was a good correlation between loss of specific DNA markers internal to the FHIT locus and lack of Fhit protein expression.
A closer genomic analysis could be performed in five primary cancers lacking Fhit protein expression for which it was possible to obtain pure cancer DNA by microdissection of paraffin-embedded samples. The finding of FHIT exon 5 homozygous deletions in three of the five cases demonstrates that a genetic lesion is responsible for the absence of Fhit protein in these cancers. Our data also suggest that homozygous loss of FHIT exons, either in cell lines or primary cancers, is almost always associated with the homozygous loss of the D3S1300 marker internal to the gene. It has been reported that in some primary tumors and cancer cell lines homozygous deletions of intronic regions may occur and still leave intact FHIT exons (6 , 34) . On the other hand, one of our primary cancers (PC-6) illustrates that homozygous loss of FHIT exons may occur without a corresponding homozygous loss of intronic regions, as detected by microsatellite marker analysis.
Interestingly, the presence of exon 5 in two Fhit protein-negative microdissected cancers (PC-17 and PC-18) suggests integrity of the FHIT gene in these cancers, as observed for three cell lines (SK-PC1, PANC1, and MIAPACA2). These cell lines did not express detectable amounts of Fhit protein and transcripts, despite the presence of FHIT exons 3–9, suggesting that mechanisms other than homozygous exon loss may be involved in lack of Fhit expression in these cases. It cannot be excluded, however, that independent nonoverlapping deletions of the two FHIT alleles, such as reported for KatoIII and other cell lines (16 , 35) could have inactivated both FHIT alleles in these tumors.
In conclusion, our study shows that FHIT gene alterations, represented by exon loss and/or intron alterations that may affect mRNA stability or expression, occur in a significant proportion of pancreatic primary ductal adenocarcinomas. Whether these represent a causative or secondary event in pancreatic cancer pathogenesis remains an open issue, which is beyond the scope of this study.
ACKNOWLEDGMENTS
We thank Dr. A. Achille for performing some of the sequencing reactions.
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 This study was supported by grants from the Associazione Italiana Ricerca Cancro (Milan, Italy; to A. S.); Consorzio Studi Universitari and Banca Popolare di Verona (Verona, Italy); BIOMED 2 CE-Contract BMH4-CT98-3805; Ministero Università e Ricerca Scientifica e Tecnologica (Rome, Italy); and United States Public Health Service Grants CA21124 and CA56336 from the National Cancer Institute. 
2 To whom requests for reprints should be addressed, at Dipartimento di Patologia-Anatomia Patologica, Università di Verona, Strada Le Grazie, 8, I-37134 Verona, Italy. Phone: (39)(45) 8098 617; Fax: (39)(45) 8098 136; E-mail: scarpa{at}anpat.univr.it
3 The abbreviations used are: RT-PCR, reverse transcription-PCR; GST, glutathione S-transferase; LOH, loss of heterozygosity; SSCP, single-strand conformation polymorphism.
Received 10/ 2/98. Accepted 1/13/99.
REFERENCES
This article has been cited by other articles:
|
|
 |
|
  M. Takahashi, T. Kitahashi, R. Ishigamori, M. Mutoh, M. Komiya, H. Sato, Y. Kamanaka, M. Naka, T. Maruyama, T. Sugimura, et al. Increased expression of inducible nitric oxide synthase (iNOS) in N-nitrosobis(2-oxopropyl)amine-induced hamster pancreatic carcinogenesis and prevention of cancer development by ONO-1714, an iNOS inhibitor Carcinogenesis, August 1, 2008; 29(8): 1608 - 1613. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Takeuchi, M. Takahashi, K. Sakano, M. Mutoh, N. Niho, M. Yamamoto, H. Sato, T. Sugimura, and K. Wakabayashi Suppression of N-nitrosobis(2-oxopropyl)amine-induced pancreatic carcinogenesis in hamsters by pioglitazone, a ligand of peroxisome proliferator-activated receptor {gamma} Carcinogenesis, August 1, 2007; 28(8): 1692 - 1696. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Trapasso, S. Yendamuri, K. R. Dumon, R. Iuliano, R. Cesari, B. Feig, R. Seto, L. Infante, H. Ishii, A. Vecchione, et al. Restoration of receptor-type protein tyrosine phosphatase {eta} function inhibits human pancreatic carcinoma cell growth in vitro and in vivo Carcinogenesis, November 1, 2004; 25(11): 2107 - 2114. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. I. Wistuba, R. Ashfaq, A. Maitra, H. Alvarez, E. Riquelme, and A. F. Gazdar Fragile Histidine Triad Gene Abnormalities in the Pathogenesis of Gallbladder Carcinoma Am. J. Pathol., June 1, 2002; 160(6): 2073 - 2079. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Bodmer, M. Eleveld, E. Kater-Baats, I. Janssen, B. Janssen, M. Weterman, E. Schoenmakers, M. Nickerson, M. Linehan, B. Zbar, et al. Disruption of a novel MFS transporter gene, DIRC2, by a familial renal cell carcinoma-associated t(2;3)(q35;q21) Hum. Mol. Genet., March 1, 2002; 11(6): 641 - 649. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 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]
|
|
|
|
|
|
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]
|
|
|
|
 |
|
 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]
|
|
|
|
 |
|
 K. M. Anderson and J. E. Harris Selected Features of Nonendocrine Pancreatic Cancer Experimental Biology and Medicine, June 1, 2001; 226(6): 521 - 537. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. R. Dumon, H. Ishii, A. Vecchione, F. Trapasso, G. Baldassarre, F. Chakrani, T. Druck, E. F. Rosato, N. N. Williams, R. Baffa, et al. Fragile Histidine Triad Expression Delays Tumor Development and Induces Apoptosis in Human Pancreatic Cancer Cancer Res., June 1, 2001; 61(12): 4827 - 4836. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. S. Moore, G. Zamboni, A. Brighenti, D. Lissandrini, D. Antonello, P. Capelli, G. Rigaud, M. Falconi, and A. Scarpa Molecular Characterization of Pancreatic Serous Microcystic Adenomas : Evidence for a Tumor Suppressor Gene on Chromosome 10q Am. J. Pathol., January 1, 2001; 158(1): 317 - 321. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Ouyang, L.-j. Mou, C. Luk, N. Liu, J. Karaskova, J. Squire, and M.-S. Tsao Immortal Human Pancreatic Duct Epithelial Cell Lines with Near Normal Genotype and Phenotype Am. J. Pathol., November 1, 2000; 157(5): 1623 - 1631. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Mori, K. Mimori, T. Shiraishi, H. Alder, H. Inoue, Y. Tanaka, K. Sugimachi, K. Huebner, and C. M. Croce Altered Expression of Fhit in Carcinoma and Precarcinomatous Lesions of the Esophagus Cancer Res., March 1, 2000; 60(5): 1177 - 1182. [Abstract] [Full Text]
|
|
|
|
|
|
R. Baffa, L. G. Gomella, A. Vecchione, P. Bassi, K. Mimori, J. Sedor, C. M. Calviello, M. Gardiman, C. Minimo, S. E. Strup, et al. Loss of FHIT Expression in Transitional Cell Carcinoma of the Urinary Bladder Am. J. Pathol., February 1, 2000; 156(2): 419 - 424. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. P. Hao, J. E. Willis, T. G. Pretlow, J. S. Rao, G. T. MacLennan, I. C. Talbot, and T. P. Pretlow Loss of Fragile Histidine Triad Expression in Colorectal Carcinomas and Premalignant Lesions Cancer Res., January 1, 2000; 60(1): 18 - 21. [Abstract] [Full Text]
|
|
|
|
 |
|
 K. Mimori, T. Druck, H. Inoue, H. Alder, L. Berk, M. Mori, K. Huebner, and C. M. Croce Cancer-specific chromosome alterations in the constitutive fragile region FRA3B PNAS, June 22, 1999; 96(13): 7456 - 7461. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Y. Y. Fong, V. Fidanza, N. Zanesi, L. F. Lock, L. D. Siracusa, R. Mancini, Z. Siprashvili, M. Ottey, S. E. Martin, T. Druck, et al. Muir-Torre-like syndrome in Fhit-deficient mice PNAS, April 25, 2000; 97(9): 4742 - 4747. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 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 |
x174 DNA.
