Title page
Acknowledgement
Contents
Abstract 9
1. INTRODUCTION 11
2. MATERIALS AND METHODS 13
2.1 RNA Isolation 13
2.2 cDNA Synthesis 14
2.3 Suppression Subtractive Hybridization (SSH) 14
2.4 Cloning and Analysis of the subtracted cDNA 15
2.5 Reverse transcription-polymerase chain reaction (RT-PCR) 16
2.6 Patients and tissue specimens 17
2.7 Serum Stimulation 17
3. RESULTS 18
3.1 Identification of genes overexpressed in human immortalized cell using SSH 18
3.2 Verification of 58 gene in immortalized cell, cancer cell lines and some lung cancer tissues by RT-PCR 20
3.3 Serum stimulation 23
4. DISCUSSION 27
5. CONCLUSION 30
REFERENCES 31
ABSTRACT IN KOREAN 36
Table 1. Identification of known tumor specific genes from SSH libraries. 21
Fig. 1. Reduction of G3PDH abundance by PCR-select subtraction. 19
Fig. 2. Sequence of Clone 58 22
Fig. 3. Expression of clone 58 in WI-38, WI-38VA13 (a) and various cell lines (b). 24
Fig. 4. Clone 58 expression in non-small cell lung carcinoma tissues. 25
Fig. 5. Serum stimulation. 26