special approaches of tumor biology and chemotaxis orsolya láng 2014
TRANSCRIPT
Special approaches of tumor biology and chemotaxis
Orsolya Láng
2014.
TUMOR CELLS AND MIGRATION
METASTASISPRIMARY TUMOR
AngiogenesisAdhesion
CELL and CELL CYCLE
Growth factors
Adhesion Adhesion moleculesmolecules
ChemokineChemokiness
Cell cyle Cell cyle regulatory regulatory proteinsproteins
Apoptotic Apoptotic moleculesmolecules
OncogenesOncogenes
TUMOR CELL
GENERAL FEATURE OF A TUMOR CELL
The hallmarks of cancer comprise six biological capabilities acquired during the multistep development of human tumors (Hanahan és Weinberg 2000)
CELL KINETICS
Doubling time of the tumor volume (Td)
Time of the Cell cycle (Tc):Tc= Ts / Li
Ts: S phaseLi: labeling index
(proportoin of cells in S phase)
Growth fraction(GF):GF=P / (P+Q’)
P: number of the mitotic cellsQ: number of the cells in interphase
Rate of the cell loss ():= 1-Tpd / Td Tpd= *Ts/Li
Tpd: Potential tumor volume doubling timeTd: tumor volume doubling time
Lymphoma 48 h
Lung cancer 108 h Usually 15-125 h
Lymphoma 4 weeksColon adenoma 90 weeksUsually 18-200 days
Volume of the tumor tissue
~10 divisions =*1000 cell number increase (210 =1024)~20 divisions= 106 cells = 1 mg= 1 mm3
~30 divisions= 109 cells = 1 g= 1 cm3
~40 divisions= 1012 cells = 1 kg
1 tumor cell ~30-33,25 division=1-10 cm3
Time of the clinical symptomes / diagnosis
~27 division= 0.1cm3
Earliest time of diagnosis(Visualisation)
~40 division= 1012cellFatal
Human total cell number: 3.72 × 1013
http://www.ncbi.nlm.nih.gov/pubmed/23829164
BIOLOGY OF THE TUMORPROGRESSION
Exogen and endogen factorsGenom instability
Activation of the oncogenesInactivation of tumorsuppressors
Local and systemic factorsinhibition acceleration
Growth rate
Ectopic survival capacity
Invasivity
De-differentation
Tumorigenesis
Epithelialcell
Hiperplasticadenoma
DisplaticCarcinoma in situ
Invasive carcinoma
Metastasis
Important steps of tumor progression
Transformation of the microenvironment: stromal cells,ECM components,proteolytic degradation
Induction of the angiogenesis (w/o max tumor size is 2mm)
Escaping from immune-mediated rejection Formation of metastasis
MICROENVIRONMENT – STROMAL CELLS
Cell types:fibroblasts,
myofibroblasts, endothelial cells, lymphocytes, macrophagesFunction:
host defence
! MALT - B cell helps to maintain lymphomas
! Growth factors are released by the stromal cells (VEGF - angiogenesis)
http://www.nature.com/nrc/journal/v9/n4/fig_tab/nrc2618_F1.html
Macrophage
Cancer cell migration is controlled by paracrine loop
THE INVASIVE MICROENVIRONMENT
http://www.nature.com/nrc/journal/v9/n4/full/nrc2618.html
Macrophage
Blood vessel
ANGIOGENESIS
Hypoxia formation of new vessels, proliferation of the endothelial cellsTypes: vessels
arteriovenous shunts„dead end”
/lack of smooth muscle , weak vessel wall, irregular shape(insuficient endothelial cell and basement membrane layers)/
sinuses /wall is formed by tumor cells/Venous circulation
VEGF induces angiogenesisincreases permeability
Lack of lymphatic vessels
OEDEMA, decresed blood flow
Inhibition of the VEGF pathway is a potential therapeutical tool
Strategies that tumors use to escape from immune-mediated rejection are:
To decrease the antigen expressionTo inhibit the immune-reactive cells:
degrade the chemoattractansdecrease their cell adhesioninhibite their phagocytotic activity
Angiogenesis
Local invasion
ECM
AdhesionProteolysisMigration
Intravasation
Extravasation
circulation
Metastasis
Tumor cell
Primary tumor
AdhesionProteolysisMigration
Angiogenesis
VEGFAngiogeni
nFGF
spreading
METASTATIC CASCADE
INVASION
In situ carcinoma
DECREASED CELL ADHESION, INCREASED MOTILITY
ECM proteolysis
Angiogenesis
Local invasion
ECM
AdhesionProteolysisMigration
Intravasation
Extravasation
circulation
Metastasis
Tumor cell
Primary tumor
AdhesionProteolysisMigration
Angiogenesis
VEGFAngiogeni
nFGF
spreading
METASTATIC CASCADE
CELL ADHESION
Significant change in cell-cell and cell-ECM interactionsMolecules:
selectinsintegrinsimmunoglobulin superfamily cadherins catenins
SELECTINS
Cell-cel junctionsTypes:
E- endothelial cellsP- trombocytesL- leukocytes
Extracellular C-lectin domain Ca2+ dependent anchorage It binds Sialyl-Lex carbohydrates
„ROLLING”
! Tumor cells express increased amount of sialil-Lex or -Lea
INTEGRINS
Transmembrane receptorsForm cell-ECM interaction8 , 14 subunites ~20 heterodimerCa2+, Mg2+ dependent anchorage„RGD” sequence is the specific substrateSignalling: outside-in – signalling
inside-out – adhesionIncreased expression of integrins promotes angiogenesis and helps to bind MMPs at the cell surfaceEXTRAVASATION, ATTACHMENT
D
G
R
Integrin or celladhesion regulated signalling pathways
cellproliferation
PTEN
RAC PI(3)K
CDC42
integrin
ECM
ILK
-cateninCiklin D1BAD
PKB/AKT
FAK
RASRAFMEKMAPK
GSK3
motilitygene expression
cellcycleapoptosis
SHCGRB2/SOS
Integrin or celladhesion regulated signalling pathways
integrin
ECM
ILK
-kateninCiklin D1BAD
PKB/AKT
FAK
cellproliferation
RASRAFMEKMAPK
GSK3
motilitygene expression
cellcycleapoptsis
SHCGRB2/SOS
PTEN
RAC PI(3)K
CDC42
Molecular partners of the integrins
Cytoskeletal components:actinin, talin,F- actin, filamin
Adaptors:rack 1, ICAP-1
Calcium binding proteins:CIB, calreticulin
Protein kinases:pp125FAK, p59 ILK
Membrane proteins:CD9, CD16,CD47…caveolin, urokinase-plazminogen-activator receptor
Ligands in ECM:collagen, laminin, fibronectin, fibrinogen, von
Willebrand factor, osteopontin, elastin
IMMUNGLOBULIN SUPERFAMILY
has 5 Ig-like domains at the extracellular regionforms cell-cell junction interacts with integrins
VCAM - 41, PECAM - v3
takes essential part in extravasation
! ! Over expression of ICAM-1, MUC18 increased inavsion
! ! Down-regulation of VCAM-1 increased metastatic potential (faster detachment)
CADHERIN
Is a transmembrane glycoproteinForms homophyl cell-cell junctionsCa2+ dependent anchorageClassical types: E- epithelial
P- placentaN- neural,
Intracellular part interacts with catenins to connect aktin filaments
! E-cadherin tumorsuppressor! Increased expression of N-cadherin invasion! N-cadherin cooperates with the FGF receptor lead to the up-
modulation of MMP-9
„Cadherin Switch”
CATENINCATENIN
Is an intracellular moleculeFixes cadherins to F-actin
! -catenin binds to the APC gene product
! colon and liver cancer increased cytoplasmic and nuclear localised beta-catenin correlates with invasion and poor prognosis.
in melanoma increased cytoplasmic and nuclear beta-catenin is currently emerging as a marker for good prognosis
Angiogenesis
Local invasion
ECM
AdhesionProteolysisMigration
Intravasation
Extravasation
Circulation
Metastasis
Tumor cell
Primary tumor
AdhesionProteolysisMigration
Angiogenesis
VEGFAngiogeni
nFGF
spreading
METASTATIC CASCADE
IntegrinscadherinsSelectins
CAM
PROTEOLYSISComponents of the basement membrane(BM) and ECM: IV collagen, laminin, proteoglycanesTumorcells (stromal cells) secrete proteasesCathepsinMatrix metalloproteinase (MMP)Plazmin, tPA ,Urokinase (plasminogen activator inhibitor 1&2)Tissue inhibitor of metalloproteinases
INVASION
MATRIX METTALLOPROTEINASESMATRIX METTALLOPROTEINASES (MMP)(MMP)
MOLECULAR STRUCTURE OF THE MATRIX MOLECULAR STRUCTURE OF THE MATRIX METTALLOPROTEINASESMETTALLOPROTEINASES
SUBSTRATE OF TIMP
Nature Reviews Cancer 2, 161-174 (March 2002)http://www.nature.com/nrc/journal/v2/n3/full/nrc745.html
MMP/TIMP EXPRESSION IN BREAST MMP/TIMP EXPRESSION IN BREAST CANCERCANCER
Nature Reviews Cancer 2, 161-174 (March 2002)
MMP – TUMORPROGRESSION?!?
Nature Reviews Cancer 2, 161-174 (March 2002)
Angiogenesis
Local invasion
ECM
AdhesionProteolysisMigration
Intravasation
Extravasation
Circulation
Metastasis
Tumor cell
Primary tumor
AdhesionProteolysisMigration
Angiogenesis
VEGFAngiogeni
nFGF
spreading
METASTATIC CASCADE
IntegrinscadherinsSelectins
CAM
MMP/TIMPCathepsin
Plasminogen
MIGRATORY MECHANISMS IN TUMOR
Small-cell lung cancer
FORMS OF MIGRATORY ADAPTATION
http://www.nature.com/nrc/journal/v3/n5/full/nrc1075.html
2D –3D MIGRATIONS
http://www.nature.com/nrc/journal/v3/n5/full/nrc1075.html
STEPS OF 3D MIGRATION
1. Pseudopod protrusion2. Formation of focal contact3. Focal ECM proteolysis4. Actomyosin contraction5. Detachment
Cell-cell interactions visualized in tumorigenesis
Angiogenesis
Local invasion
ECM
AdhesionProteolysisMigration
Intravasation
Extravasation
Circulation
Metastasis
Tumor cell
Primary tumor
AdhesionProteolysisMigration
Angiogenesis
VEGFAngiogeni
nFGF
spreading
METASTATIC CASCADE
IntegrinscadherinsSelectins
CAM
MMP/TIMPCathepsin
Plasminogen
AMF/gp78AutotaxinHGF/c-MET
!! Tumor markers e.g. cytokeratin, mucin
HEMATOGENIC DISSEMINATION
http://www.nature.com/nrc/journal/v4/n6/fig_tab/nrc1370_F3.html
Blood flow patterns can predict the specific regions of metastases in approximately two-thirds of cancers
TYPICAL SITE OF METASTASIS
LOCALISATION OF THE METASTASIS
CHEMOKINES – TISSUE SPECIFIC LOCALISATION
http://www.readcube.com/articles/10.1038/nrc865
adhesion
moti
lity
?
EXTRAVASATION
?
Attachment
Migration
http://www.nature.com/nrc/journal/v12/n1/fig_tab/nrc3180_F2.html#figure-title