Cancer, Cell communication and the Cell Cycle I . Cell Communication – Chapter 16

1. Cancer, Cell communication and the Cell Cycle I. Cell Communication – Chapter 16 The major signaling pathways relevant to cancer You will not be responsible for: Specific downstream signaling pathways Questions in this chapter you should be able to answer: Chapter 16:1 - 10 11all but e, 12,13,16,17,18, 19, 20, 22, 24, 25 Cancer and Cell Communication

2. The life and death of cells Why do cells eventually die? -- infection -- genetic mutation -- potentially harmful -- reach replicative limit How do cells die? Apoptosis vs Necrosis Apoptosis Cancer and Cell Communication

3. Cancerous cells have two key properties • Capacity for perpetual cell division - Cells replicate through a process called the Cell Cycle - Cell replication is carefully regulated 2) Loss of ability to undergo apoptosis - mutations accumulate over time Some “oncology” terminology Benign tumor Malignant tumor Metastasis Primary vs secondary tumor Cancer Cancer and Cell Communication

4. Metastasis requires many cellular changes and is also an evolutionary process Metastasis Cancer and Cell Communication

5. We will focus on mutations that alter 3 key cell properties and “set the stage” for metastasis  Mutations that trigger cell proliferation  activation of “Oncogenes”  Mutations that disable DNA error detection & apoptosis  disabling of “Tumor suppressor proteins”  Mutations that confer immortality  activation of “Telomerase” Cancer and Cell Communication

6. How do cells communicate with each other? Signaling mechanisms Signaling responses Cancer and Cell Communication

7. EGF What types of molecules carry signals to cells? 1) Gases (really small) NO, H2S, CO 2) ‘Smallish’ organic molecules steroids neurotransmitters [drugs/poisons (nicotine, phytohormones, etc)] 3) Peptide hormones (much bigger) Cancer and Cell Communication

8. Where are the receptors? Intracellular receptors vs Cell-surface receptors Cancer and Cell Communication

9. Oncogenes are mutated genes for proteins that control when cells divide For example… Cell membrane receptors for hormones that signal cells to divide Epidermal Growth Factor (EGF) binds to HER-receptor When bound to EGF, the part of receptor inside membrane signals cell to divide…. How? Cancer and Cell Communication

10. Activated receptors cause activation of other proteins inside the cell Convey a signal to nucleus Activate gene expression A cellular response -- e.g. cell division Many of these are potential oncogenes Receptor Signaling Cancer and Cell Communication

11. Oncogenes code for mutated receptors or signaling proteins that are trigger cell division inappropriately Normal genes code for proteins that function normally Mutated genes can code for abnormal proteins If “always on” = an oncogene Cancer and Cell Communication

12. How do cell surface receptors ‘signal’? Signal transduction Pathways Signaling proteins Secondary Signals -- cAMP, Ca++, DAG, IP3 FSH & Receptor Cancer and Cell Communication

13. Signaling pathways can interact Multiple signals Processed simultaneously Activating or inhibiting Signal integration Cancer and Cell Communication

14. What are the three types of cell surface receptors? = Ligand-gated channel Cancer and Cell Communication

15. How are G-proteins activated? “7-pass” receptors -- Hundreds of different types -- triggering enumerable different cytoplasmic processes Examples Glucagon – activates glucose release by liver Lutenizing Hormone (LH) – triggers progesterone release from ovary Adrenalin (epinephrine) – increases heart rate Allergen – mast cell degranulation G-protein-linked receptors Cancer and Cell Communication

16. Acetylcholine acts at a G-protein-linked receptor on heart muscle to make the heart beat more slowly by the effect of the G protein on a K+ channel, as shown in this Figure. Which one or more of the following would enhance this effect of acetylcholine? Explain. (a) A high concentration of a non-hydrolyzable analog of GTP. (b) Mutations in the acetylcholine receptor that weaken the interaction between the receptor and acetylcholine. (c) Mutations in the G protein α-subunit that speed-up the hydrolysis of GTP. (d) Mutations in the K+ Channel that make the βγ-subunit bind tighter Cancer and Cell Communication

17. How do activated G-proteins trigger release of ‘secondary messenger’ molecules? -- open channels -- activate enzymes Secondary messengers include: cAMP, Ca++, DAG, IP3 Some toxins interfere with G-proteins Cholera toxin Inhibits GTPase activity of α-subunit -- causes Na+ efflux into intestine -- water flow into intestine Pertussis toxin Prevents GDP/GTP exchange -- GTP locked in off state -- mucous secretion into lungs cAMP Signaling Cancer and Cell Communication

18. Downstream effects can be… Downstream enzyme activation (can be very rapid) -- effect of adrenaline Changes in gene expression (slower) There can be many other types of responses Block gene expression Activate exocytosis -- allergic responses -- insulin release or endocytosis -- phagocytic cells Cancer and Cell Communication

19. How do enzyme-linked receptors function? Receptor Tyrosine Kinases (RTK) Dimerization Autophosphorylation Activated signaling proteins Cancer and Cell Communication

20. In 25% of breast cancers, HER2 receptor is over-produced HER2/HER2 dimers form -- activated without hormone HER2 Signaling Cancer and Cell Communication

21. RTK Signaling often occurs through Ras A “monomeric” GTP-binding protein RAS activates a kinase “cascade” (MAP Kinase module) Cancer and Cell Communication

22. How are complex signaling pathways ‘dissected’? Genetically engineer cells to contain… -- Knockout mutations -- Constitutive expression mutations How do these 5 experiment establish signaling sequence of RAS, X and Y? Cancer and Cell Communication

23. When activated by the signal, the platelet-derived growth factor (PDGF) receptor phosphorylates itself on multiple tyrosines (as indicated below by the circled Ps; the numbers next to these Ps indicate the amino acid number of the tyrosine). These phosphorylated tyrosines serve as docking sites for proteins (A, B, C, and D) that interact with the activated PDGF-receptor. Binding of PDGF activates the PDGF-receptor leading to an increase in DNA synthesis. To determine whether protein A, B, C, and/or D are responsible for activation of DNA synthesis, you construct mutant versions of the PDGF-receptor that retain one or more tyrosine phosphorylation sites. In the cells, the various versions of the PDGF-receptor become phosphorylated on whichever tyrosines remain. You measure the level of DNA synthesis in cells that express the various mutant receptors and obtain the data shown below. • From these data, which, if any, of these proteins A, B, C, and D are involved in the stimulation of DNA synthesis by PDGF? Why? • Which, if any, of these proteins inhibit DNA synthesis? Why? • Which, if any, of these proteins appear to play no detectable role in DNA synthesis? Why? • What is the effect of the binding of A on the effect of B? Cancer and Cell Communication