Invasion and Metastasis: The Malignant Phenotype Folder Title: Inv&Mets(NoTP) Chapter 14: The Biology of Cancer Moving Out: Invasion and Metastasis p. 641 Second Edition Folder Title: Inv&Mets Malignancy is by defintion, the progressive emergence in a developing neoplasm of the ability to invade surrounding tissues and to spread to distant sites. It is these two properties of late-stage cancers that leads to their lethal outcomes. Thus understanding the biology of invasion and metastasis is key to understanding and managing malignant neoplasia. Updated: March 23, 2015

See Metastasis: Cancer Menacing Ballet (Insert by Robert Weinberg) by Jennifer Couzin (Insert by Robert Weinberg) Metastasis: Cancer's Menacing Ballet, (MetsScienceFeb1403.pdf) Science, Feb. 14, 2003, Vol 299, p 1003 Linked on “Password” Protected Site Course Web-page (No Password Needed)

Metastatic non-Hodgkins Lymphoma CT Scan and PET Scan (positron emission tomography) of incorporated radioactively-labelled deoxyfluoroglucose. (Brain activity is normal, abdominal active is pathological) Figure 14.1 The Biology of Cancer (© Garland Science 2007). P. 588; p. 642 2nd edition

Imaging on Metastatic Colon Carcinoma with Radioactive-Iodine-Labelled Monoclonal Ab to A33 Ag Lloyd Old, Scientific American, August, 1996, p. 138) SeeMets Arm

Colon Carcinoma Metastatic to Liver Fig. 2.2b and c Weinberg p. 27; p. 33 2nd edition Colon Carcinoma Metastatic to Liver Breast Carcinoma Metastatic to Brain

Multiple Metastatic Lesions of Gastric Adenocarcinoma to Liver See next slide

Gastric Carcinoma Metastatic to Brain

Primary Glioblastoma Compared to Breast Carcinoma Metastasis to the Brain

Growth-associated Neovascularization of a tumor xenograft Human colorectal adenocarcinoa implanted sub-cutaneously as a xenograft in immunocompromised mice. Viewed through a skin wndow. Figure 13.32a The Biology of Cancer (© Garland Science 2007) p. 561

Invasion-Metastasis Cascade Adapted from Fidler, Nat. Rev Invasion-Metastasis Cascade Adapted from Fidler, Nat. Rev. Cancer 3: 453-458, 2003 Figure 14.4 The Biology of Cancer (© Garland Science 2007) p. 591; Figure 14.4, p. 644, 2nd Edition

Breast Carcinoma Metastatic to Draining Lymph Node Figure 14.2b The Biology of Cancer (© Garland Science 2007). P 589

Carcinoma Metastatic to Bone. Stained for Epithelial Cell Markers Figure 14.2c The Biology of Cancer (© Garland Science 2007). P. 589

Turning Point Questions Coming Up Please get stuff off of the desks.

Invasion in Cancer

Cancer Invasion

Invasive Squamous Cell Carcinoma of Uterine Cervix Stromal Cells on Uterus Inflammatory Cells Invasive Carcinoma Figure 14.5c The Biology of Cancer (© Garland Science 2007) p. 592

Active Invasion by Melanoma Emboli Figure 14.5b The Biology of Cancer (© Garland Science 2007) p. 592

Detachment and Active Invasion by Renal Adenocarcinoma (Frog)

Progression in Invasion and Metastasis Appearance of Primary Tumor (Neoplasia in situ) Vascularization (Angiogenesis) Invasion Into surrounding tissue Into vascular and lymphatic systems Release of Tumor Emboli (Shedding) Systemic Transport Hematogenous Lymphatic Arrest at Distant Site Secondary Invasion: Extravasation Secondary Angiogenesis Secondary Invasion Tertiary Spread

Release of Tumor Emboli and Tumor Cell Shedding Invasion through thin anaplastic venous walls in tumor Facilitated by: Local trauma Diagnostic procedures Surgery Manipulation Emboli (small clumps of cells) Favored for survival by protection of inner cells Surrounded by fibrin clot May protect embolus while in circulation May facilitate survival of tumor cells at secondary tumor arrest site

Routes of Systemic Spread of Tumor Emboli and Tumor Cells Direct Extension Across Organ and Body Cavities Peritoneal Cavity Pleural Linings Peri-cardial Space Cerebrospinal Cavity Lymphatic Spread: Lymphatic capillaries to regional lymph nodes Hematogenous Spread: Entry via Lymphatic drainage into circulation Abnormal blood vessels in tumors Tumor cell deformability and motility

Arrest and Extravasation

Arrest of Tumor Emboli and Tumor Cells at Distant Sites Predilection for Specific Organ Sites Depends only partly on anatomical and circulatory relationships Specific Organ Homing Based on Cell Adhesion Recognition Cell-Cell and Cell-Connective Tissue Adherence Plasma membrane ligands on metastatic tumor cells Cell adhesion receptors on endothelial lining of capillaries in target organs Binding to laminin and fibronectin in extra-cellular connective tissue matrix

Extravasation Facilitated by Clot Formation Figure 14.9 The Biology of Cancer (© Garland Science 2007) p. 595

Epithelial to Mesenchymal Transition in Cancer Detachment and Invasion Mesenchymal to Epithelial Transition in Establishing Disseminated Metastases

Association of Normal Melanocyte with Epithelial Keratinocytes (Epithelial adherens junction protein) p. 605; p. 662 2nd edition Figure 14.16a The Biology of Cancer (© Garland Science 2007)

Epithelial to Mesenchymal Transition in Melanoma Cells: Facilitation of Detachment and Invasion (Mesenchymal adherens junction protein) p. 605; p. 662 2nd edition Figure 14.16b The Biology of Cancer (© Garland Science 2007)

Epithelial-Mesenchymal Transition Non-motile Epithelial Cells Associated with Each Other via E-Cadherin Cell Surface Attachment Receptor Anchored to Connective Tissue Basement Membrane by E-Cadherin Tight Association via E-Cadherin Express Intermediate Filament Protein Cytokeratin: Characteristic of Epithelial Cells. Invasive Carcinoma Cells: Morphology and Gene-expression Converted to Connective Tissue Type Cells Express N-Cadherin: Loosely and Reversibly Associated with Each Other and with Connective Tissue Express Intermediate Filament Protein Vimentin: Characteristic of Connective Tissue Cells Fibroblast and Leucocyte-like Structure and Function Able to Migrate and to Cross Circulatory and Connective Tissue Barriers Re-use Gene Expression and Functions from Embryonic and Wound-healing States Revert back to Epithelial Characteristics after Seeding Distant Site: “Mesenchymal-Epithelial Transition”

p. 603 Table 14.2 The Biology of Cancer (© Garland Science 2007)

Reversibility of Epithelial-Mesenchymal Transition: To Invasive Carcinoma and Back to Macrometastasis at Distant Site p. 606; p. 665, 2nd edition Figure 14.17b The Biology of Cancer (© Garland Science 2007)

Reversibility of Epithelial – Mesenchymal Transition: Epithelial Characteristics of Distant Metastases of Primary Carcinoma (Aberrant epidermal growth factor receptor) p. 607 Figure 14.18 The Biology of Cancer (© Garland Science 2007)

Most Frequent Sites of Metastases for Some Human Cancer Axillary lymph nodes, other breast, lung, pleura, liver, bone brain, spleen, adrenals, ovary Regional lymph nodes, liver, lung, bladder, stomach Lung, liver, bone Regional lymph nodes, pleura, diaphagm, liver, bone, brain, kidney, adrenal, throid, spleen Peritoneum, regional lymph nodes, lung, liver Bones of spine and pelvis, regional lymph nodes Regional lymph nodes, liver, lung, bone Regional lymph nodes, lung. liver Rectum, colon, prostate, ureter, vagina, bone, regional lymph nodes, lung, peritoneum, pleura, liver, brain Regional lymph nodes, lung, liver, ovary Breast Colon Kidney Lung Ovary Prostate Stomach Testis Urinary Bladder Uterine Lining

Primary Tumors and Preferred Sites of Metastatic Spread Figure 14.42 The Biology of Cancer (© Garland Science 2007)

Presence of Micrometastases and Clinical Prognosis: Breast Cancer Figure 14.50a The Biology of Cancer (© Garland Science 2007)

Presence of Micrometastases and Clinical Prognosis: Colon Cancer Figure 14.50b The Biology of Cancer (© Garland Science 2007) p. 645

Over- or Under-expression of 128 Metastasis-Associated Genes in DNA-Array Assay for Potential Metastatic Progression. RNA’s prepared from 64 primary non-metastatic adenocarcinomas, and from 12 metastatic adenocarcinomas (17 representative Genes) . . Blue means decreased expression. Half of the 128 genes in the 12 mets were down-regulated. Red means increased expression. Half were up-regulated. Figure 14.51a The Biology of Cancer (© Garland Science 2007) p. 714, Fig. 14.51, 2nd Edition

p. 647; See page 714, 2nd edition Figure 14.51b The Biology of Cancer (© Garland Science 2007)

Turning Point Questions Coming Up Please get stuff off of the desks.

Size of Primary Breast Cancer and Risk of Metastasis: 46-Year Follow-up (Figure 14.3, p. 590 Figure 14.3 The Biology of Cancer (© Garland Science 2007)

Secondary Metastatic Growth Growth at site of secondary arrest Protection by fibrin clot? Secondary Invasion Out of vasculature into target tissue Active Passive Growth of Metastatic Nodules Angiogenesis Invasion into metastatic organ site Potential for Tertiary Invasion

Extravasation Facilitated by Clot Formation Figure 14.9 The Biology of Cancer (© Garland Science 2007) p. 595

Reversibility of Epithelial-Mesenchymal Transition: To Invasive Carcinoma and Back to Macrometastasis at Distant Site p. 606; p. 665, 2nd edition Figure 14.17b The Biology of Cancer (© Garland Science 2007)

Factors Contributing to Metastatic Spread Metastasis-Associated Up-regulated Genes: Promotion of Epitelial-Mesenchymal Transition Host Responses (not necessarily immunological) Inflammatory responses: See Macrophages and Promotion of Metastasis, Figures 14.22 and 14.23, pp. 612-613 At Primary Site At Potential Seeding Site. See Scientific American, March 2007 Article: “Deadly Dialogue”. (Not required reading but of value now or later) Clot Formation Cytokine and Growth Factor Production Tumor Responses Tumor-induced immune suppression Possible Facilitation of Metastasis by Treatment Diagnostic and surgical manipulation X-ray Damage Immune suppression by Drug Treatment by Surgery and Anesthesia by Stress Hormones

Reciprocal Stimulation of Breast Cancer Cells by Macrophages: Stimulation of Proliferation and Migration of Carcinoma Cells by Epidermal Growth Factor (EGF) from Macrophages p. 673, 2nd edition

EGF = Epidermal Growth Factor CSF-1 = Colony-Stimulating Factor Reciprocal Stimulation of Breast Cancer Cells & Macrophages: Recruitment of Macrophages by Colony Stimulating Factor from Cancer Cells, and Promotion of Entry into Vasculature by Inflammatory Macrophages Tumor-associated Macrophage EGF = Epidermal Growth Factor CSF-1 = Colony-Stimulating Factor p. 673, 2nd edition

Factors Hindering Metastatic Spread Metastasis-Suppressor Genes: See Table 14.4, p. 643 e.g. TIMP: Tissue Inhibitor of Metalloproteinases or RhoGD1-2: Down-regulates Rho – Stimulator of Actin Polymerization Host Responses Activated Macrophages Natural Killer Cells Cytotoxic Lymphocytes Hydrodynamic Effects in Host circulation Failure to Recognize and Arrest at Secondary Site