Squamous Cell Carcinoma and Adenocarcinoma Display Similar Expression Patterns |
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
Abstract |
|---|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
Highly glycosylated apomucins are important to maintain the viscoelastic properties of the mucus. Changes in their expression are frequently associated with inflammatory and neoplastic conditions. We analyzed the expression of apomucins in normal respiratory tract (n = 8) and compared it with distal, peritumoral, and tumoral epithelia from patients with squamous cell carcinoma (n = 20), adenocarcinoma (n = 13), and small cell carcinoma (n = 12). Squamous metaplasia (n = 16) was also analyzed. MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC8 apomucins were detected by immunohistochemistry, and mucin transcripts by in situ hybridization and reverse transcriptase polymerase chain reaction. Bronchial epithelium from normal individuals and distal epithelium from cancer patients showed a similar expression pattern: MUC1, MUC4, and MUC8 were always present, MUC2 and MUC5AC showed more variability, and MUC6 was focally detected. MUC5AC was downregulated in peritumoral epithelium and in squamous metaplasia, and MUC6 was upregulated in peritumoral epithelium. A reduced expression of MUC4, MUC5AC, and MUC8 was observed in non-small cell carcinomas, regardless of their histologic subtype. In small cell tumors, only MUC1 was consistently expressed. Conclusions: (1) peritumoral epithelium and squamous metaplasia show an abnormal pattern of mucin expression; (2) squamous cell carcinomas and adenocarcinomas display a similar pattern of mucin gene expression, supporting the concept of a common cellular origin.
| |
Introduction |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
Mucus covering the respiratory tract plays an important role in the protection from external aggressions such as solid particles, pathogens, and chemical agents. The viscoelastic properties of mucus are mainly determined by the presence of mucins, which are high molecular weight proteins that are extensively O-glycosylated. Nine genes coding for human mucins have been cloned (MUC1 through MUC4, MUC5AC, MUC5B, and MUC6 through MUC8) (1). Among them, MUC4, MUC5AC, MUC5B, and MUC8 were cloned from tracheobronchial complementary DNAs (cDNAs). MUC1, MUC3, and MUC4 are transmembrane molecules (2). MUC7 gene encodes the human low-molecular weight salivary mucin (5). A partial cDNA sharing some characteristics with the mucin genes has been named MUC8 (6) and is detected in epithelial cells from pluri- and monostratified epithelia (7). MUC2, MUC5B, MUC5AC, and MUC6 genes form a cluster located in the 11p15 chromosomal region; they code for secreted gel-forming mucins containing cysteine-rich domains homologous to the D domain of von Willebrand factor (8). Transmembrane-type mucins are often detected in cells that do not contain mucus droplets (2, 4). By contrast, gel-forming mucins are generally detected in goblet or goblet-like cells. For example, in the intestine, MUC2 is present in goblet cells but not in absorptive cells (9). In the tracheobronchial epithelium, mucins are synthesized by the goblet cells in the surface epithelium and the mucous cells in the submucosal glands.
The expression pattern of mucin genes has been reported to be altered in several pathologies such as cancer
(10). The mechanisms leading to the ectopic expression
or downregulation of mucin genes remain at this moment
unclear, partly because there is little information regarding the regulatory sequences of mucin genes. Recently,
some evidence regarding the molecular pathways involved in the control of mucin gene expression has been published. Pseudomonas aeruginosa, a major pathogen in patients with cystic fibrosis (CF), activates the transcription
of MUC2 through the Src-Ras-MEK1/2-ERK1/2-pp90rsk-NF-
B pathway (14). The exoproducts of this pathogen
also activate the transcription of MUC5AC, and the elements responsible for the activation of the reporter have
been identified within 4 kb of the transcriptional start site
(15). In addition to changes in apomucin expression, glycosylation has also been shown to be altered in several respiratory diseases. Variations in both length and charge of
O-linked mucin carbohydrate chains may alter the physicochemical properties of mucus. These alterations can be originated by changes in the expression and/or abnormal
subcellular localization of glycosyltransferases and glycosidases, as it has been described in several cancer cells (16).
In respiratory diseases such as CF, chronic bronchitis, and
asthma (17), changes in the viscoelasticity of the mucus
play an important role in the pathogenesis of the disease.
In this study, we have analyzed the expression of MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC8 in different types of lung tumors as well as in their peritumoral and normal distal epithelium. Our results show that non-small cell lung cancer (NSCLC) is associated with specific changes in apomucin expression, and that squamous cell carcinoma and adenocarcinoma, but not small cell lung cancer (SCLC), show a remarkably similar pattern of apomucin expression.
| |
Materials and Methods |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
Patients and Tissue Samples
Patients were recruited at Hospital del Mar between May 1996 and April 1998. Sixty-four patients with lung cancer who underwent a fiberoptic bronchoscopy for diagnostic purposes were included in the study. Normal tissue samples were obtained from all of them; whenever possible, tumor samples were also obtained for analysis (Table 1). Control samples (n = 8) were obtained from nonsmoking individuals with normal lung function undergoing bronchoscopy for diagnostic purposes in whom no detectable bronchial or lung pathology was found. Squamous metaplasia samples (n = 16) were obtained from patients with chronic obstructive pulmonary disease (COPD) (n = 4) or with lung cancer (n = 12). Bronchoscopy samples were embedded in paraffin and used for diagnostic and immunohistochemical assays. Histologic diagnoses were made by consensus of two independent pathologists with extensive experience in lung cancer classification (C.B. and J.L.) following standard criteria. For RNA analysis, samples were collected from cases undergoing elective surgery. Tissue was frozen in liquid nitrogen or immediately fixed in 4% paraformaldehyde. Informed consent was obtained from patients and the institution's Ethics Committee approved the study.
|
Antibodies
The characteristics of the antiapomucin antibodies used in this study are described in Table 2. BC-3 (18) and LDQ10 (9) monoclonal antibodies (mAb) were used as ascites fluid diluted at 1:250. As negative control, the B12 mAb recognizing synthetic dextran (R. Castro, Barcelona, Spain) was used. Rabbit polyclonal antiserum raised against rat gastric mucin (RGM) and recognizing human MUC5AC (19) was used at 1:200 (A. Einhard, Rotterdam, The Netherlands). Polyclonal anti-MUC4 (20), anti-MUC6.1 (21), and anti-MUC8 antibodies were purified by affinity chromatography on the synthetic peptides coupled to AH-sepharose 4B (Pharmacia, Uppsala, Sweden). For MUC4 and MUC6, peptides corresponding to the tandem repeat (TR) sequences were used, whereas for MUC8 a peptide from the COOH-terminus was selected (GTPGSGLLPAHIVPLSKSEER) (6). The specificity of all of the antibodies has been previously reported (Table 2) (22), except for MUC8. The recognition of the mucin precursor has been demonstrated for the anti-MUC2 (9), anti-MUC4, and anti-MUC6 antibodies that recognize the TR sequences of these apomucins. Anti-MUC5AC polyclonal antibodies, raised against the unique sequence, recognize the precursor and mature forms of this apomucin (22). Specificity of the anti-MUC8 affinity-purified antibodies was determined by enzyme-linked immunosorbent assay and by peptide inhibition assays by immunohistochemistry on bronchial sections as previously described (13). Anti-MUC4 antibodies were used at 1:50 dilution and anti-MUC6.1 and anti-MUC8 antibodies were used at 1:10. Pre-immune rabbit serum was used as negative control at 1:500 dilution. Periodic acid-Schiff (PAS) staining was performed following standard procedures, including diastase digestion of tissue sections.
|
Immunohistochemistry
The indirect immunoperoxidase technique was performed on 5-µm sections of paraffin-embedded tissues. Samples were dewaxed, rehydrated, and fixed in cold acetone for 10 min. Endogenous peroxidase was blocked by immersion in 4% hydrogen peroxide for 10 min. Nonspecific binding sites were blocked with 5% skim milk in phosphate-buffered saline (PBS). Primary antibodies were diluted in 1% PBS-bovine serum albumin and applied for 1.5 h. After washing the slides with PBS, peroxidase-labeled secondary antibodies (DAKO, Glostrup, Denmark) were incubated for 1 h. Peroxidase reaction was developed using 0.5% diaminobenzidine with 0.1% H2O2 in PBS. The slides were counterstained with hematoxylin, dehydrated, and mounted with DPX (BDH, Poole, UK).
In Situ Hybridization
Tissue samples were immediately fixed in 4% paraformaldehyde, embedded in paraffin, and stored at 4°C. Sense and antisense synthetic oligonucleotides (48 bp) corresponding to the TR sequence of different mucin genes were labeled with digoxigenin following the manufacturer's instructions (Boehringer Mannheim, Mannheim, Germany). The sequences corresponding to the antisense oligonucleotides were the following: MUC2: 5' GGT CTG TGT GCC GGT GGG TGT TGG GGT TGG GGT CAC CGT GGT GGT GGT 3'; MUC4: 5' GTC GGT GAC AGG AAG AGG GGT GGC GTG ACC TGT GGA TGC TGA GGA AGT 3'; MUC5AC: 5' AGG GGC AGA AGT TGT GCT CGT TGT GGG AGC AGA GGT TGT GCT GGT TGT 3'; MUC6: 5' CAT CTG TGC GTG GGT AGG GGT GAT GAC TGT GTG AGT ACT TGG AGT CAC 3'. The in situ hybridization technique was performed as described elsewhere (20). The following tissues were used as controls: normal colon (MUC2 and MUC4) and normal stomach (MUC5AC and MUC6) (13, 20).
Reverse Transcriptase Polymerase Chain Reaction
Total RNA was isolated using the Chomczynski method. cDNA was synthesized using 5 µg of DNAse I-treated RNA with 200 U Moloney-Murine Leukemia Virus reverse transcriptase (RT). The primers and conditions for MUC5AC and MUC6 amplification were described elsewhere (20). Primers for MUC2 were: sense, CTT CGA CGG ACT CTA CTA CAG C and antisense, CTT TGG TGT TGT TGC CAA AC. MUC4 transcripts were detected using ASPG-1 primers: sense, CTT ACT CTG GCC AAC TCT GTA GTG; antisense, GAG AAG TTG GGC TTG ACT GTC; and MUC8 was detected using published primers (23). The conditions for amplification were, for MUC2, 1 min at 94°C, 30 s at 58°C, 30 s at 72°C (35 cycles); for MUC4, 30 s at 94°C, 1 min at 55°C, and 1 min at 72°C (30 cycles); and for MUC8, 30 s at 94°C, 45 s at 60°C, 1 min at 72°C (40 cycles). K-ras primers were used to control for messenger RNA (mRNA) quality and integrity. The size of the polymerase chain reaction (PCR) products was 387 bp for MUC2, 468 bp for MUC4, 240 bp for MUC8, and 265 bp for K-ras. Conditions for MUC5AC and MUC6 were previously described (20). mRNAs from normal colon and stomach were used as positive control for MUC2 and MUC4, and mRNA from normal stomach was used as control for MUC5AC and MUC6, respectively.
Statistical Analysis
Values are expressed as mean ± standard deviation (SD) or as
positive cases/total cases. For the statistical analysis, the Fisher's
exact test was used. Statistical significance was defined as P 
0.05.
| |
Results |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
Apomucin Expression in Normal and Peritumoral Bronchial Epithelium
Bronchoscopy samples obtained from an unaffected lobar bronchus ipsilateral to the tumor were considered as distal samples. Bronchial epithelium was considered peritumoral when it was located in the same tissue sample as the tumor and no metaplastic changes were identified. Because there were no differences in the apomucin expression in distal or peritumoral epithelium among patients with various types of lung cancer, only the pooled results are presented here. MUC1, MUC4, and MUC8 were detected in all samples of nontumoral bronchial epithelia (Table 3). These apomucins were found in both mucus-secreting and ciliated cells. MUC2, expressed in the goblet cells and focally in ciliated cells, was variably detected in both distal and peritumoral epithelia (11/18 versus 28/40, respectively). The expression of MUC5AC in the vacuoles of mucus-secreting cells was lower in the peritumoral than in the distal epithelium (23/ 35 versus 15/16) and this difference was statistically significant (P = 0.04). MUC6 was detected focally in distal and peritumoral epithelia in approximately 20% of the samples. Examples of these results are shown in Figure 1.
|
|
These results are similar to those obtained using control samples (n = 8) from nonsmokers with normal lung function without bronchial or lung pathology (Tables 1 and 3). In these samples, the pattern of stained cells was not different from that found in distal and peritumoral epithelia. MUC1, MUC4, and MUC8 were highly expressed in all the samples analyzed. MUC2 was variably detected in the samples analyzed (5 to 70% of positive cells), and MUC5AC was found in the goblet cells (30 to 100%) of all the control samples. MUC6 was undetectable.
Therefore, normal and distal tissue display essentially identical apomucin expression patterns, whereas peritumoral epithelium is characterized by a downregulation of MUC5AC and an upregulation of MUC6 apomucins.
Apomucin Expression in Squamous Metaplasia
Squamous metaplasia was present in some biopsies (n = 16) from patients with neoplasia (n = 12) or smokers with COPD (n = 4) (Table 1). Apomucin expression was similar in squamous metaplasia and in peritumoral epithelium (Table 3). MUC1, MUC4, and MUC8 were detected in all the analyzed cases in a high proportion of cells (30 to 100%), MUC2 and MUC5AC were found in a broad range of cells (10 to 100%), and MUC6 was only detected in one case (20% of positive cells). No differences were found regarding the pathologic status of the patient (COPD or lung cancer; data not shown). When this apomucin expression pattern was compared with that found in normal samples, only the expression of MUC5AC was significantly decreased (P = 0.04). Representative examples of these results are shown in Figure 2.
|
Apomucin Expression in Squamous Cell Carcinoma
The tumors from 20 patients with squamous cell carcinoma were studied. All of them were unreactive with PAS-diastase staining. MUC1, MUC4, and MUC8 apomucins
were detected in 
75% of tumors (40 to 100, 5 to 100, and
50 to 100% of positive cells, respectively). The proportion
of samples, and cells within a given sample, expressing MUC2 showed greater variation. MUC5AC and MUC6
were detected in < 30% of tumors (Table 3). When the
pattern of apomucin expression in squamous cell carcinoma was compared with that of peritumoral epithelium,
only the expression of MUC4 was significantly lower (P = 0.03). Representative results are shown in Figures 3 and 4.
|
|
Apomucin Expression in Lung Adenocarcinoma
The tumors from 13 patients with lung adenocarcinoma were analyzed. Ten of 11 tumors examined were reactive with PAS-diastase staining, with a proportion of positive cells ranging from 5 to 60%. MUC1 was detected in all samples, and MUC4 and MUC8 were detected in approximately two-thirds of them, with a range of 50 to 100, 5 to 100, and 20 to 100% positive cells, respectively. Expression of MUC4 and MUC8 was independent of each other. MUC2 and MUC5AC were detected in one-third or less of the cases (10 to 80 and 5 to 70% of positive cells), and MUC6 was absent from these tumors. When the pattern of apomucin expression in adenocarcinoma was compared with that of peritumoral epithelium, a reduced expression of MUC8 (P = 0.03) and MUC5AC (P = 0.01) was observed. Representative results are shown in Figure 5.
|
Apomucin Expression in Small Cell Lung Carcinoma
MUC1 was the only apomucin that was commonly detected in SCLC (10 of 12 cases). MUC4 was detected in 3 of 12 cases, whereas the other apomucins were generally undetectable (Table 3). MUC6 was not detected in any of the tumors. The expression of MUC2, MUC4, MUC5AC, and MUC8 in small cell carcinomas was significantly different from that found in the peritumoral bronchial epithelium (P < 0.05).
Detection of Mucin Transcripts by In Situ Hybridization and RT-PCR
To confirm the immunohistochemical findings described previously using immunohistochemistry, RNA analysis using in situ hybridization and RT-PCR was carried out on selected paired samples of normal and tumor tissues. By in situ hybridization, MUC2 mRNA was not detected in samples from two cases (one adenocarcinoma and one squamous cell carcinoma) analyzed. MUC4 transcripts were found in 6 of 7 samples of normal distal epithelium, in 3 of 3 adenocarcinomas, and in 2 of 4 squamous cell carcinomas. MUC5AC was expressed in 3 of 5 samples of normal distal mucosa and it was undetectable in five tumors (four adenocarcinomas and one squamous cell carcinoma) (Figure 6). MUC6 mRNA was not detected in the distal epithelium (n = 3) nor in tumor samples (one adenocarcinoma and two squamous cell carcinomas) examined.
|
Similar results were obtained by RT-PCR (Table 4). In normal distal bronchus from lung cancer patients, MUC4, MUC5AC, and MUC8 mRNAs were detected, whereas MUC2 and MUC6 transcripts were absent. In tumor samples, MUC8 and MUC4 mRNAs were commonly detected, whereas MUC5AC and MUC6 were found only in one adenocarcinoma sample, and MUC2 was not detected. Representative results are shown in Figure 7.
|
|
| |
Discussion |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
In normal tracheobronchial epithelium, MUC1, MUC2, MUC4, MUC5B, MUC5AC, MUC7, and MUC8 apomucins show a cell-specific distribution (6, 24). Here, we have analyzed the pattern of mucin gene expression in bronchial epithelium from normal subjects and compared it with that of distal and peritumoral bronchial epithelia from lung cancer patients. The pattern of mucin gene expression detected in control samples was indistinguishable from that found in the distal epithelium of patients with cancer. By contrast, peritumoral epithelium and squamous metaplasia showed distinct changes: MUC5AC is downregulated in both types of lesions and MUC6 is upregulated in peritumoral epithelium. Because squamous metaplasia is a premalignant lesion that can be induced by lung carcinogenic agents (27), downregulation of MUC5AC may constitute a marker of risk of lung cancer development. In cancer samples, MUC4 was downregulated in the squamous cell carcinomas, whereas MUC5AC and MUC8 were downregulated in adenocarcinomas. In SCLC samples, MUC1 was the only apomucin commonly detected. MUC1, considered a marker for the epithelial lineage, is ubiquitously expressed in epithelial cells (28). Regarding apomucin expression in the two main types of NSCLC, the only difference found was that MUC8 expression occurred less frequently in adenocarcinomas than in squamous cell carcinomas.
It is thought that bronchogenic tumors arise from a pluripotential stem cell located in the basal bronchial epithelium. In SCLC, cells undergo neuroendocrine differentiation (29), whereas in squamous cell carcinoma and adenocarcinoma they acquire surface or glandular epithelial features, respectively (30). This common origin would explain why these tumors are closely related clinically and epidemiologically. The fact that SCLC displays epithelial and neuroendocrine markers may support the hypothesis of an early common precursor cell with NSCLC. Several genetic alterations have been reported to be associated with lung carcinogenesis. Recently, the loss of expression of Fhit has been reported to be the most frequent genetic change in lung cancer, especially in squamous tumors, and its loss can also be detected in preneoplastic lesions (31). By contrast, mutations in the K-ras gene are restricted specifically to adenocarcinomas and arise late in the pathogenesis of lung cancer (32).
Few specific markers for each type of NSCLC have
been identified. Squamous cell carcinomas can be distinguished from adenocarcinomas because they commonly
express SCCA1 and SCCA2 antigens, encoded by genes
that belong to the serpin superfamily (33). A remarkable finding of our studies was that squamous cell carcinomas
and adenocarcinomas displayed very similar patterns of
apomucin expression. By contrast, PAS-diastase staining
discriminated between adenocarcinomas (positive) and
squamous cell carcinomas (negative), supporting the idea
that apomucins synthesized in the adenocarcinoma cells
are highly glycosylated, whereas the same apomucins detected in squamous cell carcinoma cells are underglycosylated. However, more biochemical work is necessary to establish this notion. Several antibodies used in our study
were raised against the TR of the apomucins and do not react with glycosylated apomucins (9, 22). Because the
rate of polypeptide glycosylation and the proportion of
apomucin that is underglycosylated may vary among the
two subtypes of NSCLC and because mRNA expression
was not analyzed in a quantitative fashion, it is possible
that there are differences in the levels of expression of mucin genes that would not be apparent in these studies. For
example, in NSCLC, increased levels of MUC1, MUC3, MUC4, MUC5B, and MUC5AC transcripts have been detected (34), and recently higher expression levels of
MUC5AC have been reported in adenocarcinomas than in
squamous cell carcinomas (37, 38). As in this study, we
have previously reported, in pancreas cancer, the detection of MUC2 apomucin in the absence of detectable levels of mRNA using in situ hybridization and RT-PCR (12),
suggesting low levels of transcripts and intracellular accumulation of underglycosylated apomucins, as it has been
described for MUC5AC in benzyl-N-acetyl-
-D-galactosamidine-treated HT-29 MTX cells (39). The relationship
between the levels of mRNA and apomucin merits further
studies. Alterations in the glycosylation pathway in epithelial cells have been described in respiratory diseases. In
cells from patients with CF, a reduction in the sialylation rate of glycoproteins, as well as an altered ligand traffic as a consequence of a defective acidification of the trans-Golgi, trans-Golgi network, prelysosomes, and endosomes
(40), has been described. In these cells, it has also been
suggested that dysfunctional intracellular CF transmembrane conductance regulator may alter endosomal pH,
leading to changes in glycoprotein biosynthesis that will be
reflected in their carbohydrate composition (41). In O-glycosylation, the sequential addition of monosaccharides is
catalyzed by specific glycosyltransferases resident in the Golgi cisternae. The first steps in this process are the synthesis of N · acetylgalactosanine-O-Ser/Thr (Tn), galactose-N-acetylgalactosanine-O-Ser/Thr (T), sialyl-Tn, and
sialyl-T core oligosaccharides that have been identified as
markers for several tumor types (42). In lung tumors, no
differences have been detected in the expression of T and
sialyl-T antigens in the two main types of NSCLC (43), whereas sialyl-Tn antigen is more commonly detected in
adenocarcinomas than in squamous cell carcinomas (44).
Terminal oligosaccharide chains, such as Lewis x, sialyl
Lewis x, and Lewis y, are also more frequently detected in
adenocarcinomas than in squamous cell carcinomas (45).
These data may suggest that each of the two main types of
NSCLC displays a specific set of fucosyl- and sialyltransferases. The characteristic pattern of glycosyltransferase
gene expression in normal tissues may be altered during
neoplastic transformation, as it has been described for fucosyltransferases and sialyltransferases in gastric and colon
epithelia (20, 46). Recently, Mandel and coworkers (47)
reported differences in the repertoires of three GalNAc-transferases in stratified epithelia. These GalNAc-transferases are different in different cell types as well as
changes in relation to cellular differentiation and in cancer. Further studies on the alterations of Golgi complex
structure (48), as well as in changes in the compartmentalization of the glycosyltransferases in the two main types of
NSCLC, would be helpful to better understand the molecular basis of mucin biosynthesis in these tumors.
| |
Footnotes |
|---|
Address correspondence to: Carme de Bolós, Unitat de Biologia Cel·lular i Molecular - IMIM, Dr. Aiguader, 80, 08003 Barcelona, Spain. E-mail: cbolos{at}imim.es
(Received in original form July 11, 2000 and in revised form October 5, 2000).
Acknowledgments: The authors thank M. Pavesi for statistical analyses, A. Einhard and R. Castro for the RGM and B12 antibodies, K. L. Carraway for the MUC4 primers, D. Swallow, J. Minguella, J. Broquetas, and M. Orozco-Levi for valuable contributions, M. Escorihuela, A. Roig, and N. Soler for excellent technical assistance, and L. Español for help in the preparation of the manuscript. This study was supported by grants 96/1703 and 00/0610 from the Fondo de Investigación Sanitaria, grant SAF97-0085 from the Comisión Interministerial de Ciencia y Tecnología, grant 1997SGR-00078 from CIRIT (Generalitat de Catalunya), grant BMH4-CT98-3222 from Biomed, SEPAR, and FUCAP. A.L.-F is a predoctoral fellow from CIRIT.
Abbreviations cDNA, complementary DNA; CF, cystic fibrosis; mRNA, messenger RNA; NSCLC, non-small cell lung cancer; PAS, periodic acid-Schiff; PBS, phosphate-buffered saline; RT-PCR, reverse transcriptase polymerase chain reaction; SCLC, small cell lung cancer; TR, tandem repeat.
| |
References |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
1. Gendler, S. J., and A. P. Spicer. 1995. Epithelial mucin genes. Annu. Rev. Physiol. 57: 607-634 [Medline].
2.
Gendler, S. J.,
J. Taylor-Papadimitriou,
T. Duhig,
J. Rothbard, and
J. A. Burchell.
1988.
A highly immunogenic region of a human polymorphic epithelial mucin expressed by carcinomas is made up of tandem repeats.
J.
Biol. Chem.
263:
12820-12823
3. Williams, S. J., D. J. Munster, R. J. Quin, D. C. Gotley, and M. A. McGuckin. 1999. The MUC3 gene encodes a transmembrane mucin and is alternatively spliced. Biochem. Biophys. Res. Commun. 261: 83-89 [Medline].
4. Moniaux, N., S. Nollet, N. Porchet, P. Degand, A. Laine, and J. P. Aubert. 1999. Complete sequence of the human mucin MUC4: a putative cell membrane-associated mucin. Biochem. J. 338: 325-333 .
5.
Bobek, L. A.,
H. Tsai,
A. R. Biesbrock, and
M. J. Levine.
1993.
Molecular
cloning, sequence, and specificity expression of the gene encoding the low
molecular weight human salivary mucin (MUC7).
J. Biol. Chem.
268:
20563-20569
6. Shankar, V., M. S. Gilmore, R. C. Elkins, and G. P. Sachdev. 1994. A novel human airway mucin cDNA encodes a protein with unique tandem-repeat organization. Biochem. J. 300: 295-298 .
7. D'Cruz, O. J., T S. Dunn, P. Pichan, G. G. Hass Jr., and G. P. Sachdev. 1996. Antigenic cross-reactivity of human tracheal mucin with human sperm and trophoblasts correlates with the expression of mucin 8 gene messenger ribonucleic acid in reproductive tract tissues. Fertil. Steril. 66: 316-326 [Medline].
8. Pigny, P., V. Guyonnet-Duperat, A. S. Hill, W. S. Pratt, S. Galiegue-Zouitina, M. C. d'Hooge, A. Laine, I. Van-Seuningen, P. Degand, J. R. Gum, Y. S. Kim, D. M. Swallow, J. P. Aubert, and N. Porchet. 1996. Human mucin genes assigned to 11p15.5: identification and organization of a cluster of genes. Genomics 38: 340-352 [Medline].
9. Gambús, G., C. De Bolòs, D. Andreu, C. Francí, G. Egea, and F. X. Real. 1993. Detection of the MUC2 apomucin tandem repeat with a mouse monoclonal antibody. Gastroenterology 104: 93-102 [Medline].
10.
Ho, S. B.,
L. L. Shekels,
N. W. Toribara,
Y. S. Kim,
J. C. Lyftogt,
D. L. Cherwitz, and
G. A. Niehans.
1995.
Mucin gene expression in normal, preneoplastic, and neoplastic human gastric epithelium.
Cancer Res.
55:
2681-2690
11. Carrato, C., C. Balagué, C. De Bolòs, E. González, G. Gambús, J. Planas, J. M. Perini, A. Andreu, and F. X. Real. 1994. Differential apomucin expression in normal and neoplastic human gastrointestinal tissues. Gastroenterology 107: 160-172 [Medline].
12. Balagué, C., G. Gambús, C. Carrato, N. Porchet, J. P. Aubert, Y. S. Kim, and F. X. Real. 1995. Altered expression of MUC2, MUC4, and MUC5 mucin genes in pancreas cancer cell lines and tissues. Gastroenterology 106: 1054-1061 [Medline].
13. De Bolós, C., M. Gumà, C. Barranco, M. Garrido, Y. S. Kim, and F. X. Real. 1998. MUC6 expression in breast tissues and cultured cell lines: abnormal expression in tumors and regulation by steroid hormones. Int. J. Cancer 77: 193-199 [Medline].
14.
Li, J. D.,
W. Feng,
M. Gallup,
J. H. Kim,
J. Gum,
Y. S. Kim, and
C. Basbaum.
1998.
Activation of NF-B via a Src-dependent Ras-MAPK-pp90rsk pathway is required for Pseudomonas aeruginosa-induced mucin overproduction in epithelial cells.
Proc. Natl. Acad. Sci. USA
95:
5718-5723
15.
Li, J. D.,
M. Gallup,
N. Fan,
D. E. Szymkowski, and
C. B. Basbaum.
1998.
Cloning the amino-terminal and 5'-flanking region of the human
MUC5AC mucin gene and the transcriptional up-regulation by bacterial
exoproducts.
J. Biol. Chem.
273:
6812-6820
16.
Clausen, H., and
E. P. Bennett.
1996.
A family of UDP-GalNAc: polypeptide N-acetylgalactosaminyl-transferases control the initiation of mucin-type O-linked glycosylation.
Glycobiology
6:
635-646
17. Kim, K. C., K. McCracken, B. C. Lee, C. Y. Shin, M. J. Jo, C. J. Lee, and K. H. Ko. 1997. Airway goblet cell mucin: its structure and regulation of secretion. Eur. Respir. J. 10: 2644-2649 [Abstract].
18.
Xing, P. X.,
K. Reynolds,
J. J. Tjandra,
X. L. Tang, and
I. F. C. McKenzie.
1990.
Synthetic peptides reactive with anti-human milk fat globule membrane monoclonal antibodies.
Cancer Res.
50:
89-98
19. Klomp, L. W. J., L. Van Rens, and G. L. Strous. 1995. Cloning and analysis of human gastric cDNA reveals two types of conserved cysteine-rich domains. Biochem. J. 308: 831-838 .
20.
López-Ferrer, A.,
C. De Bolòs,
C. Barranco,
M. Garrido,
J. Isern,
I. Carlsted,
C. A. Reis,
J. Torrado, and
F. X. Real.
2000.
Role of fucosyltransferases in the association between apomucin and Lewis antigen expression
in normal and malignant gastric epithelium.
Gut
47:
349-356
21. De Bolòs, C., M. Garrido, and F. X. Real. 1995. MUC6 apomucin shows a distinct normal tissue distribution that correlates with Lewis antigen expression in the human stomach. Gastroenterology 109: 723-734 [Medline].
22. Van-Klinken, B. J. W., J. Dekker, H. A. Büller, C. De Bolòs, and A. W. C. Einhard. 1997. Biosynthesis of mucins (MUC2-6) along the longitudinal axis of the human gastrointestinal tract. Am. J. Physiol. 273: 296-302 .
23.
Bernacki, S. H.,
A. L. Nelson,
L. Abdullah,
J. K. Sheehan,
A. Harris,
C. D. Davis, and
S. H. Randell.
1999.
Mucin gene expression during differentiation of human airway epithelia in vitro.
Am. J. Respir. Cell Mol. Biol.
20:
595-604
24. Audié, J. P., A. Janin, N. Porchet, M. C. Copin, P. Degand, B. Gosselin, and J. P. Aubert. 1993. Expression of human mucin genes in respiratory, digestive and reproductive tracts ascertained by in situ hybridization. J. Histochem. Cytochem. 41: 1479-1485 [Abstract].
25. Voynow, J. A., and M. C. Rose. 1994. Quantitation of mucin RNA in respiratory and intestinal epithelial cells. Am. J. Respir. Cell Mol. Biol. 11: 742-758 [Abstract].
26.
Sharma, P.,
L. Dudus,
P. A. Nielsen,
H. Clausen,
J. R. Yankaskas,
M. A. Hollingsworth, and
J. F. Engelhardt.
1998.
MUC5B and MUC7 are differentially expressed in mucous and serous cells of submucosal glands in human bronchial airways.
Am. J. Respir. Cell Mol. Biol.
19:
30-37
27.
Socci, G.,
M. Miozzo,
U. Pastorino,
S. Pilotti,
R. Donghi,
M. Giarola,
L. De
Gregorio,
G. Mananti,
P. Radice,
F. Minoletti,
G. Della,
Porta, and
M. Pierotti.
1995.
Genetic evidence for an independent origin of multiple preneoplastic and neoplastic lung lesions.
Cancer Res.
55:
135-140
28. Zotter, S., P. C. Hageman, A. Lossnitzer, W. J. Mooi, and J. Hilgers. 1988. Tissue and tumor distribution of human polymorphic epithelial mucin. Cancer Rev. 11-12:55-101.
29. Carter, D.. 1983. Small-cell carcinoma of the lung. Am. J. Surg. Pathol. 7: 787-795 [Medline].
30. McDowell, E. M., and B. F. Trump. 1983. Histogenesis of preneoplastic and neoplastic lesions in tracheobronchial epithelium. Surv. Synth. Pathol. Res. 2: 235-279 .
31.
Socci, G.,
U. Pastorino,
L. Moiraghi,
E. Tagliablue,
F. Pezella,
C. Ghirelli,
S. Tornielli,
L. Sard,
K. Huebner,
M. A. Pierotti,
C. M. Croce, and
S. Pilotti.
1998.
Loss of FHIT function in lung cancer and preinvasive bronchial lesions.
Cancer Res.
58:
5032-5037
32.
Sugio, F.,
Y. Kishimoto,
A. K. Virmani,
J. Y. Hung, and
A. F. Gazdar.
1994.
K-ras mutations are a relatively late event in the pathogenesis of lung cancer.
Cancer Res.
54:
5811-5815
33.
Schneider, S. S.,
C. Schik,
K. E. Fish,
E. Miller,
J. C. Pene,
S. D. Treter,
S. M. Hui, and
G. A. Silverman.
1995.
A serine proteinase inhibitor locus
at 18q21.3 contains a tandem duplication of the human squamous cell carcinoma antigen gene.
Proc. Natl. Acad. Sci. USA
92:
3147-3151
34. Nguyen, P. L., G. A. Niehans, D. L. Cherwtiz, Y. D. Kim, and S. B. Ho. 1996. Membrane-bound (MUC1) and secretory (MUC2, MUC3, and MUC4) mucin gene expression in human lung cancer. Tumor Biol. 17: 176-192 .
35. Seregni, E., C. Botti, C. Lombardo, A. Cantoni, A. Bogni, I. Cataldo, and E. Bombardieri. 1996. Pattern of mucin gene expression in normal and neoplastic lung tissues. Anticancer Res. 16: 2209-2213 [Medline].
36. Yu, C.-J., P.-C. Yang, C.-T. Shun, Y.-C. Lee, S.-H. Kuo, and K.-T. Luh. 1996. Overexpression of MUC5 genes is associated with early post-operative metastasis in non-small cell lung carcinoma. Int. J. Cancer 69: 457-467 [Medline].
37.
Yu, C.-J.,
J.-Y. Shew,
C.-T. Shun,
H.-T. Lin,
S.-H. Kuo,
K.-T. Luh, and
P.-C. Yang.
1998.
Quantitave analysis of mRNA encoding MUC1, MUC2, and
MUC5AC genes: a correlation between specific mucin gene expression
and sialomucin expression in non-small cell lung cancer.
Am. J. Respir.
Cell Mol. Biol.
18:
643-652
38. Copin, M. C., L. Devisme, M. P. Buisine, C. H. Marquette, A. Wurtz, J. P. Aubert, B. Gosselin, and N. Porchet. 2000. From normal respiratory mucosa to epidermoid carcinoma: expression of human mucin genes. Int. J. Cancer 86: 162-168 [Medline].
39.
Hennebicq-Reig, S.,
T. Lesuffleur,
C. Capon,
C. De Bolós,
I. Kim,
O. Moreau,
C. Richet,
B. Hémon,
M.-A. Recchi,
E. Maës,
J. P. Aubert,
F. X. Real,
A. Zweibaum,
P. Delannoy,
P. Degand, and
G. Huet.
1998.
Permanent exposure of mucin-secreting HT-29 cells to benzyl-N-acetyl--D-galactosaminide induces abnormal O-glycosylation of mucins and inhibits
constitutive and stimulated MUC5AC secretion.
Biochem. J.
334:
283-285
.
40. Barasch, J., B. Kiss, A. Prince, L. Saiman, D. Gruenert, and Q. Al-Awqati. 1991. Defective acidification of intracellular organelles in cystic fibrosis. Nature 352: 70-73 [Medline].
41. Zhang, Y., B. Doranz, J. R. Yankasas, and J. F. Engelhardt. 1995. Genotypic analysis of respiratory mucus sulfation defects in cystic fibrosis. J. Clin. Invest. 96: 2997-3004 .
42. Zhang, S., H. S. Zhang, C. Cordon-Cardo, V. E. Reuter, A. K. Singhal, K. O. Lloyd, and P. O. Livignston. 1999. Selection of tumor antigens as targets for immune attack using immunohistochemistry: II. Blood group-related antigens. Int. J. Cancer 73: 50-56 .
43. Toma, V., T. Sata, P. Vogt, P. Komminoth, P. U. Heitz, and J. Roth. 1999. Differentiation-related expression of the Thomsen-Friedenreich glycotope in developing human lung and in lung carcinoma. Cancer 85: 2151-2159 [Medline].
44.
Molinolo, A.,
J. F. Simpson,
A. Thor, and
J. Schlom.
1990.
Enhanced tumor
binding using immunohistochemical analyzes by second generation anti-tumor-associated glycoprotein 72 monoclonal antibodies versus monoclonal antibody B72.3 in human tissue.
Cancer Res.
50:
1291-1298
45. Zenita, K., Y. Kirihata, A. Kitahara, K. Shigeta, K. Higuchi, K. Hirashima, T. Murachi, M. Miyake, T. Takeka, and R. Kannagi. 1988. Fucosylated type 2 chain polylactosamine antigens in human lung cancer. Int. J. Cancer 41: 344-349 [Medline].
46. Kudo, T., Y. Ikehara, A. Togayachi, K. Morozumi, M. Watanabe, M. Nakamura, S. Nishihara, and H. Narimatsu. 1998. Up-regulation of a set of glycosyltransferase genes in human colorectal cancer. Lab. Invest. 78: 797-811 [Medline].
47.
Mandel, U.,
H. Hassan,
M. H. Therkildsen,
J. Rygaard,
M. H. Jakobsen,
B. R. Juhl,
E. Dabelsteen, and
H. Clausen.
1999.
Expression of polypeptide GalNAc-transferases in stratified epithelia and squamous cell carcinomas: immunohistological evaluation using monoclonal antibodies to three
members of the GalNAc-transferase family.
Glycobiology
9:
43-52
48. Egea, G., C. Francí, G. Gambús, T. Lesuffleur, A. Zweibaum, and F. X. Real. 1993. cis-Golgi resident proteins and O-glycans are abnormally compartmentalized in the RER of colon cancer cells. J. Cell Sci. 105: 819-830 [Abstract].
This article has been cited by other articles:
 |
|
 |
|
  S. Ocak, M. L. Sos, R. K. Thomas, and P. P. Massion High-throughput molecular analysis in lung cancer: insights into biology and potential clinical applications Eur. Respir. J., August 1, 2009; 34(2): 489 - 506. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. H. SCHMIDT, S. BIESTERFELD, A. KUMMEL, A. FALDUM, M. SEBASTIAN, C. TAUBE, R. BUHL, and R. WIEWRODT Tissue Microarrays are Reliable Tools for the Clinicopathological Characterization of Lung Cancer Tissue Anticancer Res, January 1, 2009; 29(1): 201 - 209. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. R. White, B. M. Fischer, B. A. Marroquin, and R. Stern Interleukin-1{beta} mediates human airway epithelial cell migration via NF-{kappa}B Am J Physiol Lung Cell Mol Physiol, December 1, 2008; 295(6): L1018 - L1027. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Chaturvedi, A. P. Singh, and S. K. Batra Structure, evolution, and biology of the MUC4 mucin FASEB J, April 1, 2008; 22(4): 966 - 981. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. W. Williams, A. Sharafkhaneh, V. Kim, B. F. Dickey, and C. M. Evans Airway Mucus: From Production to Secretion Am. J. Respir. Cell Mol. Biol., May 1, 2006; 34(5): 527 - 536. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. C. Rose and J. A. Voynow Respiratory Tract Mucin Genes and Mucin Glycoproteins in Health and Disease Physiol Rev, January 1, 2006; 86(1): 245 - 278. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Sriuranpong, A. Mutirangura, J. W. Gillespie, V. Patel, P. Amornphimoltham, A. A. Molinolo, V. Kerekhanjanarong, S. Supanakorn, P. Supiyaphun, S. Rangdaeng, et al. Global Gene Expression Profile of Nasopharyngeal Carcinoma by Laser Capture Microdissection and Complementary DNA Microarrays Clin. Cancer Res., August 1, 2004; 10(15): 4944 - 4958. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Nishiumi, Y. Abe, Y. Inoue, H. Hatanaka, K.-i. Inada, H. Kijima, H. Yamazaki, M. Tatematsu, Y. Ueyama, M. Iwasaki, et al. Use of 11p15 Mucins as Prognostic Factors in Small Adenocarcinoma of the Lung Clin. Cancer Res., November 15, 2003; 9(15): 5616 - 5619. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. J. Korst and R. G. Crystal Active, specific immunotherapy for lung cancer: hurdles and strategies using genetic modification Ann. Thorac. Surg., October 1, 2003; 76(4): 1319 - 1326. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. D. Leikauf, M. T. Borchers, D. R. Prows, and L. G. Simpson Mucin Apoprotein Expression in COPD* Chest, May 1, 2002; 121 (2009): 166S - 182S. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-J. YU, J.-Y. SHEW, Y.-S. LIAW, S.-H. KUO, K.-T. LUH, and P.-C. YANG Application of Mucin Quantitative Competitive Reverse Transcription Polymerase Chain Reaction in Assisting the Diagnosis of Malignant Pleural Effusion Am. J. Respir. Crit. Care Med., October 1, 2001; 164(7): 1312 - 1318. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Proc. Am. Thorac. Soc. | Am. J. Respir. Crit. Care Med. |