Cell Cycle, Growth Factors, Neoplasia (Oncogenesis)

Definitions
Normal homeostasis (Balance of proliferation and Apoptosis)
• Regeneration – growth of cells and tissues to replace lost structures. Requires
intact connective tissue scaffolding of ECM:
• Healing
• Response to a wound, chronic inflammation and cell necrosis in organs
incapable of regeneration.
• Balance of variable properties of regeneration and scar formation.
Control of Normal Cell Proliferation and Tissue Growth
• Cell population size depends on rate of proliferation, differentiation and death by
apoptosis.
Apoptosis – Physiological process required for tissue homeostasis may be pathological.
• Tissue Proliferative Activity
• Cell cycle consists of G1 (pre-synthetic), S (DNA synthesis), G2 (pre-mitotic and
M (mitotic) phases.
• Go = Quiescent Cells
• Labile – continuously dividing and cycling. Require Stem Cell
• Quiescent – Normal has a (stable) low level of replication but can be stimulated
to undergo rapid division. In Go stage.
• Permanent – Non dividing left cycle and cannot undergo mitotic activity in post
natal life.
GROWTH FACTORS
• Polypeptides which may act on many cells or have restricted cellular targets
• Stimulate cell proliferation
• Affect contractility, differentiation and angiogenesis, tissue regeneration and
repair.
Growth Factors in Regeneration and Repair
• Epidermal Growth Factor (EGF) and Transforming Growth Factor & (TGFX)
• Belong to family of EGF with common receptor
• Mitogenic to keratinocyte migration and formation of granulation tissue
• TGF similar to EGF and stimulates hepatocyte proliferation. Oncogenic
• Produced by platelets, macrophages, T lymphocytes keratinocytes
• “EGF receptor” a family of membrane tyrosine kinase receptors which respond to
EGF, TGFX
• Main EGFR is EGFR1 or ERB B1
• ERB B2 (HER2) over-expressed in breast carcinomas
Hepatocyte Growth Factor (HGF)
• Proliferation of epithelial, endothelial cells and hepatocytes
• cell motility
• Promotes cell migration (i.e. morphogen)
• Produced by mesenchymal cells (fibroblasts, endothelial cells, liver
mesenchymal cells)
• Receptor is product of proto-oncogene C.MET often over-expressed in human
tumours
Vascular Endothelial Growth Factor (VEGF)
• Produced by mesenchymal cells
• Isoforms A,B,C,D
• Mitogenic for endothelial cells, high vascular permeability
• Signal through tyrosine kinase receptors
Platelet Derived Growth Factor (PDGF)
• Isoforms A; B; C; D
• Sources – Platelets, macrophages, endothelial cels, smooth muscle cells and
keratinocytes
• Chemotactic for PMNS macrophages, fibroblasts, smooth muscle cells and
activates them
• Mitogenic for fibroblasts, smooth muscle cells and endothelial cells
• Stimulates angiogenesis and wound contraction
• Inhibit platelet aggregation
• Bind to cell – surface receptors PDGF a and b
Fibroblast Growth Factor (FGF)
• Produced by macrophages, mast cells, fibroblasts and endothelial cells.
• Chemotactic for fibroblasts
• Mitogenic for fibroblasts and keratinocytes
• Angiogenesis, wound repair
• Development of lungs and skeletal muscle
Transforming Growth Factor (TGFB)
• Isoforms 1,2,3
• Produced by T lymphocytes, macrophages, endothelial cells, keratinocytes,
fibroblasts, smooth muscle cells, platelets
• Chemotactic for PMN’S macrophages, fibroblasts, smooth muscle cells
• Angiogenesis and fibroplastic
Insulin-like Growth Factor (IGF-1)
• Produced by fibroblasts, etc.
• Stimulates synthesis of collagen and fibroblast proliferation
• Endocrine effects similar to growth hormone
Tumour Necrosis Factor (TNF)
• Produced by macrophages, T cells and mast cells
• Activates macrophages
Interleukins (IL-I) etc
• Macrophages, mast cells, lymphocytes etc.
• Multiple functions including chemotaxis, angiogenesis etc.
Interferons (IFNa) etc
• Produced by lymphocytes, fibroblasts
• Activates macrophages, regulates other cytokines
• Ligand – receptor binding triggers events to extracellular signal transduction into
cell and modulation of gene expression
• Common form involves receptor di / trimerisation
 • Single receptors require cytoplasmic adapter or bridging proteins
• Receptors occur on cell surface, cytoplasm an nucleus
Major Types of Receptors
• - Mechanisms of signal transduction

A. Receptors with intrinsic Tyrosine kinase Activity

- Ligands for these receptors include growth factors (EGF, TGF - a , HGF, VEGF,
FGF, CKIT, Insulin
- They have extrocellular domain (ligand-binding) transmembrane and a
cytoplasmic tail with intrinsic tyrosine kinase activity
- Binding causes dimerisation of receptor, tyrosine phosphorylation and activation
of receptor tyrosine kinase
- Activate kinase phosphorylates and activates downstream effector molecules
Phosphorylated residues in receptor serve as docking sites for adapter molecules (SOS,
GRB2) that bind effector molecules
- Effector molecules include phospholipase (PLCV), PI3 kinase
- PI3 kinase phosphorylates membrane phospholipid
- GRB2 (adapter protein) binds a GTP binding (G) protein RAS
- MAD = mitogen activated protein kinase
B. Receptors without intrinsic T.K. activity that recruit kinases
- Ligands include cytokines (IL-2, IL-3), interferons growth hormone, prolactin
- Receptors activate JAK (Janus Kinase) family of proteins and activate STATS
(Signal transduces and activation of transcription) which shuffle into nucleus and
activate transcription
- Cytokine – receptors can activate other signaling pathways e.g. MAP kinase
C. Seven transmembrane G protein coupled receptors “GPCR’S”
• Contain 7 transmembrane µ helices
• Largest family of plasma membrane receptors
• Transmit via trimeric GTP – binding proteins (G proteins)
• Ligands include serotonin adrenaline, glucagon
• Binding causes receptor conformation which interact with G proteins
• Involve CAMP and CA++ as second messengers
D. Steroid hormone receptors
• Ligands for these receptors diffuse through cell membrane
• Bind to receptors in nucleus or cytoplasm
• Receptors are transcript on factors that activate transcription
• e.g. Oestrogen receptor localised in cytoplasm
Transcription Factors
• Transfer information to nucleus and modutate gene transcription
• TF’s include products of several growth promoting genes e.g. c-myc, cjun and
p53
Cell Cycle and Regulation of Cell Replication
• Replication of cells directly stimulated by growth factors or by signaling from
ECM components through intergrins
• Cell cycle stops at sites where essential gene function is deficient
• Cell cycle has multiple controls especially during transition between G1 and S
phases i.e. activators, inhibitors and checkpoint mechanisms
• To enter cycle quiescent cells must go through transition from Go to G1 which
requires transcriptional activation of genes including proto-oncogenes
• G1/S transition is a restriction point and rate limiting for replication
• Progression through cell cycle (esp G1/S) controlled by proteins called cyclins
and (CDKs) cyclin dependant kinases
• CDKs bind to cyclins and acquire catalytic activity
• Activated CDK’s drive cycle by phosphorylating proteins critical cycle transitions
• Activity of cyclin-CDK regulated by inhibitors (CDK inhibitors)
• Some growth factors inhibit products of CDKI’s
• Checkpoints are surveillance mechanisms for sensing DNA and chromosomal
damage and preventing such cells from completing replication e.g. G1/S
checkpoint for DNA damage before and G2/M after replication
• Checkpoint delay replication and allow time for repair mechanisms
• If damage too severe for repair, apoptosis initiated via p53 dependent
mechanisms