Title Page
ABSTRACT
Contents
INTRODUCTION 10
MATERIAL & METHODS 12
1. TCGA data analysis 12
2. Tissue microarray (TMA) and immunohistochemistry (IHC) 12
3. Surgical CCA mouse model 12
4. Reverse transcription-PCR, mutation analysis, and microarray 12
5. Cell culture and transfection 13
6. Immunoblotting 13
7. Trypan blue exclusion assay 14
8. Thiazolyl blue tetrazolium bromide (MTT) assay 14
9. Flow cytometric cell cycle analysis 14
10. Transwell assay 14
11. Wound-healing assay 15
12. Quantitative real-time PCR 15
13. Statistical analysis 15
RESULTS 16
1. PIF1 is highly expressed in CCA 16
2. PIF1 promotes the proliferation of CCA cell lines 16
3. PIF1 promotes the invasiveness of CCA cell lines 16
4. PIF1 regulates the TGF-β/SMAD4 signaling pathway in CCA cell lines 17
DISCUSSION 25
CONCLUSION 27
REFERENCES 28
국문 요약 33
Table 1. List of reverse transcription PCR primer sequences. 23
Table 2. List of quantitative real-time PCR primer sequences. 24
Figure 1. Upregulated expression of PIF1 in human and mouse CCA. 18
Figure 2. Knockdown of PIF1 inhibits the proliferation of CCA cell lines. 19
Figure 3. Knockdown of PIF1 reduces the invasiveness of CCA cell lines. 20
Figure 4. Inhibition of the TGF-β/SMAD4 signaling pathway by PIF1 knockdown in CCA cell lines. 21
Supplementary Figure. (A) Mutation status of PIF1 in CCA cell lines. (B) Cell cycle analysis in HUCCT1 and SNU-478 cell lines. (C) Western blot analysis of PIF1 and SMAD4 in HUCCT1 cell line.... 22