New Research on Esophageal Cancer / Antonio Carminati, editor.

Format:

Book

Contributor:

Carminati, Antonio, 1953- editor.

Series:

Horizons in cancer research.

Horizons in Cancer Research Series

Language:

English

Subjects (All):

Gene expression.

Gene therapy.

Physical Description:

1 online resource (296 pages)

Edition:

First edition.

Place of Publication:

New York : Nova Science Publishers, Inc., [2007]

Summary:

Esophageal cancer is malignancy of the esophagus. Esophageal tumors usually lead to dysphagia (difficulty swallowing), pain and other symptoms, and are diagnosed with biopsy. Small and localized tumors are treated with surgery, and advanced tumors are treated with chemotherapy, radiotherapy or combinations. This book presents an in depth look into the identification and treatment of esophageal cancer. This includes identification of esophageal squamous cells, epidemiology of esophageal cancer, atypical regenerative hyperplasia of the esophagus, genomic instability and gene therapy.

Contents:

Intro

NEW RESEARCH ONESOPHAGEAL CANCERHORIZONS IN CANCER RESEARCHVOLUME 40

CONTENTS

PREFACE

Chapter 1THE IDENTIFICATION, REGULATION ANDFUNCTIONAL STUDY OF THE DIFFERENTIALLYEXPRESSED GENES IN HUMAN ESOPHAGEALSQUAMOUS CELL CARCINOMA

ABSTRACT

INTRODUCTION

1. GENETIC OR ENVIRONMENTAL FACTORS, WHICH ISMORE IMPORTANT IN ESCC ETIOLOGY?

2. DISCOVERY OF CA2+-RELEVANT AND DIFFERENTIATIONASSOCIATED GENES DOWN-REGULATED IN ESOPHAGEALSQUAMOUS CELL CARCINOMA USING CDNA MICROARRAY

Materials and Methods

Patients and Samples

cDNA Library Construction

cDNA Array Construction

Hybridization and Image Processing

Sequencing Analysis of the Differentially Expressed Genes

Northern Blot and RT-PCR

Cell Culture and Transient Transfection

Results

Overview of Library Quality and EST Expression Pattern in ESCC

cDNA Microarray Analysis

Hierarchical Clustering Analysis

Alterations of Gene Expression in ESCC

Expression of GKLF in KYSE70 Stimulates SPRRs and KRT4 Transcription

Discussion

3. S100 GENE FAMILY IN HUMANESOPHAGEAL SQUAMOUS CELL CARCINOMA

Primer Design

Semi-Quantitative PCR and Image Analysis

Statistical Analysis

Immunohistochemistry

Semi-Quantitative RT-PCR

Correlation Analysis of 16 S100 Genes' Deregulation

Frequent Loss of S100A8 and S100A9 mRNA and Protein Expression in ESCCTissues

S100A8 and S100A9 Expression in Normal Esophageal Epithelium,Premalignant Lesions and Carcinomas

Intracellular Localization of S100A8 and S100A9 in Esophageal Tissues andCancer Cell Lines

S100A8 and S100A9 Expression Correlates with the Cell Differentiation

Discussion.

4. TRANSCRIPTION FACTOR C-JUN/AP-1 REGULATES THEEXPRESSION OF DIFFERENTIATION-ASSOCIATED GENES VIAMAPK PATHWAY

Introduction

Cell Culture and Materials

Preparation of Protein Extracts and Immunoblotting

Sample Collecting and Immunohistochemistry

Plasmid Constructions and Luciferase Assays

Electrophoretic Mobility Shift Assays (EMSA)

Northern Blot

Chromatin Immunoprecipitation (ChIP)

JNK Kinase Assay

The Endogenous Expression and Phosphorylation Levels of the Componentsof AP-1 Proteins

c-Jun/AP-1 Regulates Transcription and Expression of the Differentiation-Associated Genes

c-Jun/AP-1 DNA Binding Activity and Transactivation Activity are Mediated byTPA

TPA-Triggered Activation of c-Jun/AP-1 via MAPK Pathways

c-Jun/AP-1 Transactivates the Differentiation-Associated Genes byEnhancing the Binding of c-Jun to Promoters of the Genes

5. ALLELIC IMBALANCE OF CHROMOSOME 1Q IN ESCC: ANOVEL REGION OF ALLELIC LOSS AND SIGNIFICANTASSOCIATION WITH DIFFERENTIATION

DNA Extraction

Microsatellite DNA Markers of Chromosome 1q

PCR

Analysis of Allelic Imbalance

Statistic Analysis

6. FUNCTIONAL STUDY OF THE DIFFERENTIALLYEXPRESSED GENES IN ESCC

6.I. Overexpression of Stefin A in Human ESCC Cells Inhibits Tumor CellGrowth, Angiogenesis, Invasion and Metastasis

Cell Culture and Reagent

Preparation of Constructs and Transfection

Western Blot Analysis

Growth Kinetics Assay

Cathepsin B Activity Assay

Motility and Invasion Assay

Experiments in Nude mice

Factor VIII Staining and Microvessel Quantitation

High Invasive EC9706 Cells Show Low Stefin A Protein Level.

Transfection with Sense-stefin A Construct Inhibits Cell Growth in vitro

Stefin A-Transfected ESCC Cells Show Reduced Cathepsin B Activity and ReducedInvasiveness in vitro

Stefin A-transfected Cells Show Reduced Tumor Growth in vivo

Stefin A-transfected EC9706 Cells Show a significant Inhibition of Lung Metastasis in NudeMice

Stefin A-transfection Inhibits Tumor Angiogenesis

6.II. Stomatin-Like Protein 2 is Overexpressed in ESCC and Involved inRegulating Cell Growth and Cell Adhesion in Human ESCC

Tissue Specimens

Cell Culture

Semi-quantitative RT-PCR

Antibody Production and Western Blot Analysis

Immunohistochemical Staining

MTT Assay

Clonogenecity Assay

Flow Cytometry Assay

Experiments in Nude Mice

Cell Attachment Determination

Confocal Imaging

Overexpression of SLP-2 in ESCC

Overexpression of SLP-2 in Premalignant Lesions of ESCC Development

Screening of ESCC Cell Lines and Positive Transfectants

Transfectants with Antisense SLP-2 Inhibits Cell Growth in Vitro and Tumor Growth in Vivo

Transfectants with Antisense SLP-2 Inhibits Cell Attachment

CONCLUSION

ACKNOWLEDGEMENTS

REFERENCES

Chapter 2HUMAN PAPILLOMAVIRUS AND ESOPHAGEALSQUAMOUS CELL CARCINOMA

1. PREVALENCE OF HUMAN PAPILLOMAVIRUS IN HUMANESOPHAGEAL SQUAMOUS CELL CARCINOMA

1.1. Molecular Epidemiology of Human Papillomavirus in EsophagealSquamous Cell Carcinoma

1.1.1. Detection of HPV in ESCC Samples by PCR

1.1.2. Detection of HPV in Cancerous, Paracancerous and Normal MucosalTissues by PCR

1.1.3. Detection of HPV in the Histology of ESCC by ISH

1.2. Histology Suggesting HPV Infection

1.2.1. Papilloma and Papillomatous Lesions.

1.2.2. Koilocytotic Cells

1.2.3. Balloon Cells and Clear- Cell Acanthosis

1.3. Analysis of the Infection entry Pathway of HPV

1.4. Interaction of HPV with Cellular Biological Behaviors

2. IMMORTALIZATION AND MALIGNANT TRANSFORMATIONOF THE ESOPHAGEAL EPITHELIAL CELLS INDUCED BY HUMANPAPILLOMAVIRUS IN THE MULTISTAGE PROCESS

2.1. Establishment of SHEE Cell Line Induced by Human Papillomavirus

2.1.1. Construction of the Recombinant Plasmids

2.1.2. Cultivation of Human Embryonic Esophageal Mucosa

2.1.3. HPV18 E6E7-AAV Infection

2.2. Immortalization and Malignant Transformation of EsophagealEpithelial Cells in the Multistage process

2.2.1. The Preimmortal Stage [94]

2.2.2. The Immortalized Stage [95,96]

2.2.3. The Biphase Differential Stage [97,98]

2.2.4. The Malignantly Transformed Stage [99]

2.2.5. The Invasive and Metastasis Stage [100,101]

2.3. Biological Criteria used to Monitor the Developed Process ofImmortalization and Malignant Transformation in Cultured EsophagealEpithelial Cells

2.3.1. Cellular Proliferation, Differentiation and Cell Cycle

2.3.2. Telomere and Telomerase

2.3.3. p53, bcl-2, c-Myc and Ras Genes

2.3.4. Cytogenetic Abnormality

2.3.5. Tumorigenicity

2.3.6. cDNA Microarray Assay

2.4. The Promoters Effect on the Malignant Transformation in the InitialEsophageal Epithelial Cells

2.4.1. Human Papillomavirus in Synergy with TPA Induced the Formation ofMalignant Transformation of Esophageal Epithelial Cells [114]

2.4.2. Human Papillomavirus in Synergy with 60Cobablt Radiation PromotesMalignant Transformation of Esophageal Epithelial Cells [115]

2.4.3. The N-Nitrosopiperidine Promotes the Malignant Transformation ofEsophageal Epithelial Cells [116]

2.4.4. Sodium Butyrate Promotes the Malignant Transformation ofImmortalized Esophageal Epithelial Cells [117,118].

2.5. Conclusion

Chapter 3BIOLOGY OF ESOPHAGEAL CANCER:MECHANISMS OFRESISTANCE AND METASTASES

STAGING PROCEDURES

TREATMENT

MECHANISMS OF RESISTANCE

1- Resistance to DNA Damage Induction

Platinum Resistance

a- NER system

b- Mismatch repair

Fluoropyrimidine Resistance

2- Resistance to Apoptosis Induction

Extrinsic Pathway

Intrinsic Pathway

MECHANISMS OF METASTASIS

Tumor Circulating Cells (TCC)

Tumor Disseminated Cells (TDC)

Lymph Nodes

Bone Marrow

Lavage Citology of Cavities

Macroscopic Metastases

Stem Cells

ACKNOWLEDGEMENT

Chapter 4PRECLINICAL EVIDENCE FOR THE USE OFKEYHOLE LIMPET HEMOCYANIN INBARRETT'S ADENOCARCINOMA

METHODS

Cell Death Detection ELISA Plus

Proteomics

Two Dimensional Electrophoresis (2DE)

Matrix-Assisted Laser Desorption Ionization Time of Flight MassSpectrometry (MALDI-TOF-MS).

RESULTS

Decreased Cell Viability with KLH

Apoptosis Induced by KLH

Changes in Protein Expression with KLH

Chapter 5CHEMORADIATION TREATMENT IS EFFECTIVEFOR THE PALLIATION OF MALIGNANTDYSPHAGIA IN OESOPHAGEAL CANCER

NONRANDOMISED TRIALS OF EXTERNALBEAM RADIATION THERAPY

NONRANDOMISED TRIALS OF CHEMO RADIATION

PHASE III TRIALS OF RADIATIONVERSUS CHEMO RADIATION

Radiation Versus Concomitant Chemo Radiation

Radiation Versus Sequential Chemo Radiation

METHODS AND MATERIALS

Relief of Dysphagia

Re-Intervention

Toxicity

Survival

Quality of Life Assessments

DISCUSSION

1. How Effectively will Dysphagia be Relieved and how long will it take toRelieve?.

2. Does Chemotherapy add to the Benefit of Radiation in the Palliation ofMalignant Dysphagia?.

Notes:

Includes bibliographical references and index.

Description based on print version record.

Other Format:

Print version: Carminati, Antonio New Research on Esophageal Cancer

ISBN:

979-88-911-3340-2