Study on the Influence of Genetic Polymorphisms on Gastric Cancer Susceptibility in Northern Jiangsu(China)

声明

ABSTRACT

Contents

Chapter 1 Introduction

1．1．Gastric cancer epidemiology

1．2．Genetic variations and gastric cancer risk

1．3．Reactive oxygen species(ROS)

1．3．1．Biochemistry of reactive oxygen species(ROS)

1．3．2．Role of ROS in the development of human cancer

1．3．3．Reactive oxygen species(ROS)and DNA repair

1．4．Base Excision Repair(BER)

1．4．1．Basic mechanisms for base excision repair(BEll)

1．4．2．Role of DNA glycosylases in BER

1．4．3．BER in Mitochondria

1．5．Genotyping of single nucleotide polymorphisms(SNPs)

1．5．1．Hybridization based methods

1．5．2．Enzyme based methods

1．5．3．Allele-specific ligation

1．5．4．Single strand conformation polymorphism(SSCP)

1．5．5．Temperature gradient gel electrophoresis(TGGE)

1．5．6．Denaturing high performance liquid chromatography(DHPLC)

1．6．Single Nucleotide Polymorphisms(SNPs)

1．6．1．Apurinic／apyrimidinic endonuclease 1(APE 1)

1．6．2．Nei-like 2(NEIL2)

1．6．3．Mouse Double Minute 2 gene(MDM2)

1．6．4．CYP2E1

Chapter 1 summary

REFERENCES

Chapter 2 Materials and Instruments

2．1．Materials

2．1．1．Chemicals，reagents，primers，and probes

2．1．2．DNA Extraction

2．1．3．Total RNA extraction

2．2．Preparation and Characterization of Magnetic Nanomaterials

2．2．1．Preparation and characterization of Fe3O4 magnetic nanoparticles(MNPs)

2．2．2．Preparation and characterization of SiO2@Fe3O4 core-shell MNPs

2．2．3．Preparation and characterization of three-layer core-shellFe3O4@SiO2@Au MNPs

2．3．Instruments

Chapter 2 summary

REFERENCES

Chapter 3 Genetic Polymorphisms and Gastric Cancer Risk in Northern Jiangsu(China)Based on Magnetic Nanomaterials Genotyping

3．1．NEIL2(rs804270)SNP genotyping using single base extension(SBE)assay based on core-shell Fe3O4@SiO2@Au MNPs

3．2．FunctionaUzation and immobilization of Fe3O4@SiO2@AuMNPs with streptavidin

3．3．NEIL2 PCR and SBE genotyping reactions

Chapter 3 summary

REFERENCES

Chapter 4 Genetic Polymorphisms and Gastric Cancer Risk in Northern Jiangsu(China)Based on RT-PCR Genotyping Technique

4．1．TaqMan RT-PCR SNP genotyping assay

4．2．Statistical analysis

4．3．Frequency distribution of genotypes among gastric cancer cases and controls

4．4．Association of genotypes and gastric cancer risk

4．5．NEIL 2(rs804270)SNP and its association with susceptibility to gastric cancer

4．6．APE1(rs2275008)and its association with susceptibility to gastric cancer

4．7．CYP2E1 SNPs and their association with susceptibility to gastric cancer

4．8．MDM2 SNP(rs2279744)and its association with susceptibility to gastric cancer

4．9．SYBR Green Ⅰ?Quantitative Real-time PCR(qPCR)

4．10．Differential expression of NEIL2 mRNA among NEIL2 genotypes

Chapter 4 summary

REFERENCES

Chapter 5 Conclusion and Prospective

Appendix

ACKNOWLEDGEMENTS