1. Invasion and Metastasis: The Malignant Phenotype Updated: March 26, 2014 Folder Title: Inv&Mets Chapter 14: The Biology of Cancer Moving Out: Invasion and Metastasis p. 641 Second Edition

2. See Metastasis: Cancer Menacing Ballet by Jennifer Couzin (Insert by Robert Weinberg) Metastasis: Cancer's Menacing Ballet,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)

3. Figure 14.1 The Biology of Cancer (© Garland Science 2007). P. 588 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)

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

5. Colon Carcinoma Metastatic to Liver Breast Carcinoma Metastatic to Brain Fig. 2.2b and c Weinberg p. 27

6. Multiple Metastatic Lesion to Liver

7. Gastric Carcinoma Metastatic to Brain

8. Primary Glioblastoma Compared to Breast Carcinoma Metastasis to the Brain

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

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

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

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

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

17. In the previous slide breast carcinoma metastatic to the bone was detected by using stains for epithelial cell markers. Why does that work? RankResponses 1 2 3 4 5 6Other

18. Angiogenesis can promote intravasation. Why do you think that happens? RankResponses 1 2 3 4 5 6Other

19. Invasion in Cancer

20. Cancer Invasion

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

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

23. Detachment and Active Invasion by RenalDetachment and Active Invasion by Renal Detachment and Active Invasion by Renal Adenocarcinoma (Frog)

24. 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

25. 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

26. 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

27. Arrest and Extravasation

28. 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

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

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

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

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

33. 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”

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

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

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

37. Most Frequent Sites of Metastases for Some Human Cancer Breast Colon Kidney Lung Ovary Prostate Stomach Testis Urinary Bladder Uterine Lining 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

38. Figure 14.42 The Biology of Cancer (© Garland Science 2007) p. 635 Primary Tumors and Preferred Sites of Metastatic Spread

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

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

41. Figure 14.51a The Biology of Cancer (© Garland Science 2007) p. 647 Over- or Under-expression of 128 Metastasis-Associated Genes in DNA-Array Assay for Potential Metastatic Progression

42. Figure 14.51b The Biology of Cancer (© Garland Science 2007) p. 647

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

44. Primary localized carcinoma cells can become invasive and metastatic by invoking genes, gene products, and functions of mesenchymal connective tissue type cells (the epithelial-mesenchymal-transition). Name any one feature, molecule, function, or property of mesenchymal cells that could help to promote invasion and metastasis. RankResponses 1 2 3 4 5 6Other

45. The blue line and the red line in the medulloblastoma cancer (panel III) run very close together after 90 months. What does that tell you about the primary medulloblastoma with no metastasis signature?

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

47. 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

48. 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

49. Reciprocal Stimulation of Breast Cancer Cells by Macrophages: Stimulation of Proliferation and Migration of Carcinoma Cells by Epidermal Growth Factor (EGF) from Macrophages

50. 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

51. 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