Down-regulation of Forkhead Box M1 Transcription Factor Leads to the Inhibition of Invasion and Angiogenesis of Pancreatic Cancer Cells. Zhiwei Wang, Sanjeev Banerjee, Dejuan Kong, Yiwei Li, and Fazlul H. Sarkar Department of Pathology, Karmanos Cancer Institute, Wayne State University, Detroit, Michigan Cancer Res 2007; 67: (17). September 1, 2007

1. Zhiwei Wang, Sanjeev Banerjee, Dejuan Kong, Yiwei Li and Fazlul H. Sarkar Department of Pathology, Karmanos Cancer Institute, Wayne State University, Detroit, Michigan Design by ChauChanLao 2009.03 Cancer Res 2007; 67: (17). September 1, 2007 IF: 7.672

2. Introduction

6. www.cs.uiuc.edu

8. FOXM1 regulate transcription of many cell cycle genes (Cdc25A, Cdc25B, cyclin B, cyclin D1, p21, p27) http://mcb.asm.org

9. Angiogenesis ( 血管新生 ) Cell and molecular biology, Gerald Karp, p.685 http://ir.progenics.com

10. Vascular endothelial growth factor (VEGF) http://www.researchvegf.com FoxM1 regulates VEGF signaling in various cell types

11. Migration and Invasion Migration ( 遷移 ) Invasion ( 入侵 )

12. Cell and molecular biology, Gerald Karp, p.240, p.244 Type IV collagen network

15. The aim in this article Whether FoxM1 could be an effective therapeutic target for PC ?

17. Experimental Design MTT test Flow cytometry ELISA Angiogenesis assays Migration and Invasion assay

18. RNA interference (RNAi) wikipedia.org

19. Gene transfection FoxM1 siRNA, siRNA control or FoxM1 cDNA plasmid Lipofectamine 2000

20. PC Cells Incubate the cells

21. PC Cell DNA

22. To determine the baseline expression of FoxM1 in PC cell lines Fig.1

23. To examine the relative mRNA levels of FoxM1 in three PC cell lines Fig.1

24. http://biochemistry.yonsei.ac.kr/

26. To determine the efficacy of FoxM1 siRNA for knockdown FoxM1 mRNA and protein Fig.1 CS : control siRNA FS : FoxM1 siRNA

27. To examine the effect of FoxM1 siRNA on cell growth of PC cells Fig.2 wikipedia.org MTT test

28. To examine the effect of FoxM1 cDNA on cell growth of PC cells CP : control plasmid FP : FoxM1 cDNA plasmid Fig.1D Fig.2

30. To further investigate the growth-inhibitory effect of FoxM1 knockdown in PC cells Fig.3 Cell cycle anatysis

31. To examine the level of expression of several known G 0 -G 1 cell cycle regulatory factors Fig.4

33. To investigate whether MMP-9 and MMP-2 were down-regulated by FoxM1 siRNA in PC cell lines Fig.5

34. To examine whether the down-regulation of FoxM1 could lead to a decrease in MMP-2 and MMP-9 activity in PC cells Fig.5 ELISA

36. To investigate whether FoxM1 siRNA has any effect on reducing the level of uPAR (urokinase-type plasminogen activator receptor), which are able to regulate the MMP-9 activity in PC Fig.5 FoxM1 siRNA reduced uPAR gene transcription and translation

37. To test the effects of FoxM1 down-regulation on PC cell migration and invasion, using BD BioCoat Tumor Invasion Assay System www.bdbiosciences.com

38. Fig.6 Cell migration and invasion assay

39. Stain http://www.dojindo.cn wikipedia.org Calcein-AM Calcein 細胞內酯酶水解

41. To explore whether FoxM1 siRNA reduced VEGF activity Fig.5 FoxM1 siRNA decreased VEGF acivity (Overexpression FoxM1 increased VEGF acivity, data not shown)

42. To test whether conditioned media from FoxM1 siRNA-transfected cells could reduce the tube formation Fig.6 Matrigel in vitro HUVECs tube formation assay

44. Summary

45. Conclusion The down-regulation of FoxM1 could potentially be an effective therapeutic approach for the inactivation MMP-2, MMP-9, and VEGF, which is likely to result in the inhibition of cell growth, migration, invasion, angiogenesis, and metastasis of PC.

47. wikipedia.org