Title Page
Contents
ABSTRACT 10
INTRODUCTION 12
MATERIALS AND METHODS 15
1. Cell culture and reagent 15
2. Zebrafish maintenance and husbandry 15
3. Microinjection of cancer cells in zebrafish 16
4. Drug treatment 16
5. Imaging and microscopy 16
6. Cell viability assay 17
7. Acridine orange staining and microscopy 17
8. RNA extraction and reverse transcript (RT)-PCR 18
9. Protein extraction and Western blotting 18
10. Whole-mount immunostaining and microscopy 19
11. Statistical analysis 20
RESULTS 22
1. The sensitivity to gefitinib in normal human bronchial epithelial cells and NSCLCs 22
2. Validation of anti-cancer effect of gefitinib using zebrafish xenograft models 25
3. Visualization of NSCLC cell growth and dynamics in zebrafish xenografts 29
4. Gefitinib regulates EGFR/AKT signaling pathway in NSCLC xenografted zebrafish embryos 32
5. RSL3 induces anti-cancer activity through GPX4 inhibition in GR-NSCLC cells 35
6. The effects of A1 are mediated through activation of p53 on GR-NSCLC cells 42
7. Heterogeneous immunophenotypes of different lung cancers in xenograft tumors 49
DISCUSSION 53
REFERENCES 56
ABSTRACT IN KOREAN 61
Table 1. List of primer sequences used for RT-PCR analysis 21
Table 2. Gene status of normal human bronchial epithelial cell and NSCLC cell lines 23
Figure 1. Gefitinib induces apoptosis in GS-HCC827 cells. 24
Figure 2. Gefitinib inhibits the growth of xenografted HCC827 cells in... 28
Figure 3. The growth and proliferation of HCC827 cells were... 31
Figure 4. Gefitinib reduced phosphorylation of EGFR, AKT, and MEK... 34
Figure 5. RSL3 acts as an anti-cancer drug in GR-H23 and -H1299 xenografts. 40
Figure 6. A1 acts as an anti-cancer drug in GR-A549 and -H460 xenografts. 47
Figure 7. Heterogeneous immunostaining of mixed NSCLC xenografts. 52