ABSTRACT
PARTI Identification of the Functional Role of Oncogene AFIQ in Breast Cancer Metastasis Based on Cdan Microarray Analysis
INTRODUCTION
MATERIALS and METHODS
Cell Lines and Animals
Bacterial strain
RT-PCR Analysis
Construct of Human AF1Q Expression Vector
Preparation of Competent E. coli Cells for Transformation
Transformations
Identification of Recombinant Plasmid in Bacterial Colonies by PCR
Preparation of Plasmid DNA
Restriction Enzyme Digestion of Recombinant Plasmid DNA
Sequencing Analysis
Stable Transfections
Expression Analysis of AF 1Q Transfectants
In Vitro Invasion Assays
RT-PCR analysis
Western Blot Analysis
Zymographic analysis
Measurement of MMP-2 and MMP-9 by ELISA
RNA interference (RNAi) Experiments
Tumorigenicity and Metastasis Assays in Nude Mice
Western Blot Analysis of MMP-2, MMP-9, Est-l, and Rhoc Expression
Expression of AF1Q genes in Clinical Breast Cancer Specimens
Statistical Analysis
RESULTS
Validation of the results of cDNA microarray analysis
Construct of Human AF 1Q Expression Vector
Stable transfection and expression analysis
Overexpression ofAF 1Q Enhances the Invasive Potential of MDA-MB-435 Cells In Vitro through Regulation of MMP-2,MMP-9,Est-1,and Rhoc Expression
Reduced AF 1Q Expression Decreases Tumor Invasion In Vito and Down-regulates MMP-2,MMP-9,Ets-1,and Rhoc Protein Expression
OverexpressionofAF1QEnhancesTumorGrowthandPulmonaryMetastasis Athymic Mice
Expression of AF1Q Gene in Clinical Breast Cancer Specimens
DISCUSSION
REFERENCES
Tables and Figures
PART ⅡIdentification of Breast Caner Metastasis-AssociaPioteins by Using Two-Dimensional Gel Electrophoresis andHigh Performance Liquid Chromatography-Ion Trap-Mass Spectrometry
INTRODUCTION
MATERIALS and METHODS
Chemicals
Cell culture and sample preparation for 2-DE
2-DE and Silver-staining
Image Acquisition and Statistical Analysis
In-gel Digestion
LC-IT-MS
Protein Identification and Database Searching
RT-PCR Analysis
Western Blotting Analysis
RESULTS
Proteome Differential Expression between MDA-MB-435HM and its parental
Identification of Differentially Expressed Proteins by MS/MS
Verification of Differentially Expression Proteins by RT-PCR and Western
DISCUSSION
REFERENCES
Tables and Figures
综述 蛋白质级学技术在乳腺癌研究中的应用
博士后期间发表论文、参加学术会议、所获奖励与基金
致谢
