metallothionein and Esophageal-Neoplasms

This review summarizes and evaluates the literature regarding the biomarkers for predicting the response and/or prognosis of esophageal squamous cell carcinoma (ESCC) patients treated with neoadjuvant chemoradiation therapy (CRT). There are seven categories of molecules known to correlate with the response and/or prognosis: tumor suppressors (p53, p21), cell cycle regulators (Cyclin D1, CDC25B, 14-3-3sigma), DNA repair molecules (p53R2, ERCC1), drug resistance proteins [metallothionein (MT)], angiogenic factors (VEGF), molecules involved in cell proliferation/invasion/metastasis (Ki-67, COX-2) and hedgehog signaling molecules (Gli-1). Of the above molecules, the tumor suppressor p53 is expected to be a representative biomarker for predicting the response and prognosis. The cell cycle markers CDC25B and 14-3-3sigma have potential as response biomarkers independent of the p53 status. The DNA repair markers, p53R2 or ERCC1, angiogenic molecule (VEGF), and hedgehog signaling pathway factor Gli-1 also have potential to predict the response and prognosis of ESCC. However, there are still many unanswered questions with regard to predicting the clinical effects of neoadjuvant CRT.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Cycle Proteins; Chemoradiotherapy, Adjuvant; Cyclooxygenase 2; DNA-Binding Proteins; Endonucleases; Esophageal Neoplasms; Forecasting; Humans; Ki-67 Antigen; Meta-Analysis as Topic; Metallothionein; Neoadjuvant Therapy; Prognosis; Ribonucleotide Reductases; Tumor Suppressor Protein p53; Vascular Endothelial Growth Factor A

Metallothionein (

Topics: Biomarkers, Tumor; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Esophageal Neoplasms; Female; Humans; Male; Metallothionein; Middle Aged; Phosphatidylinositol 3-Kinases; Superoxide Dismutase-1

To explore the anti-tumor effects of esophageal cancer-related gene 2 (ECRG2) in combination with cisplatin (DDP) in DDP-resistant esophageal cancer cells (EC9706/DDP).. A drug-resistant cell model was established, with EC9706/DDP cells being treated with ECRG2 and/or DDP. Cell viability was examined by MTT assay. The rate of cell apoptosis was determined by flow cytometry. The mRNA expression levels of proliferating cell nuclear antigen (PCNA), metallothionein (MT), and p53 were determined by RT-PCR and PCNA, while MT and p53 protein expression levels were determined by western blotting.. The anti-proliferative effect of ECRG2 in combination with DDP was superior when compared to ECRG2 or DDP alone. The inhibition rate for the combination reached its peak (51.33%) at 96 h. The early apoptotic rates of the control, ECRG2 alone, DDP alone, and ECRG2 plus DDP groups were 5.71% ± 0.27%, 12.68% ± 0.61%, 14.15% ± 0.87%, and 27.96% ± 0.36%, respectively. Although all treatment groups were significantly different from the control group (

Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cisplatin; Down-Regulation; Drug Resistance, Neoplasm; Drug Therapy, Combination; Esophageal Neoplasms; Flow Cytometry; Humans; Metallothionein; Proliferating Cell Nuclear Antigen; Proteinase Inhibitory Proteins, Secretory; Recombinant Proteins; RNA, Messenger; Serine Peptidase Inhibitors, Kazal Type; Transcriptional Activation; Tumor Suppressor Protein p53; Up-Regulation

Metallothionein (MT)-3 has cell growth inhibitory activity, and is the only currently known MT subtype with unique physiological functions. The expression levels of MT-1E, a subtype of MT-1, were positively correlated with the degree of esophageal cancer malignancy. The present study aimed to investigate the effects of MT-3 and MT-1E gene transfection on the proliferation, cell cycle, and apoptosis of esophageal cancer cells. The cationic liposome method was used to transfect the esophageal cancer strains Eca-109 and TE13. Reverse transcription-polymerase chain reaction was used to detect target gene expression, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction was applied to detect cell proliferation, and flow cytometry was used for cell cycle and apoptosis detection. Esophageal cancer cells with MT-3 and MT-1E gene transfection showed high expression of the foreign target gene and mRNA. Cells with MT-3 gene transfection showed markedly inhibited proliferation (P < 0.05), a significantly higher proportion of cells in the G0/G1 phase (P < 0.05), a significantly lower proportion of cells in the S phase (P < 0.05), and a significantly increased apoptosis rate (P < 0.05). Cells with MT-1E gene transfection did not show significant changes in proliferation, cell cycle, or apoptosis rate (P > 0.05). Therefore, the upregulation of MT-3 gene expression can inhibit esophageal cancer cell proliferation and induce apoptosis, which may be achieved by blocking the tumor cell growth cycle, whereas effects of the MT-1E gene on the proliferation of esophageal cancer cells were not evident.

Topics: Apoptosis; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Esophageal Neoplasms; Gene Expression; Humans; Metallothionein; Metallothionein 3; Nerve Tissue Proteins; Transfection

The incidence of esophageal squamous cell carcinoma (ESCC) is very high in northern China. This cancer has a very poor prognosis, mostly because it is usually diagnosed at a late stage. Detection at an earlier stage can dramatically improve prognosis. Microscopic evaluation of esophageal balloon cytology (EBC) specimens has been the most common method for early detection of ESCC, but this technique is limited by low sensitivity and specificity. The use of molecular markers may improve these screening characteristics. This study evaluates whether measurement of gene methylation in EBC specimens may have utility for the detection of esophageal squamous dysplasia and early ESCC. We evaluated the presence of methylation in eight genes shown to be methylated in ESCC in previous studies in EBC specimens from 147 patients with endoscopic biopsy diagnoses ranging from normal mucosa to severe squamous dysplasia. Methylation status was determined using quantitative methylation-specific PCR techniques. The sensitivity and specificity of methylation of each individual gene and of combinations of these genes to detect biopsyproven high-grade (moderate or severe) squamous dysplasia were determined. For individual genes, the sensitivities ranged from 9% to 34% and the specificities ranged from 77% to 99%. Using a panel of four genes (AHRR, p16INK4a, MT1G, and CLDN3) resulted in sensitivity and specificity of 50% and 68%, respectively. This study suggests that evaluation of gene methylation in EBC samples may have utility for early detection of esophageal squamous dysplasia and early ESCC; however, identification of more sensitive methylation markers will be required for development of a clinically useful screening test.

Topics: Adult; Aged; Barrett Esophagus; Basic Helix-Loop-Helix Transcription Factors; Biomarkers, Tumor; Carcinoma, Squamous Cell; Claudin-3; DNA Methylation; Early Detection of Cancer; Esophageal Neoplasms; Esophagoscopy; Female; Genes, p16; Humans; Male; Membrane Proteins; Metallothionein; Middle Aged; Precancerous Conditions; Promoter Regions, Genetic; Repressor Proteins; Sensitivity and Specificity

Metallothionein (MT)-3 can inhibit cell growth. Up to now, it is the only MT subtype with specific physiological function. This study was to investigate the expression of MT-3 in esophageal cancer and its correlations to the expression of MT-1 and MT-2.. The expression of MT-1, -2 and -3 in 76 specimens of esophageal cancer and 10 specimens of esophageal diverticulum were detected by SP immunohistochemistry. The correlations of MT-3 expression to MT-1 and MT-2 expression were analyzed.. The expression level of MT-3 was significantly higher in esophageal cancer than in para-cancer normal tissues and benign esophageal lesions (Chi2=74.696, P<0.001). In tumor tissues, MT-3 was mainly expressed in cytoplasm. The expression level of MT-3 was related with differentiation (P=0.004), TNM stage (P=0.033), lymph node metastasis (P=0.017) and survival time (P=0.035), and negatively correlated to the expression levels of MT-1 and MT-2 (r=0.247, P=0.032).. MT-3 is overexpressed in esophageal cancer. Its expression is closely related with the differentiation, development and prognosis of esophageal cancer, and its roles are obviously different from those of MT-1 and MT-2.

Topics: Adult; Aged; Esophageal Neoplasms; Esophagus; Female; Humans; Immunohistochemistry; Male; Metallothionein; Metallothionein 3; Middle Aged; Nerve Tissue Proteins

Esophageal squamous cell carcinoma (ESCC) in the Indian population is associated with poor nutritional status, low socioeconomic conditions, bidi smoking and consumption of smokeless tobacco products, besides alcohol drinking and cigarette smoking. To determine the impact of these risk factors on molecular pathogenesis of ESCC, we determined global gene expression profiles of 7 paired samples of ESCC and histologically confirmed nonmalignant esophageal tissues using 19.1K cDNA microarrays. The most salient finding was identification of 19 differentially expressed genes encoding zinc binding or modulating proteins associated with transcriptional regulation, ubiquitin-protein degradation and maintenance of zinc homeostasis. Validation of differential expression of a subset of genes by real-time quantitative RT-PCR (real-time QRT-PCR) in clinical specimens of ESCC, esophageal dysplasia and histologically nonmalignant esophageal tissues and immunohistochemical analysis using tissue microarrays confirmed the microarray data and demonstrated upregulation of zinc finger proteins, cellular modulator of immune recognition (c-MIR), snail homolog 2 (SLUG), zinc transporter, ZnT7 and downregulation of zinc metabolizing protein, metallothionein MT1G. We also observed upregulation of mitogen activated protein kinase kinase kinase 3 (MAP3K3/MEKK3), a kinase anchor protein 13 (AKAP13) and transglutaminase2 (TG2). Interestingly, we found upregulation of ZnT7 transcripts in ESCC cells (TE13) grown in zinc deficient condition. In conclusion, our data suggest deregulation of genes associated with zinc homeostasis in ESCC.

Topics: Carcinoma, Squamous Cell; Cation Transport Proteins; Esophageal Neoplasms; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Homeostasis; Humans; Immunohistochemistry; Male; Metallothionein; Neoplasm Proteins; Oligonucleotide Array Sequence Analysis; RNA, Neoplasm; Tumor Cells, Cultured; Zinc; Zinc Fingers

Upper aerodigestive tract (UADT) cancer, including oral and esophageal cancer, is an important cause of cancer deaths worldwide. Patients with UADT cancer are frequently zinc deficient (ZD) and show a loss of function of the pivotal tumor suppressor gene p53. The present study examined whether zinc deficiency in collaboration with p53 insufficiency (p53+/-) promotes lingual and esophageal tumorigenesis in mice exposed to low doses of the carcinogen 4-nitroquinoline 1-oxide. In wild-type mice, ZD significantly increased the incidence of lingual and esophageal tumors from 0% in zinc sufficient (ZS) ZS:p53+/+ mice to approximately 40%. On the p53+/- background, ZD:p53+/- mice had significantly greater tumor incidence and multiplicity than ZS:p53+/- and ZD:p53+/+ mice, with a high frequency of progression to malignancy. Sixty-nine and 31% of ZD:p53+/- lingual and esophageal tumors, respectively, were squamous cell carcinoma versus 19 and 0% of ZS:p53+/- tumors (tongue, P = 0.003; esophagus, P = 0.005). Immunohistochemical analysis revealed that the increased cellular proliferation observed in preneoplastic lingual and esophageal lesions, as well as invasive carcinomas, was accompanied by overexpression of cytokeratin 14, cyclooxygenase-2 and metallothionein. In summary, a new UADT cancer model is developed in ZD:p53+/- mouse that recapitulates aspects of the human cancer and provides opportunities to probe the genetic changes intrinsic to UADT carcinogenesis and to test strategies for prevention and reversal of this deadly cancer.

Topics: 4-Nitroquinoline-1-oxide; Animals; Biomarkers, Tumor; Carcinogens; Cyclooxygenase 2; Disease Models, Animal; Esophageal Neoplasms; Female; Gene Expression; Genetic Predisposition to Disease; Immunohistochemistry; Keratin-14; Keratins; Male; Metallothionein; Mice; Mice, Mutant Strains; Precancerous Conditions; Tongue Neoplasms; Tumor Suppressor Protein p53; Zinc

Esophageal squamous cell carcinoma (ESCC) is a common cancer with a very poor prognosis. New methods are needed to screen high-risk populations and identify curable tumors and precursor lesions early. Molecular markers may be useful in such screening efforts. This study was designed to determine the prevalence of p16, MGMT, RARbeta2, CLDN3, CRBP and MT1G gene methylation in patients with ESCC to evaluate the variation of gene methylation across a spectrum of preneoplastic lesions, and assess the feasibility of using gene methylation in a primary screening test utilizing frozen esophageal cells collected by balloon cytology samplers. Samples were obtained from high-risk subjects from north central China. These samples included 11 foci of histologically normal mucosa, 8 foci of low-grade squamous dysplasia, 7 foci of high-grade squamous dysplasia, and 13 foci of ESCC from 6 fully embedded resection specimens; endoscopic biopsies from 6 individuals with no histological evidence of disease; and frozen esophageal balloon samples from 12 asymptomatic subjects. Promoter CpG site-specific hypermethylation status was determined for each gene using real-time methylation-specific PCR (qMS-PCR) based on Taqman chemistry. Of the 6 ESCC patients, 5 showed methylation of at least one gene. For most genes, methylation occurred with increasing frequency during neoplastic progression, with the largest increase found between low- and high-grade dysplasia. There was considerable variation in methylation patterns among different foci of the same histological grade, even within individual patients, but 16/20 (80%) of high-grade dysplastic and cancer foci had >or= 2 methylated genes, while 17/19 (89%) of normal and low-grade dysplastic foci had <2 methylated genes. These genes were rarely methylated in histologically normal mucosa from patients with or without ESCC. Gene methylation was common and easily detectable in the frozen esophageal cells collected by balloon cytology samplers. Our data suggest that methylation of p16, MGMT, RARbeta2, CLDN3, CRBP, and MT1G is common in the esophageal mucosa of patients with ESCC in this high-risk population, and tends to increase in prevalence in foci with increasing histological severity of disease. Methylation data from panels of genes may be able to identify patients with high-grade lesions. Balloon cytology may be able to screen the length of the esophagus effectively for a subset of cells with abnormal methylation, and may be useful

Topics: Adult; Aged; Carcinoma, Squamous Cell; Claudin-3; DNA Methylation; DNA Modification Methylases; DNA Repair Enzymes; Esophageal Neoplasms; Female; Genes, p16; Humans; Male; Membrane Proteins; Metallothionein; Middle Aged; Precancerous Conditions; Receptors, Retinoic Acid; Retinol-Binding Proteins; Retinol-Binding Proteins, Cellular; Tumor Suppressor Protein p14ARF; Tumor Suppressor Proteins

Zinc deficiency in rats enhances esophageal cell proliferation, causes alteration in gene expression, and promotes esophageal carcinogenesis. Zinc replenishment rapidly induces apoptosis in the esophageal epithelium thereby reversing cell proliferation and carcinogenesis. To identify zinc-responsive genes responsible for these divergent effects, we did oligonucleotide array-based gene expression profiling analyses in the precancerous zinc-deficient esophagus and in zinc-replenished esophagi after treatment with intragastric zinc compared with zinc-sufficient esophagi. Thirty-three genes (21 up-regulated and 12 down-regulated) showed a > or = 2-fold change in expression in the hyperplastic zinc-deficient versus zinc-sufficient esophageal epithelia. Expression of genes involved in cell division, survival, adhesion, and tumorigenesis were markedly changed. The zinc-sensitive gene metallothionein-1 (MT-1 was up-regulated 7-fold, the opposite of results for small intestine and liver under zinc-deficient conditions. Keratin 14 (KRT14, a biomarker in esophageal tumorigenesis), carbonic anhydrase II (CAII, a regulator of acid-base homeostasis), and cyclin B were up-regulated >4-fold. Immunohistochemistry showed that metallothionein and keratin 14 proteins were overexpressed in zinc-deficient esophagus, as well as in lingual and esophageal squamous cell carcinoma from carcinogen-treated rats, emphasizing their roles in carcinogenesis. Calponin 1 (CNN1, an actin cross-linking regulator) was down-regulated 0.2-fold. Within hours after oral zinc treatment, the abnormal expression of 29 of 33 genes returned to near zinc-sufficient levels, accompanied by reversal of the precancerous phenotype. Thus, we have identified new molecular markers in precancerous esophagus and showed their restoration by zinc replenishment, providing insights into the interaction between zinc and gene expression in esophageal cancer development and prevention.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinoma, Squamous Cell; Esophageal Neoplasms; Gene Expression Regulation, Neoplastic; Immunohistochemistry; Keratin-14; Keratins; Male; Metallothionein; Precancerous Conditions; Rats; Reverse Transcriptase Polymerase Chain Reaction; Up-Regulation; Zinc

Esophageal squamous cell carcinoma is generally sensitive to chemoradiotherapy (CRT), but some cases are not. Using a retrospective analysis, we aimed to identify the predictors of the response by esophageal squamous cell carcinoma to definitive CRT.. The intensities of expression of p53, Ki67, Bcl-2, Bax, cyclin D1, VEGF, CDC25B, and metallothionein (MT) were evaluated immunohistochemically in the biopsy specimens obtained before CRT, and the intensities of their expression were tested for correlations with the clinical effects of CRT.. The esophageal squamous cell carcinomas with negative p53, positive CDC25B, and negative MT expression were found to be significantly more sensitive to CRT. In addition, p53 positivity and CDC25B positivity respond well to CRT.. Esophageal squamous cell carcinomas with negative p53, positive CDC25B, and negative MT expressions respond well to CRT. Even with p53 positivity, if with CDC25B positivity, CRT can be expected.

Topics: Adult; Aged; Antineoplastic Agents; Biomarkers, Tumor; Carcinoma, Squamous Cell; cdc25 Phosphatases; Cell Cycle Proteins; Esophageal Neoplasms; Female; Humans; Male; Metallothionein; Middle Aged; Retrospective Studies; Tumor Suppressor Protein p53

Several studies have shown that macroscopically infiltrative type of esophageal carcinoma generally has a poorer local response to radiation therapy than that of localized type. The aim of this study was to determine the role of metallothionein (MT) as a radioprotective agent in the difference of clinical radiosensitivities in esophageal carcinoma. A total of 45 surgically resected esophageal carcinoma tissues [20 macroscopically localized type without preoperative treatment (PT), 20 macroscopically infiltrative type without PT and 5 macroscopically infiltrative type with PT] were stained for MT by immunohistochemistry and were analyzed. MT expression level of macroscopically localized type of esophageal carcinoma was significantly higher than that of infiltrative type without PT (P=0.026) and was significantly higher than that of infiltrative type with PT (P=0.024). No significant difference was found between MT expression levels in infiltrative type of esophageal carcinoma without PT and that with PT. The results of this study suggest that there is less possibility that clinical radioresistance of the tumor is due to MT expression in esophageal carcinoma and also suggest that there is less possibility that MT synthesis is induced by fractionated irradiation of a moderate dose or by an anti-cancer agent during a course of treatment.

Topics: Aged; Esophageal Neoplasms; Female; Humans; Immunohistochemistry; Male; Metallothionein; Middle Aged; Radiation Tolerance

Barrett's esophagus is the transformation of normal esophageal squamous epithelium to specialized intestinal metaplasia (SIM). Among the Barrett's specialized cells, those that can develop protective mechanisms against apoptosis may have potential to become malignant. Studies have shown that overexpression of metallothionein (MT), low molecular protein that protects cells from apoptotic stimuli, appears to be associated with more advanced, highly malignant tumors. We thus investigated the relationship between MT expression and apoptosis in different stages of Barrett's carcinogenesis. Terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling and immunohistochemical dual-staining assay were performed in human biopsy samples of normal, SIM, dysplasia, and adenocarcinoma. Apoptotic index and MT expression were quantified by using an image system to analyze the converted digital data. A negative correlation between MT expression and apoptotic index was found. MT expression was significantly increased along with the histologic progression towards adenocarcinoma. This study thus suggests that MT may contribute to cytoprotection, thereby inhibiting apoptosis and leading to carcinogenesis of Barrett's esophageal cells.

Topics: Apoptosis; Barrett Esophagus; Disease Progression; Esophageal Neoplasms; Humans; Image Processing, Computer-Assisted; Immunohistochemistry; Metallothionein

Esophageal cancer related gene 2 (ECRG2) is a novel candidate of the tumor suppressor gene identified from human esophagus. To study the biological role of the ECRG2 gene, we performed a GAL4-based yeast two-hybrid screening of a human fetal liver cDNA library. Using the ECRG2 cDNA as bait, we identified nine putative clones as associated proteins. The interaction of ECRG2 and metallothionein 2A (MT2A) was confirmed by glutathione S-transferase pull-down assays in vitro and co-immunoprecipitation experiments in vivo. ECRG2 co-localized with MT2A mostly to nuclei and slightly to cytoplasm, as shown by confocal microscopy. Transfection of ECRG2 gene inhibited cell proliferation and induced apoptosis in esophageal cancer cells. In the co-transfection of ECRG2 and MT2A assays, cell proliferation was inhibited and apoptosis was slightly induced compared with control groups. When we used antisense MT2A to interdict the effect of MT2A, the inhibition of cell proliferation and induction of apoptosis were significantly enhanced. When we used antisense ECRG2 to interdict the effect of ECRG2 in the group of Bel7402 cells co-transfected with ECRG2 and MT2A, the inhibition of cell proliferation and induction of apoptosis disappeared. The results provide evidence for ECRG2 in esophageal cancer cells acting as a bifunctional protein associated with the regulation of cell proliferation and induction of apoptosis. ECRG2 might reduce the function of MT2A on the regulation of cell proliferation and induction of apoptosis. The physical interaction of ECRG2 and MT2A may play an important role in the carcinogenesis of esophageal cancer.

Topics: Apoptosis; Cell Cycle; Cell Line; Esophageal Neoplasms; Flow Cytometry; Genes, Reporter; Genes, Tumor Suppressor; Genome, Human; Humans; Metallothionein; Microscopy, Confocal; Molecular Sequence Data; Protein Binding; Proteinase Inhibitory Proteins, Secretory; Recombinant Fusion Proteins; Serine Peptidase Inhibitors, Kazal Type; Tumor Suppressor Proteins; Two-Hybrid System Techniques

The potential of the metal-binding protein, metallothionein, in assessing the progression of normal oesophagus through Barrett's to adenocarcinoma was investigated. Metallothionein was quantitatively determined in resected tissues from patients undergoing oesophagectomy for high grade dysplasia/adenocarcinoma and in biopsies from patients with Barrett's syndrome. In 10 cancer patients, metallothionein concentrations in adenocarcinoma were not significantly different from normal oesophagus, although six had elevated metallothionein concentrations in the metaplastic tissue bordering the adenocarcinoma. In 17 out of 20 non-cancer patients with Barrett's epithelium, metallothionein was significantly increased by 108% (P<0.004). There was no association between the metallothionein levels in Barrett's epithelium and the presence of inflammatory cells, metaplasia or dysplasia. Metallothionein is a marker of progression from normal to Barrett's epithelium but is not increased in oesophageal adenocarcinoma.

Topics: Adenocarcinoma; Adult; Aged; Barrett Esophagus; Biomarkers; Biomarkers, Tumor; Biopsy; Disease Progression; Esophageal Neoplasms; Esophagitis; Esophagus; Female; Gastroesophageal Reflux; Humans; Hyperplasia; Male; Metallothionein; Metaplasia; Middle Aged; Neoplasm Metastasis; Neoplasm Proteins; Precancerous Conditions

The sensitivity of cancer cells to chemotherapy and radiation therapy depends on various biological properties. This study investigated the expression of p53, CDC25B and metallothionein (MT), and evaluated their clinical significance in chemoradiation therapy (CRT) for oesophageal squamous cell carcinoma.. The expression of p53, CDC25B and MT was evaluated by immunohistochemistry using biopsy specimens taken before CRT for 77 patients with oesophageal squamous cell carcinoma, and correlated with the pathological effects of CRT and survival.. p53-positive tumours and MT-positive tumours had a poor response to CRT, whereas tumours with strong CDC25B expression were associated with a good response. When each patient was scored for the presence of the three biological factors, there was a strong correlation between the sensitivity score and the pathological effect of CRT (P < 0.001), and a (non-significant) difference in the 5-year survival rate between patients with a high score and those with a low score (67 versus 34 per cent respectively; P = 0.12).. The combined evaluation of p53, CDC25B and MT may help to identify patients with advanced oesophageal squamous cell carcinoma who will benefit from preoperative CRT.

Topics: Adult; Aged; Aged, 80 and over; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Squamous Cell; cdc25 Phosphatases; Cell Cycle Proteins; Cisplatin; Esophageal Neoplasms; Female; Fluorouracil; Genes, p53; Humans; Immunohistochemistry; Infusions, Intravenous; Male; Metallothionein; Middle Aged; Survival Analysis; Treatment Outcome

The purpose of this study was to define the prognostic value of a group of molecular tumor markers in a well-staged population of patients treated with trimodality therapy for esophageal cancer. The original pretreatment paraffin-embedded endoscopic esophageal tumor biopsy material was obtained from 118 patients treated with concurrent cisplatin + 5-fluorouracil (5-FU) + 45 Gy radiation followed by resection from 1986 until 1997 at the Duke University Comprehensive Cancer Center. Three markers of possible platinum chemotherapy association [metallothionein (MT), glutathione S-transferase-pi (GST-pi), P-glycoprotein (P-gp or multidrug resistance)] and one marker of possible 5-FU association [thymidylate synthase (TS)] were measured using immunohistochemistry. The median cancer-free survival was 25.0 months, with a significantly improved survival for the 38 patients who had a complete response (P < 0.001). High-level expression of GST-pi, P-gp, and TS were associated with a decreased survival. MT was not significant in this population. Multivariate analysis identified high-level expression in two of the platinum markers (GST-pi and P-gp) and the 5-FU marker TS as independent predictors of early recurrence and death. In conclusion, this investigation measured three possible markers associated with platinum and one possible marker associated with 5-FU in a cohort of esophageal cancer patients. Independent prognostic significance was observed, which suggests that it may be possible to predict which patients may benefit most from trimodality therapy. These data need to be reproduced in a prospective investigation.

Topics: Adenocarcinoma; Adult; Age Factors; Aged; Antineoplastic Combined Chemotherapy Protocols; ATP Binding Cassette Transporter, Subfamily B, Member 1; Biopsy; Carcinoma, Squamous Cell; Cisplatin; Disease-Free Survival; Esophageal Neoplasms; Fluorouracil; Glutathione S-Transferase pi; Glutathione Transferase; Humans; Immunohistochemistry; Isoenzymes; Metallothionein; Middle Aged; Multivariate Analysis; Prognosis; Proportional Hazards Models; Thymidylate Synthase; Time Factors; Treatment Outcome

Oxidative damage has long been related to carcinogenesis in human cancers and animal cancer models. Recently a rat esophageal adenocarcinoma (EAC) model was established in our laboratory by using esophagoduodenal anastomosis (EDA) plus iron supplementation. Our previous study suggested that iron supplementation enhanced inflammation and the production of reactive nitrogen species in the esophageal epithelium, which could contribute to esophageal adenocarcinogenesis. Here we further characterized oxidative damage in this model. We were particularly interested in how excess iron was deposited in the esophagus, and which cells were targeted by oxidative damage. Male Sprague-Dawley rats received iron supplementation (50 mg Fe/kg/month, i.p.) starting 4 weeks after EDA. The animals were killed at 11, 30 or 35 weeks after surgery. EAC appeared as early as week 11 after surgery, and increased over time, up to 60% at 35 weeks after surgery. All EACs were well-differentiated mucinous adenocarcinoma at the squamocolumnar junction. Iron deposition was found at the squamocolumnar junction and in the area with esophagitis. Esophageal iron overload could result from transient increase of blood iron after i.p. injection, and the overexpression of transferrin receptor in the premalignant columnar-lined esophagus (CLE) cells. Oxidative damage to DNA (8-hydroxy-2'-deoxyguanosine), protein (carbonyl content) and lipid (thiobarbituric acid reactive substance) in the esophagus was significantly higher than that of the non-operated control. CLE cells were believed to be the target cells of oxidative damage because they overexpressed heme oxygenase 1 and metallothionein, both known to be responsive to oxidative damage. We propose that oxidative damage plays an important role in the formation of EAC in the EDA model, and a similar situation may occur in humans with gastroesophageal reflux and iron over-nutrition.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Adenocarcinoma, Mucinous; Anastomosis, Surgical; Animals; Barrett Esophagus; Cocarcinogenesis; Deoxyguanosine; Disease Models, Animal; DNA Adducts; Duodenum; Epithelial Cells; Esophageal Neoplasms; Esophagitis; Esophagus; Gastroesophageal Reflux; Heme Oxygenase (Decyclizing); Humans; Iron; Isoenzymes; Male; Metallothionein; Oxidative Stress; Postoperative Complications; Precancerous Conditions; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Receptors, Transferrin; Thiobarbituric Acid Reactive Substances

Immunohistochemical staining for metallothionein (MT) and p53 was performed on biopsy specimens of 30 patients with esophageal squamous cell carcinoma who had received curative resection following preoperative chemoradiation. The pathologic response to chemoradiation was a partial response in 19 cases and no change was observed in 11 cases. In 16 cases with MT-positive tumor, 10 (62.5%) showed no change. In 14 cases with MT-negative tumor, 13 (92.8%) showed partial response. In 8 patients with negative staining for p53 and MT, 7 were responders, whereas in 9 patients with positive staining for p53 and MT, 6 were nonresponders. The pathologic response was significantly associated with the prognosis (p = 0. 0167). The survival rate of the responders was significantly better than that of the nonresponders. These findings suggest that MT might be a prognostic marker, and consequently we can select the patients who will benefit from preoperative chemoradiation.

Topics: Chemotherapy, Adjuvant; Esophageal Neoplasms; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Male; Metallothionein; Middle Aged; Prognosis; Radiotherapy, Adjuvant; Survival Analysis; Treatment Outcome; Tumor Suppressor Protein p53

The goal of this study is to clarify whether the expression of metallothionein (MT) could affect the prognosis and the metastatic potential of squamous cell carcinoma (SCC) of the oesophagus. In paraffin-embedded specimens resected from 57 patients, MT mRNA and protein expressions were detected by in situ hybridization and immunohistochemistry respectively. The expression of MT was evaluated in respect of clinicopathologic variables and patients' survival. MT mRNA expression was significantly associated with the proportion of lymph node metastasis (71% in MT mRNA-positive tumours vs 42% in MT mRNA-negative tumours; P = 0.0343) and that of distant metastasis (29% in MT mRNA-positive tumours vs 5% in MT mRNA-negative tumours; P = 0.0452). In respect of MT protein expression, the frequency of distant metastasis was more common in MT-positive tumours than in MT-negative tumours (30% in MT-positive tumours vs 8% in MT-negative tumours; P = 0.0446). The survival rate of the patients with MT protein-negative tumours was significantly better than that of the patients with MT protein-positive tumours (P = 0.0340). There was a positive correlation between the expression of MT protein and that of proliferating cell nuclear antigen (P = 0.0018). Therefore, we conclude that MT expression, both at the mRNA and protein levels, may be a potential marker predicting metastatic and proliferative activities of oesophageal SCC.

Topics: Aged; Base Sequence; Carcinoma, Squamous Cell; Cell Division; Esophageal Neoplasms; Female; Humans; Immunohistochemistry; In Situ Hybridization; Male; Metallothionein; Middle Aged; Molecular Sequence Data; Proliferating Cell Nuclear Antigen; RNA, Messenger