Title page
Acknowledgement
Contents
Abbreviation 10
Abstract 11
1. INTRODUCTION 12
2. MATERIALS AND METHODS 14
2.1 Tissue Materials 14
2.2 RNA extraction and SMART PCR cDNA Synthesis 14
2.3 Suppression Subtraction Hybridization (SSH) 15
2.4 Cloning and Analysis of the subtracted cDNA 19
2.5 Reverse transcription-polymerase chain reaction (RT-PCR) 19
2.6 Serum Stimulation 20
3. RESULTS 21
3.1 Identification of genes overexpressed in glioblastoma using SSH 21
3.2 Verification of 7clones in brain tumor tissues and cancer cell lines by RT-PCR 21
3.3 Serum Stimulation in Normal WI-38 Fibroblasts 25
4. DISCUSSION 30
5. CONCLUSION 33
REFERENCES 34
ABSTRACT IN KOREAN 39
Table 1. The primer used in RT-PCR analysis 22
Table 2. Identification of known tumor specific genes from SSH libraries 23
Table 3. Identification of unknown tumor specific genes from SSH libraries 24
Figure 1. Scheme of the SSH method 16
Figure 2. Expression of brain 7 clones mRNA from human glioblastoma tissues by RT-PCR 26
Figure 3. Expression of7 clones mRNA from human cellline by RT-PCR 27
Figure 4. WI-38 fibroblast cells time course of 7 clones 28