1. Ρύθμιση κυτταρικού κύκλου & Εισαγωγή στην κυτταρική σηματοδότηση Γιώργος Ρασιδάκης Επ. Καθηγητής Παθολογικής Ανατομικής, Ιατρική Σχολή ΕΚΠΑ Adj. Assistant Professor, M.D. Anderson Cancer Center (USA)

3. Interphase (12-24h, RNA and protein synthesis, cell growth) Gap 0 (G0) cell quit dividing temporarily or more permanently Gap 1 (G1) RNA and protein synthesis; G1 checkpoint S (synthesis) DNA replication Gap 2 (G2) protein synthesis and cell growth; G2 checkpoint Mitosis or M Phase (1-2h) division into two similar daughter cells; metaphase checkpoint Cell Cycle and Division

4. G1 G2 S Cell grows Cell division M DNA replication Cell prepares to divide C C C Metaphase checkpoint G1 checkpoint G2 checkpoint G0 Cells are responsive To mitogenic signals R

5. G2 S Cell grows Cell division M DNA replication Cell prepares to divide C C C Cyclin D1/2/3 Cdk4/6 RbE2F InactiveInactive P Rb P E2F Active S Phase Genes E2F DP Cyclin E Cdk2 Cyclin A Cdc2 Cyclin A Cdc2 Cyclin B INK4 p15,p16 p18,p19 INK4 p15,p16 p18,p19 CIP/KIP p21, p27 p57 CIP/KIP p21, p27 p57 G1

9. Hypermethylation of INK4 Genes in Cancer

12. Lymphomas “Low grade”“High grade” proliferation apoptosis (e.g. CLL/SLL, FL gr.1)(e.g. DLBCL, BL)

13. Burkitt Lymphoma

14. c-MYC promotes G1–S progression through gene activation and repression Gene activation Gene repression p15 p21 p27

17. Retinoblastoma

18. Inactivation of retinoblastoma protein

19. Loss of heterozygosity of Rb locus (13q14)

22. Advantages of ubiquitin-regulated proteolysis

24. Important inhibitor of cell cycle progression Haplo-insufficient tumor suppressor gene Rarely mutated or silenced in cancers p27 expression is inversely related to proliferation in many tumor types Low p27 levels due to protein degradation are associated with tumor aggressiveness and poor prognosis in cancer p27 Kip1

25. p27 Kip1 Regulation Post-translational regulation Phosphorylation at T187 / T157 / S10 Degradation through the ubiquitin / proteasome pathway (Cks1/Skp2) Cul1 Cyclin E Cdk2 p27 Cyclin E Cdk2 P Cyclin E Cdk2 P Cyclin E Cdk2 Ub Ub Ub Skp2 Ubc3 Skp1 Cks1 p27 degradation

26. p27 Kip1 Cdk-bindingsite P-richCdk-bindingsiteJAB1-bindingsite12788 151 169 198 CN NLS Cyclin-bindingsite(25-51)Cdk-bindingsite(60-79) 97 S10 KISkinaseInteractingstathmin T157 Impairsnuclearimport PKB/Akt Phosphorylationsites: T187 Cdk2 DegradationUb./Proteasomepathway Stability In G1

27. 198 JAB1-bindingsite p27 Kip1 Cdk-bindingsite P-richCdk-bindingsite 12788 CN Cyclin-bindingsite(25-51)Cdk-bindingsite(60-79) 97 S10 KISkinaseInteractingstathmin Phosphorylationsites: T187 Cdk2 DegradationUb./Proteasomepathway Stability In G1 169 T157 Impairsnuclearimport PKB/Akt 151NLS

28. Cyclin D Cdk4/6 Cyclin E Cdk2 CytoplasmNucleus Akt DNA Cyclin E Cdk2 E2F Cell cycle progression STOP Akt activating kinases Akt Cyclin A Cdk 2 Akt Activation Induces Cell Cycle Progression Through p27 Kip1 Downregulation

29. Figure 8.16a The Biology of Cancer (© Garland Science 2007)

30. Figure 8.16b The Biology of Cancer (© Garland Science 2007)

31. Σηματοδοτική οδός JAK / STAT

32. James E. Darnell, Jr., The Rockefeller University

34. Σηματοδοτική οδός PI3K / AKT / mTOR

35. Corradetti and Guan, Oncogene, 2006 Negative feedback loop mTOR-Rictor Activates AKT/PKB

36. Hay and Sonenberg, Genes Dev. 2004 mTOR Controls Translation Through Phosphorylation of p70S6K and 4E-BP1 mTOR

37. Hay and Sonenberg, Genes Dev. 2004 4E-BP1 Ribosome recruitment to mRNA (5’ end) eIF-4E binds the 5’ cap structure of mRNA 4E-BP1 PPPP

38. Σηματοδοτική οδός RAS / MEK / ERK

39. Figure 6.12a The Biology of Cancer (© Garland Science 2007)

40. Figure 6.14 The Biology of Cancer (© Garland Science 2007)

41. Figure 6.15 The Biology of Cancer (© Garland Science 2007)

42. AKT PI3K MEKMDM2 p53 RAS ERK mTOR PTEN survival GF- R RAF GF Proliferation BAD

43. AKT PI3K MEKMDM2 p53 RAS ERK mTOR PTEN Επιβίωση GF- R NF RAF GF Πολλαπλασιασμός Μεταλλάξεις RAS 20-50% Μεταλλάξεις PIK3CA 20% BAD Μεταλλάξεις BRAF Μεταλλάξεις p53 50%

44. PTEN TSC1/TSC2 Rheb AKT PI3K PDK1 PIP3 p70S6K4E-BP1 eIF4ErpS6 GβL mTOR RAPTOR GβL mTOR RICTOR Protein translation Flt3 GRB2 RAS ERK1/2 RAF1 MEK1/2 SHC SHIP SHP GAB2 GRB2 Cyclins (e.g. D1-3, A) CDK Inhibitors (e.g. p21, p27) JAK SHP1 SOCS SHP2 STAT3 Oncogenic kinases STAT3 Transcription

45. Σηματοδοτική οδός NFkB

46. Figure 6.29a The Biology of Cancer (© Garland Science 2007)

47. Thank you