| ACKNOWLEDGEMENT | 第7-8页 |
| 摘要 | 第8-9页 |
| ABSTRACT | 第9页 |
| REVIEW Non-Small Cell Lung Cancer and TGF-Beta/Smad Signaling Pathway | 第11-36页 |
| Chapter 1 Non-Small Cell Lung Cancer and Tumor Suppressor Genes | 第12-19页 |
| 1.1 Introduction to non-small cell lung cancer | 第12-14页 |
| 1.1.1 Lung cancer epidemiology | 第12-13页 |
| 1.1.2 Staging of non-small cell lung cancer | 第13-14页 |
| 1.2 Molecular approaches to lung cancer evaluation | 第14-16页 |
| 1.3 Identification of tumor suppressor genes | 第16-19页 |
| Chapter 2 TGF-Beta/Smad Signaling Transduction Pathway and Tumorigenesis | 第19-30页 |
| 2.1 TGF-beta superfamily members and TGF-beta receptors | 第19-23页 |
| 2.1.1 Transforming growth factor-beta superfamily members | 第19页 |
| 2.1.2 Biological function of transforming growth factor-beta | 第19-20页 |
| 2.1.3 Transforming growth factor-beta receptors | 第20-22页 |
| 2.1.4 Genomic organization of TGFBRII and TGFBRI genes | 第22-23页 |
| 2.2 Smad family members and Smad2 genomic structure | 第23-24页 |
| 2.2.1 Introduction to Smad family members | 第23-24页 |
| 2.2.2 Genomic structure of Smad2 gene | 第24页 |
| 2.3 TGF-beta/Smad signaling pathway in epithelial cells | 第24-25页 |
| 2.4 TGF-beta/Smad signaling pathway and cancers | 第25-30页 |
| 2.4.1 Mutations of TGFBRII and TGFBRI in cancers | 第26-28页 |
| 2.4.2 Mutations of Smads in cancers | 第28-30页 |
| References | 第30-36页 |
| PART Ⅰ Mutational Analysis of the Key Effectors of TGF-Beta/Smad Signaling Pathway in Primary Non-Small Cell Lung Cancer | 第36-76页 |
| Chapter 1 Introduction | 第37-41页 |
| 1.1 Microsatellite instability and non-small cell lung cancer | 第37页 |
| 1.2 Mutations of TGFBRII, TGFBRI and Smad2 genes in NSCLC | 第37-39页 |
| 1.3 Design of this part of research work | 第39-41页 |
| Chapter 2 Materials and Methods | 第41-62页 |
| 2.1 Materials | 第41-49页 |
| 2.1.1 Specimens | 第41-43页 |
| 2.1.2 Markers and primers for MSI analysis | 第43页 |
| 2.1.3 Reagents and solutions | 第43-48页 |
| 2.1.4 Equipments and tools | 第48页 |
| 2.1.5 Internet resources and softwares | 第48-49页 |
| 2.2 Methods | 第49-62页 |
| 2.2.1 Isolation of genomic DNA | 第49-52页 |
| 2.2.1.1 Isolation of genomic DNA from tissue | 第49-50页 |
| 2.2.1.2 Extraction of genomic DNA from blood | 第50-52页 |
| 2.2.2 Amplification of DNA | 第52-54页 |
| 2.2.3 Genotyping for MSI by GeneScan | 第54-55页 |
| 2.2.4 Mutational screen by DGGE and SSCP | 第55-56页 |
| 2.2.5 Confirm mutations and SNPs by sequencing | 第56-60页 |
| 2.2.5.1 Direct DNA sequencing | 第56-58页 |
| 2.2.5.2 Cloning sequence | 第58-60页 |
| 2.2.6 Geuotyping SNPs by RFLP | 第60-61页 |
| 2.2.7 Statistical analysis | 第61-62页 |
| Chapter 3 Results | 第62-68页 |
| 3.1 MSI analysis for genomic DNA | 第62-63页 |
| 3.2 Mutational analysis of TGFBRII, TGFBRI AND Smad2 genes | 第63-68页 |
| Chapter 4 Discussion and Conclusions | 第68-72页 |
| 4.1 MSI and mutation analyses for TGFBRII gene | 第68-69页 |
| 4.2 Analyses of mutation and polymorphism within TGFBRI gene | 第69-70页 |
| 4.3 Mutational analysis of Smad2 gene | 第70-71页 |
| 4.4 Conclusions | 第71-72页 |
| References | 第72-76页 |
| PART Ⅱ Defected Expression of TGFBRII Associated with CpG Methylation of TGFBRII Gene Promoter in Primary Non-Small Cell Lung Cancer | 第76-98页 |
| Chapter 1 Introduction | 第77-79页 |
| Chapter 2 Materials and Methods | 第79-85页 |
| 2.1 Materials | 第79-80页 |
| 2.1.1 Specimens | 第79页 |
| 2.1.2 Reagents and solutions | 第79-80页 |
| 2.1.3 Equipments and tools | 第80页 |
| 2.2 Methods | 第80-85页 |
| 2.2.1 Isolation of genomic DNA from tissue | 第80页 |
| 2.2.2 Immunohistochemistry analyses of TGFBRII and TGFBRI | 第80-81页 |
| 2.2.3 Immunohistochemistry analysis of Smad2 | 第81页 |
| 2.2.4 Evaluation of the immunohistochemical staining | 第81页 |
| 2.2.5 DNA modification before methylation-specific PCR (MSP) analysis | 第81-82页 |
| 2.2.6 MSP analysis for TGFBRII and TGFBRI | 第82-83页 |
| 2.2.6.1 MSP analysis for TGFBRII | 第82页 |
| 2.2.6.2 MSP analysis for TGFBRI | 第82-83页 |
| 2.2.7 RNA extraction and RT-PCR analysis | 第83-85页 |
| Chapter 3 Results | 第85-92页 |
| 3.1 Immunohistochemistry for TGFBRII | 第85-86页 |
| 3.2 Immunohistochemical staining for TGFBRI | 第86-88页 |
| 3.3 Immunohistochemical staining for Smad2 | 第88页 |
| 3.4 MSP analysis for TGFBRII and TGFBRI genes | 第88-90页 |
| 3.5 RT-PCR analysis For TGFBRII | 第90-92页 |
| Chapter 4 Discussion and Conclusions | 第92-95页 |
| 4.1 Defected expression of TGFBRII associated with CpG methylation of TGFBRII gene promoter in NSCLC | 第92-93页 |
| 4.2 Defected expression of TGFBRI in NSCLC | 第93-94页 |
| 4.3 Conclusions | 第94-95页 |
| References | 第95-98页 |
| Publications | 第98-99页 |
