Robert A.

Weinberg

The Biology of Cancer

First Edition

Chapter 14: Moving Out: Invasion and Metastasis

Copyright Garland Science 2007

CT-PET fusion image of a lymphoma patient

Figure 14.1 The Biology of Cancer ( Garland Science 2007)

Pancreas of Rip-Tag transgenic mouse (transgene of SV40 large T and small T antigens)

The Rip-Tag model of islet cell tumor progression

Figure 13.37 The Biology of Cancer ( Garland Science 2007)

Metastatic islet cell tumor in a lymphatic channels

Endothelial cells

Figure 14.2a The Biology of Cancer ( Garland Science 2007)

Breast cancer cells in lymph node

Figure 14.2b The Biology of Cancer ( Garland Science 2007)

Metastatic cancer cells in bone marrow (Wright-Giemsa stain

Figure 14.2c The Biology of Cancer ( Garland Science 2007)

Section 1.

Travel of cancer cells from a primary tumor to a site of potential metastasis depends on a series of complex biological steps

Figure 14.3 The Biology of Cancer ( Garland Science 2007)

The structure of basement membrane (specialized ECM) Hemidesmosome is composed of integrin and laminin

Figure 13.5a The Biology of Cancer ( Garland Science 2007)

The structure of basement membrane Extra-cellular Matrix (ECM) protein

Figure 13.5b The Biology of Cancer ( Garland Science 2007)

The invasion metastasis cascade: six steps are involved

Figure 14.4 The Biology of Cancer ( Garland Science 2007)

Patterns of Invasion

Lobular carcinoma of breast

Figure 14.5a The Biology of Cancer ( Garland Science 2007)

Red: E-cadherin, green: beta-1-integrin, blue: matrix

Melanoma cells

Figure 14.5b The Biology of Cancer ( Garland Science 2007)

Squamous cell carcinoma of cervix

Figure 14.5c The Biology of Cancer ( Garland Science 2007)

Figure 13.39b The Biology of Cancer ( Garland Science 2007)

Figure 13.39a The Biology of Cancer ( Garland Science 2007)

Figure 14.6 The Biology of Cancer ( Garland Science 2007)

Intra-vital microscopy

Figure 14.7a The Biology of Cancer ( Garland Science 2007)

Intravital fluorescence microscopy

Green: plasma Red: RBC

Figure 14.7b The Biology of Cancer ( Garland Science 2007)

Confocal microscopy Green: rat fibrosarcoma cell Red: LDL of the arteriole

5 days after injection of single cells injection into mouse vein

Figure 14.8 The Biology of Cancer ( Garland Science 2007)

Extravasation

Diapedesis
Cancer cell push aside the endothelial cells, reach basement membrane

Resolving of the thrombus

Figure 14.9 The Biology of Cancer ( Garland Science 2007)

Section 2. Colonization represents the most complex and challenging step of the invasion-metastasis cascade

Micro-metastasis in bone marrow: colon cancer (anti-cytokeratin antibody stain)

Figure 14.10a The Biology of Cancer ( Garland Science 2007)

Micro-metastasis in bone marrow: breast cancer

Figure 14.10b The Biology of Cancer ( Garland Science 2007)

Two clusters of lung adenocarcinoma in the lymph node

Figure 14.10c The Biology of Cancer ( Garland Science 2007)

Genetic heterogeneity of micrometastasis and the evolution of colonizing ability

Figure 14.11a, b The Biology of Cancer ( Garland Science 2007)

Figure 14.11c The Biology of Cancer ( Garland Science 2007)

Persistence of solitary dormant () tumor cells many weeks after introduction into the liver

Following isolation and in vitro culturing, the descendants of many of these cells were tumorigenic
Figure 14.12 The Biology of Cancer ( Garland Science 2007)

Section 3. The epithelial-mesenchymal transition and associated loss of Ecadherin expression enable carcinoma cells to become invasive

Embryogenesis and the epithelial-mesenchymal transition (EMT)

Sea urchin embryo

Figure 14.13a The Biology of Cancer ( Garland Science 2007)

Figure 14.13b The Biology of Cancer ( Garland Science 2007)

Table 14.1 The Biology of Cancer ( Garland Science 2007)

The program of wound healing

Figure 13.14 The Biology of Cancer ( Garland Science 2007)

EMT was provoked by a 3-day-long exposure to MMP-3 , which

could initiate EMT through its ability to degrade E-cadherin.

red: cytokeratin, green: vimentin

Figure 13.13a The Biology of Cancer ( Garland Science 2007)

Individual cells may spontaneously have EMT, suggesting the plasticity of these cells.

Green: actin Red: cytokeratin

Figure 13.13b The Biology of Cancer ( Garland Science 2007)

The serial changes of a mono-layer of breast epithelial cells after a patch of cells were removed.

Figure 13.13c The Biology of Cancer ( Garland Science 2007)

Table 14.2 The Biology of Cancer ( Garland Science 2007)

EMT at the invasive edge of a malignant tumor

Immunohistochemical stain for E-cadherin

Figure 14.14a The Biology of Cancer ( Garland Science 2007)

Immunohistochemical stain for -catenin

Figure 14.14b The Biology of Cancer ( Garland Science 2007)

Figure 14.14c The Biology of Cancer ( Garland Science 2007)

Figure 13.12a The Biology of Cancer ( Garland Science 2007)

Figure 13.12b The Biology of Cancer ( Garland Science 2007)

Figure 13.12c The Biology of Cancer ( Garland Science 2007)

Immunofluorescence stain of Keratinocytes Yellow: E-cadherin, red: actin

Figure 13.12d The Biology of Cancer ( Garland Science 2007)

Figure 6.26a The Biology of Cancer ( Garland Science 2007)

Figure 14.16a The Biology of Cancer ( Garland Science 2007)

Figure 14.16b The Biology of Cancer ( Garland Science 2007)

Table 7.1 part 1 of 2 The Biology of Cancer ( Garland Science 2007)

Table 7.1 part 2 of 2 The Biology of Cancer ( Garland Science 2007)

Table 7.2 The Biology of Cancer ( Garland Science 2007)

The EMT can be induced by several transcriptional factors Example: Twist on canine kidney cells

Figure 14.15a The Biology of Cancer ( Garland Science 2007)

Figure 14.15b The Biology of Cancer ( Garland Science 2007)

Section 4. The epithelial-mesenchymal transition is often induced by stromal signals

Reversibility of EMT: Release of degradative enzymes, such as MMPs, is noted in EMT. EMT may be triggered by the signals from the tumor associated stroma

Red: CK18 Green: basement membrane

Figure 14.17a The Biology of Cancer ( Garland Science 2007)

Figure 14.17b The Biology of Cancer ( Garland Science 2007)

The reversibility of EMT explains the peculiarity of many metastasis: they are similar to the primary tumor

Figure 14.18 The Biology of Cancer ( Garland Science 2007)

Manifestation of EMT at the interface between tumor epithelium and stroma

Figure 14.19a The Biology of Cancer ( Garland Science 2007)

Laminin-2

Figure 14.19b The Biology of Cancer ( Garland Science 2007)

Figure 14.19c The Biology of Cancer ( Garland Science 2007)

Vimentin

Figure 14.19d The Biology of Cancer ( Garland Science 2007)

Abudant evidence indicates that TGF- is an important agent for conveying these stromal signals

Figure 14.20a The Biology of Cancer ( Garland Science 2007)

Control of the EMT by TGF- and its effect on tumorigenic cells

Figure 14.20b The Biology of Cancer ( Garland Science 2007)

Figure 14.20c The Biology of Cancer ( Garland Science 2007)

Figure 14.20d The Biology of Cancer ( Garland Science 2007)

Figure 14.20e The Biology of Cancer ( Garland Science 2007)

TNF- and TGF- contribute to active NF-kB signaling

NF-kB signaling was necessary for EMT in the EpRas cell line
Figure 14.21 The Biology of Cancer ( Garland Science 2007)

The effect of stromal macrophages on the invasive and metastatic behavior of cancer cells
Transgenic mice model

TAM(+)

TAM (-)

Colony stimulating factor -1 (CSF-1) for recruit tumor associated macrophages (TAM)
Figure 13.24 The Biology of Cancer ( Garland Science 2007)

Colony stimulating factor -1 (CSF-1) for recruit tumor associated macrophages (TAM)

Figure 14.22a The Biology of Cancer ( Garland Science 2007)

Figure 14.22b The Biology of Cancer ( Garland Science 2007)

Metastasis in the lung increase with age in the CSF-1(+) mice

Figure 14.22c The Biology of Cancer ( Garland Science 2007)

Figure 14.23a The Biology of Cancer ( Garland Science 2007)

Figure 14.23b The Biology of Cancer ( Garland Science 2007)

Since macrophages are often found in close to microvessels, the stimulation by tumor associated macrophages (TAM) may also contribute to cancer cell intravasation

Figure 14.23c The Biology of Cancer ( Garland Science 2007)

Cell scattering and invasive behavior induced by hepatocyte growth factor (HGF) or SF

HGF is another ligand of stromal origin, which is also capable of inducing EMT in the epithelial cells trough the effect of Met protein (HGF receptor)
Figure 14.24 The Biology of Cancer ( Garland Science 2007)

Signals that triggered EMT

Figure 14.25 The Biology of Cancer ( Garland Science 2007)

Section 5. EMTs are programmed by transcription factors that orchestrate key steps of embryogenesis

Table 14.3 The Biology of Cancer ( Garland Science 2007)

Embryonic transcriptional factors programing EMT

Figure 14.26a The Biology of Cancer ( Garland Science 2007)

Figure 14.26b The Biology of Cancer ( Garland Science 2007)

Figure 14.26c The Biology of Cancer ( Garland Science 2007)

Figure 14.26d The Biology of Cancer ( Garland Science 2007)

Figure 14.26e The Biology of Cancer ( Garland Science 2007)

Figure 14.26f The Biology of Cancer ( Garland Science 2007)

Slug transcription factor in wound healing

Figure 14.27 The Biology of Cancer ( Garland Science 2007)

Expression of EMT-inducing embryonic transcription factors in human tumors

Slug suppress E-cadherin transcription

Figure 14.28a The Biology of Cancer ( Garland Science 2007)

Expression of EMT-inducing embryonic transcription factors in human tumors

Figure 14.28b The Biology of Cancer ( Garland Science 2007)

Expression of EMT-inducing embryonic transcription factors in human tumors

Figure 14.28c The Biology of Cancer ( Garland Science 2007)

Embryonic transcription factors and tumor progression

Figure 14.29a The Biology of Cancer ( Garland Science 2007)

Embryonic transcription factors and tumor progression

Figure 14.29b The Biology of Cancer ( Garland Science 2007)

Similarities between EMT signaling during embryogenesis and tumor progression

Figure 14.30 The Biology of Cancer ( Garland Science 2007)

Mammary carcinoma arising from transgenic MMTV-polyoma T mice

Matrix metalloproteinases (MMPs) produced by tumor associated cells, would generate a halo of proteolysis (right figure)
Figure 14.31a The Biology of Cancer ( Garland Science 2007)

The ability of tumor cells to degrade collagen IV fibers trough interaction with fibroblasts

Red: fibroblast Green: degraded collagen IV

Figure 14.31b The Biology of Cancer ( Garland Science 2007)

Macrophages are important source of MMPs

Elevated MMP-2 imparted increased invasiveness to the breast cancer cells

Figure 14.31c The Biology of Cancer ( Garland Science 2007)

Section 6. Extracellular proteases play key roles in invasiveness

Table 13.1 The Biology of Cancer ( Garland Science 2007)

Tumorassociated macrophages

Figure 13.25a The Biology of Cancer ( Garland Science 2007)

VEGF expression in tumor cells

VEGF expression in TAM

Figure 13.25b The Biology of Cancer ( Garland Science 2007)

Figure 13.25c The Biology of Cancer ( Garland Science 2007)

TAM with expression of MMP-9, a key enzyme in angiogenesis and invasiveness.

Figure 13.25d The Biology of Cancer ( Garland Science 2007)

Contribution of macrophages to tumorigenesis

Figure 13.26 The Biology of Cancer ( Garland Science 2007)

Podosomes are small, focal protrusions from the cell surface, that are used to degrade the extracellular matrix (ECM) of immediate vicinity. In cancer cells, they can also be called as invadosomes

Red: actin, green: ECM protein

Figure 14.32 The Biology of Cancer ( Garland Science 2007)

Ectopic expression of MMP-3 and mammary tumor progression

Figure 14.33 The Biology of Cancer ( Garland Science 2007)

Activation of extracellular proteins: MMPs and TGF- by uPA and uPAR

uPA: urokinase plasminogen activator

Figure 14.34 The Biology of Cancer ( Garland Science 2007)

Section 7. Small Ras-like GTPases control cellular processes including adhesion,cell shape,and cell motility

Figure 14.35 The Biology of Cancer ( Garland Science 2007)

Lamellipodia of rat liver cells

Figure 14.36a The Biology of Cancer ( Garland Science 2007)

Lamellipodia of a fibroblast Red: actin Green:Ena protein

Figure 14.36b The Biology of Cancer ( Garland Science 2007)

Fish keratinocytes with prominent lamellipodia. EM demonstrate densely woven network of actin filaments to extend the lamellipodia in the direction of movement.

Figure 14.36c The Biology of Cancer ( Garland Science 2007)

The addition of heregulin induce the development of lamellipodia that faces all directions

Breast cancer cells

Green: actin

Figure 14.36d The Biology of Cancer ( Garland Science 2007)

Filopodia

Figure 14.37a The Biology of Cancer ( Garland Science 2007)

The leading edges of lamellipodia are ofteninterspersed with filopodia

Figure 14.37b The Biology of Cancer ( Garland Science 2007)

The actin fibers within a filopodium

Figure 14.37c The Biology of Cancer ( Garland Science 2007)

Effects of Rho-like proteins on the actin cytoskeleton and cell adhesion

Figure 14.38 The Biology of Cancer ( Garland Science 2007)

Figure 14.38a The Biology of Cancer ( Garland Science 2007)

Figure 14.38b The Biology of Cancer ( Garland Science 2007)

Figure 14.38c The Biology of Cancer ( Garland Science 2007)

Figure 14.38d The Biology of Cancer ( Garland Science 2007)

The circuitry mediating EGFinduced cell motility

Figure 14.39 The Biology of Cancer ( Garland Science 2007)

Influence of RhoC on metastasis

Figure 14.40 The Biology of Cancer ( Garland Science 2007)

Section 8. Metastasizing cells can use lymphatic vessels to disperse from the primary tumor

Draining lymph nodes of the mammary glands

Figure 14.41a The Biology of Cancer ( Garland Science 2007)

Detection of sentinal lymph node by injection of a blue dye

Figure 14.41b The Biology of Cancer ( Garland Science 2007)

Figure 14.41c The Biology of Cancer ( Garland Science 2007)

Figure 14.41d The Biology of Cancer ( Garland Science 2007)

Section 9. A variety of factors govern the organ sites in which disseminated cancer cells form metastases

Primary tumors and their metastatic tropisms

Figure 14.42 The Biology of Cancer ( Garland Science 2007)

Figure 14.44 The Biology of Cancer ( Garland Science 2007)

Section 10. Metastasis to bone requires the subversion of osteoblasts and osteoclasts

Bone degradation by osteoclasts

Figure 14.45a The Biology of Cancer ( Garland Science 2007)

An osteoclast has excavated a shallow pit, which revealed the complex meshwork of the bone matrix.

Figure 14.45b The Biology of Cancer ( Garland Science 2007)

Osteolytic lesion by tumor metastasis (scanning EM)

Figure 14.45c The Biology of Cancer ( Garland Science 2007)

Figure 14.46 The Biology of Cancer ( Garland Science 2007)

The physiologic balance between bone formation and resorption

osteoprotegerin

Figure 14.47 The Biology of Cancer ( Garland Science 2007)

The vicious cycle of osteolytic metastasis

The bone matrix is a rich source of mitogenic and trophic factors

Parathyroid hormone related peptides

Figure 14.48 The Biology of Cancer ( Garland Science 2007)

MDA-MB-231is a breast cancer cell line usually produce osteolytic metastasis.

TGF- stimulates osteolytic activity by forcing the breast cancer cells to release PTHrP, which activate osteoblasts, then--- osteoclasts.
Figure 14.49 The Biology of Cancer ( Garland Science 2007)

Section 11. Metastasis suppressor genes contribute to regulating the metastatic phenotype

Table 14.4 The Biology of Cancer ( Garland Science 2007)

Section 12. Occult micrometastases threaten the long-term survival of cancer patients

Figure 14.50a The Biology of Cancer ( Garland Science 2007)

Figure 14.50b The Biology of Cancer ( Garland Science 2007)

Use of expression array to predic disease progression

Figure 14.51a The Biology of Cancer ( Garland Science 2007)

Figure 14.51b The Biology of Cancer ( Garland Science 2007)

Gene similarity between primary tumors and derived metastasis

Figure 14.52a The Biology of Cancer ( Garland Science 2007)

Figure 14.52b The Biology of Cancer ( Garland Science 2007)

Some expressed genes within tumor cells facilitate specific types of metastasis

Figure 14.53 The Biology of Cancer ( Garland Science 2007)

The End