Title Page

Contents

국문초록 7

ABSTRACT 9

Introduction 11

Materials and Methods 14

Results 19

Effects of 2-D08 on cell viability of Ut-LMS cells 19

2-D08 inhibits colony formation of Ut-LMS cells 19

2-D08 suppresses the proliferation of Ut-LMS cells in vitro 20

Effect of 2-D08 on the protein expression related to cell proliferation and apoptosis in Ut-LMS cells 20

2-D08 treatment inhibits the degradation of p21 by Ub-proteasome pathway in Ut-LMS cells 21

The impact of 2-D08 on the expression of differentiation marker proteins in Ut-LMS cells 22

Discussion 23

References 39

Figure 1. Chemical structure of 2-D08. 27

Figure 2. 2-D08 decreased cell viability in the SK-LMS-1 cells. (A, B) The cells were treated with 2-D08 in various concentrations (0–100 μM) for 24 or 48 h. After the end of... 28

Figure 3. 2-D08 decreased colony formation ability in the SK-LMS-1 cells. (A) Cells underwent a 7 d treatment with 2-D08 at concentrations ranging from 0 to 100 μM).... 29

Figure 4. Inhibition of SK-LMS-1 cell proliferation by 2-D08. (A) During a 48 h period, SK-LMS-1 cells underwent treatment with 20 and 50 μM concentrations of 2-D08, and... 30

Figure 5. The cell proliferation assay using BrdU labeling. BrdU assay was conducted following 24 and 48 h treatment of SK-LMS-1 with 2-D08 (20 or 50 μM). Cells were... 31

Figure 6. The effects of 2-D08 on the expression of cell proliferation genes in SK-LMS- 1 cells. (A) Expression of p21, p27, and p53 protein in SK-LMS-1 cells exposed to 2-... 32

Figure 7. The effects of 2-D08 on the expression of apoptosis-related genes in SK-LMS- 1 cells. (A) Effect of 2-D08 on the Caspase-3 activity in the SK-LMS-1 cells. (B) Western... 33

Figure 8. Proteasome inhibition increased the steady-state levels of p21 in SK-LMS-1 cells. (A) Cells were exposed to the indicated concentrations of MG132 for 4 h, and p21,... 34

Figure 9. 2-D08 inhibits p21 ubiquitination in SK-LMS cells. (A) Cells were transfected with Ub expression vector and were treated with 50 μM of 2-D08 for 48 h. Cell lysates... 35

Figure 10. The analysis of contractile SMC-specific proteins expression was conducted using western blot analysis. (A) The expressions of α-SM-actin, TAGLN, and Calponin... 36

Figure 11. Expression of contractile SMC-specific proteins analyzed using quantitative real-time PCR analysis. The mRNA levels of α-SM-actin, TAGLN, and Calponin 1 were... 37

Figure 12. Diagram. 2-D08 inhibits cell proliferation by regulating the stabilization of p21. 38