1. Student Learning Outcomes:
18 Cancer
Chapter 18 Cancer:
Student Learning Outcomes:
Explain causes, development of cancer (clonal):
Chemicals, radiation, virus, spontaneous
Decribe diversity of tumor viruses (model systems)
Explain essential features of Oncogenes:
Derived from proto-oncogenes, examples
Explain essential features of tumor suppressors:
Normal cell functions, examples
Describe molecular approaches to cancer treatment

2. Introduction
18.1 Cancer cells have abnormalities in multiple cell regulatory systems.
Breakdown of regulatory mechanisms that govern normal cell behavior:
Grow, divide in uncontrolled manner,
Spread throughout body
Interfere with function of normal tissues, organs
Understand cancer cells at molecular, cellular levels.
Studies of cancer cells illuminate mechanisms of normal cell behavior

3. The Development and Causes of Cancer
Tumor: abnormal proliferation of cells:
Benign tumor
confined to original location, not invade
normal tissue, not spread to distant sites
Malignant tumor
invades surrounding normal tissue, spreads through body via circulation or lymphatics (metastasis)
termed cancers.
Fig. 18.1Pancreatic cancer (purple stained nuclei) in normal

4. Development and Causes of Cancer
Most cancers three main groups:
Carcinomas: malignancies of epithelial cells (about 90% of human cancers).
Sarcomas (rare in humans): solid tumors of connective tissue, (muscle, bone, cartilage, fibrous tissue)
Leukemias and lymphomas: blood-forming cells and cells of immune system, respectively.
Tumors are further classified according to tissue of origin and type of cell involved.

5. Note most common cancers; most lethal
Cell5e-Table jpg

6. Most cancers develop late in life
Fig Tumor clonality
Tumor clonality
Fundamental feature of cancer
Tumors develop from single cell that proliferates abnormally (evidence from X-inactivation pattern)
Most cancers develop late in life
Cancer is multistep process:
Cells gradually become malignant
through progressive alterations
Multiple abnormalities accumulate
Selection for growth advantage
Ex: colon cancer increases with age.
Cell5e-Fig jpg
Fig clonality
Fig age and colon cancer

7. Fig 18.4 Stages of tumor development
Tumor initiation: genetic alteration → abnormal proliferation of single cell, → population of clonal tumor cells.
Tumor progression: additional mutations occur within cells of population
Growth advantage,
Survival
Invasion
Metastasis
Single-cell origin from analysis of X chromosome inactivation (female cancers).
Fig. 18.4

8. Fig 18.5 Development of colon carcinomas
EX. Colon cancer
Proliferation of epithelial cells → small benign neoplasm
(adenoma or polyp).
Clonal selection →
Growth of adenomas
of increasing size,
proliferative potential
Becomes carcinoma
metastasis
Cell5e-Fig jpg
Fig. 18.5

9. The Development and Causes of Cancer
Carcinogens - substances that cause cancer.
Radiation, chemicals, viruses can damage DNA, induce mutations.
Solar UV radiation is major cause of skin cancer.
Aflatoxin produced by some molds that contaminate peanuts and other grains
Viruses can cause cancer
People can inherit cancer-susceptibility genes (oncogenes, damaged tumor suppressor genes)]
Tobacco smoke carcinogenic chemicals:
benzo(a)pyrene,
dimethylnitrosamine,
nickel compounds
(nearly 1/3 cancer deaths).
Fig. 18.6

10. The Development and Causes of Cancer
Tumor promoters stimulate cell proliferation:.
Hormones, particularly estrogens, are tumor promoters in some human cancers.
exposure to excess estrogen increases likelihood woman will develop uterine cancer.
Some viruses cause cancer:
liver (HBV), cervical carcinoma (HPV)
Bacterium Heliobacter pylori causes stomach cancer.
* Studies of tumor viruses identify molecular events in development of cancers.

11. Properties of cancer cells
Characteristic properties of Cancer cells:
1. lack density-dependent inhibition of proliferation
2. reduced requirements for growth factors
3. less regulated cell-cell, cell-matrix (less adhesive)
4. not sensitive to contact inhibition
5,6. secrete proteases for invasion, growth factors for angiogenesis
7. don’t differentiate normally, stay undifferentiated
8, 9. not undergo apoptosis; even after DNA damage
10. unlimited DNA replication; (over) express telomerase

12. Cancer cells have lost normal control – 10 properties
Cancer cells lack Density-dependent inhibition - Continue growing to high densities
Normal cells proliferate to finite cell density, (availability of growth factors).
cease proliferating, arrest in G0
Cell5e-Fig jpg
Fig. 18.7

13. The Development and Causes of Cancer
2. Cancer cells reduced requirements for growth factors contributes to unregulated proliferation.
Can stimulate own proliferation (autocrine stimulation).
Fig. 18.8

14. The Development and Causes of Cancer
3. Cancer cells are less regulated by cell-cell, cell- matrix interactions:
Reduced expression of adhesion molecules contributes to invasion, metastasis
Loss of E-cadherin (main adhesion molecule), aids development of carcinomas (epithelial cancers).
4. Cancer cells lack
Contact inhibition
Normal fibroblasts migrate until
contact neighbor cell, stop, adhere.
Tumor cells move after contact
with neighbor cells, migrate over
adjacent cells, grow in disordered,
multilayered patterns.
Fig. 18.9

15. The Development and Causes of Cancer
5,6. Cancer cells secrete:
Proteases to digest extracellular matrix
Growth factors for angiogenesis
Permits invasion of normal tissues
Digest collagen allows penetration of basal laminae, invasion of connective tissue.
New blood vessels needed after tumor about 106 cells; penetrate capillaries

16. The Development and Causes of Cancer
7. Cancer cells don’t differentiate normally.
Most fully differentiated cells cease cell division.
Ex. Leukemias:
Blood cells from hematopoietic
stem cells in bone marrow.
Leukemic cells don’t undergo
terminal differentiation;
arrest at early stages,
retain capacity for proliferation
Fig

17. The Development and Causes of Cancer
8.9. Cancer cells fail to undergo programmed cell death or apoptosis:
Longer life span than normal; not require growth factors
Lack of apoptosis after DNA damage increases resistance of cancer cells to irradiation, chemotherapeutic drugs, (which damage DNA)
10. Cancer cells overexpress telomerase:
Capacity for unlimited DNA replication
Telomerase required to maintain
ends of chromosomes after replication
Fig telomerase

18. The Development and Causes of Cancer
Ex. Assay for cell transformation in vitro:
Study of tumor induction by radiation, chemicals, or viruses
Detect conversion of normal cells to tumor cells in culture (altered growth properties)
Focus assay (1958) recognizes group of transformed cells morphologically distinct “focus” versus normal cells on dish.
Fig Focus: RSV and chicken fibroblasts

19. 18.2 Tumor viruses Tumor Viruses
directly cause cancer in humans or animals
Critical role in research - models for cellular, molecular study
Small genomes allowed identification of viral genes responsible for cancer induction.

20. Hepatitis B and C viruses
Tumor Viruses
Hepatitis B and C viruses
Principal causes of liver cancer.
Viruses infect liver cells, long-term chronic infections, associated with high risk of liver cancer.
HBV is DNA virus HCV is RNA virus

21. Simian virus 40 (SV40, monkey) (and polyomavirus, mice)
Tumor Viruses
Simian virus 40 (SV40, monkey) (and polyomavirus, mice)
not cause human cancer, important model
small genome sizes
Replicates in permissive host
Transforms non-permissive host
(inactivates Rb)
Early region encodes proteins
(small and large T antigens)
Proteins stimulate host cell
gene expression,
DNA synthesis.
Figs ,13

22. Papillomaviruses are small DNA viruses.
Tumor Viruses
Papillomaviruses are small DNA viruses.
About 100 different types infect epithelial cells.
Some cause benign tumors (warts); others cause malignant carcinomas, particularly cervical cancer
Transformation by expression of early genes, E6,E7:
E7 binds Rb; E6 stimulates degradation of p53.
Fig

23. Tumor Viruses
Adenoviruses large family of DNA viruses not associated with human cancer, important models.
Adenoviruses lytic in cells of their natural hosts, can induce transformation in nonpermissive cells.
Adenoviruses potential gene therapy vector

24. Tumor Viruses
Herpesviruses among the most complex viruses, enveloped DNA genomes 100 to 200 kb:
HSV-1, HSV-2 cause
cold sores, genital sores
Varicella zoster virus (VZV)
causes chicken pox, shingles
Kaposi’s sarcoma-associated herpesvirus causes Kaposi’s sarcoma (common with AIDS)
Epstein-Barr virus cause mononucleosis (transient) and Burkitt’s lymphoma

25. Tumor Viruses
Retroviruses (RNA genome, DNA intermediate) cause cancer in animals, including humans.
HTLV-I causes adult T-cell leukemia
Human immunodeficiency virus (HIV) causes AIDS.
HIV not cause cancer directly, but infects & destroys T cells; AIDS patients malignancies (lymphomas, Kaposi’s sarcoma)
Most retroviruses only 3 genes (gag, pol, and env):
for virus replication, not transformation; rarely induce tumors
Other retroviruses have
specific extra genes
induce cell transformation,
oncogenes (carcinogens
Fig

26. Fig 18.16 Cell transformation by RSV and ALV
18.3 Oncogenes - genes that transform cells:
Rous sarcoma virus (RSV)
Prototype highly oncogenic retrovirus
1st oncogene identified by
comparison of RSV to ALV
(avian leukosis virus does
not induce tumors)
** Cellular oncogenes are
involved in development
of non-virus-induced cancers.
Cell5e-Fig jpg
Fig

27. RSV mutants revealed gene responsible for tumors
Fig The RSV genome
RSV mutants revealed gene responsible for tumors
RSV causes sarcomas: oncogene is src. (not in ALV)
Src was 1st protein-tyrosine kinase identified.
* other oncogenic retroviruses have key proteins of
cell signaling path: ras, raf, myc, erbB, fos, jun,
Cell5e-Fig jpg
Fig

28. Cell5e-Table jpg

29. Cell5e-Table jpg

30. Fig 18.18 Isolation of Abelson leukemia virus
Viral oncogenes are derived from genes of host cell:
Key expt: isolation of oncogenic retrovirus Abelson leukemia virus from mice injected with a nontransforming MuLV virus.
One mouse developed lymphoma from which a new, highly oncogenic virus was isolated:
Virus contained an oncogene (abl)
Abl is related to normal cell gene
Normal gene called proto-oncogene
Cell5e-Fig jpg
Fig

31. *18.3 Proto-oncogenes: Oncogenes
Normal-cell genes from which oncogenes originated
Often proteins of signal transduction pathways that control cell proliferation (e.g., src, ras, and raf )
Retroviral oncogenes differ from proto-oncogenes:
transcribed under control of viral promoter, enhancer
expressed at much higher levels, or in inappropriate cells.
point mutations lead to unregulated activity (ex. Ras)
can differ in structure and function from normal proteins:
Fig Raf oncogene: loss of regulatory domain makes oncogene protein unregulated

32. Detection of human tumor oncogene by gene transfer
Evidence cellular oncogenes in human tumors
(gene transfer experiments 1981)
DNA from human bladder
carcinoma induced transformation
of mouse cells in culture,
indicating tumor had oncogene
Many other examples later:
ras, raf, c-myc
erbB, cdk4, PDGFR
Cell5e-Fig jpg
Fig

33. Cell5e-Table jpg

34. Cell5e-Table jpg

35. Fig 18.21 Point mutations in ras oncogenes
First human oncogene: homolog of rasH oncogene of Harvey sarcoma virus.
3 members of ras gene family (rasH, rasK, rasN) are oncogenes most frequent in human tumors.
ras oncogenes have point mutations critical sites:
Mutations maintain Ras proteins constitutively in active GTP-bound conformation
Cell5e-Fig jpg
Fig mutation in RasH of bladder cancers

36. *Cancer cells have abnormal chromosomes:
Oncogenes
*Cancer cells have abnormal chromosomes:
translocations, duplications, deletions.
Creates oncogenes from proto-oncogenes:
altered promoters, gene fusions, amplification of normal gene
Ex Burkitt’s lymphomas:
translocations involve genes
encoding immunoglobulins.
c-myc transcription factor
Expressed abnormally from
immunoglobulin promoter
Tumors of B cells
Fig

37. Translocations rearrange coding sequences → abnormal gene products.
Oncogenes
Translocations rearrange coding sequences → abnormal gene products.
Ex: Translocation of c-abl proto-oncogene causes chronic myeloid leukemia (CML);
fusion protein has constitutive Abl tyr-kinase activity
Fig Bcr-Abl

38. *Proto-oncogene proteins have normal roles in
Oncogenes
*Proto-oncogene proteins have normal roles in
growth factor-stimulated
signal transduction pathways.
Ex. ERK pathway oncogenes:
polypeptide growth factors,
growth factor receptors (ErbB),
intracellular signaling proteins
(Ras, Raf, MEK, ERK),
transcription factors (fos),
transcription targets (cyclin D1).
Fig ERK path
See also Fig

39. Mechanism of Tel/PDGFR oncogene activation
Many oncogenes encode growth factor receptors, mostly protein-tyrosine kinases.
Ex. Receptor for PDGF converted to oncogene by chromosome translocation, replacement of amino terminus by transcription factor Tel.
Fusion protein
Tel sequences
dimerize in absence
of PDGF, →
activate oncogene
tyr protein kinase.
Cell5e-Fig jpg
Fig

40. Mutant fos or jun always activate Cyclin D1 proto-oncogene,
Oncogenes
Oncogenes can encode transcription factors normally induced by growth factors
EX. Transcription of fos proto-oncogene is induced by phosphorylation of Elk-1 by ERK (Fig ).
Fos and Jun dimerize to form AP-1 transcription factor, activates transcription of cyclin D1
Mutant fos or jun always activate
Cyclin D1 proto-oncogene,
can become oncogene
(CCND1) by chromosome
translocation or
gene amplification
Fig

41. Oncogenes
Oncogenic activity of transcription factor can result from inhibition of differentiation.
Ex. Mutated form of retinoic acid receptor (PML/RARa) oncoprotein in acute promyelocytic leukemia (APL)
Mutated receptors interfere with
action of normal homologs,
block cell differentiation,
maintain leukemic state.
Treatment with retinoic acid
induces differentiation,
blocks cell proliferation.
Fig

42. Tumor Suppressor Genes
*18.4 Tumor suppressor genes normally act to inhibit cell proliferation and tumor development.
In many tumors, both genes are lost or inactivated, contributes to abnormal proliferation of tumor cells
Tumor suppression first noticed during somatic cell hybridization experiments in 1969; hybrids did not cause tumor, suggesting genes in normal cell suppressed tumors.
Fig

43. Fig 18.31 Inheritance of retinoblastoma
First tumor suppressor gene found was retinoblastoma, inherited childhood eye tumor.
About 50% of children of affected parent develop retinoblastoma: inherited as dominant autosomal susceptibility to tumor development:
Need somatic mutation of other copy
photo
Cell5e-Fig jpg
Fig ; retinoblastoma;
Purple, affected people

44. Tumor Suppressor Genes
Retinoblastoma requires loss of both functional copies of tumor susceptibility gene (Rb gene);
People inherit one mutated gene, later other mutates
Fig

45. Mutations of Rb during retinoblastoma development
Noninherited retinoblastoma is very rare:
needs two independent somatic mutations in cell.
Cell5e-Fig jpg
Fig

46. Tumor Suppressor Genes
Rb is tumor suppressor:
Deletions of chromosome 13q14 in some retinoblastomas suggested loss (rather than activation) of Rb gene led to tumor development
Mutations of Rb contribute to many human cancers
Oncogene proteins of some DNA tumor viruses, including SV40, adenoviruses, and human papillomaviruses, bind to Rb and inhibit it. .
Fig ,34

47. Mutations ruining tumor suppressor genes:
common molecular alterations in human tumors
Cell5e-Table jpg

48. Tumor Suppressor Genes
Mutations ruining tumor suppressor genes are common molecular alterations in tumors
p53 - second tumor repressor gene identified.
inactivated in many cancers, leukemias, lymphomas, sarcomas, brain tumors, carcinomas.
Mutations of p53 in about 50% of all cancers.
Proteins encoded by most tumor suppressor genes inhibit cell proliferation or survival.
Tumor suppressor proteins inhibit regulatory pathways stimulated by products of oncogenes.

49. Tumor Suppressor Genes
Tumor suppressor proteins inhibit pathways stimulated by products of oncogenes.
Ex. Products of Rb and INK4 (p16)
tumor suppressor genes regulate
cell cycle at restriction point in G1
Point affected by
cyclin D1/ Cdk4
(which can be oncogenes).
Fig

50. Tumor Suppressor Genes
Ex. p53 gene product regulates both cell cycle progression and apoptosis (programmed cell death)
DNA damage induces p53,
activates cell cycle inhibitory gene p21,
transcription of proapoptotic genes
Cells lacking p53 do not cycle arrest,
not do apoptosis after
DNA damaging agents
p53 mutated in many cancers
(Fig p21 inhibits Cdk2/cycE)
Fig

51. Tumor Suppressor Genes
Ex. BRCA1 and BRCA2 genes
(responsible for some inherited breast and ovarian cancers) function as stability genes that maintain integrity of genome:
Checkpoints in cell cycle progression,
Repair of DNA double-strand breaks
Mutations inactivating these genes leads to a high frequency of mutations in oncogenes or tumor suppressor genes.

52. Tumor Suppressor Genes
MicroRNAs (miRNAs) also regulate gene expression
post-transcriptionally, inhibit translation and/or induce mRNA degradation,
contribute to regulation of about 1/3 of protein-coding genes.
Expression of miRNAs is lower in tumors, suggesting they are tumor suppressors.
Example: let-7 targets oncogene c-myc
[Other miRNAs may be oncogenes]
Fig

53. Tumor Suppressor Genes
Development of cancer is multistep process:
Accumulated damage in multiple genes → increased proliferation, survival, invasiveness, metastatic potential.
Large-scale genome sequencing detects frequency mutations
100 colorectal cancers: each tumor ~ 15 mutations in genes thought to be involved in cancer development:
Oncogenes rasK and PI3K ; Tumor suppressor genes APC and p53.
Breast cancers ~ 14 mutations per tumor, p53 tumor suppressor and PI3K oncogene
Fig

54. Molecular Approaches to Cancer Treatment
Molecular defects being deciphered-> new approaches to prevention and treatment
1. prevent development of cancer
2. detect early premalignant, before metastasis
3. identify susceptible individuals (gene tests)
4. molecular diagnosis of oncogenes, tumor suppressor genes in patient
5. drugs specifically targeted to mutant proteins

55. Molecular Approaches to Cancer Treatment
Early detection of cancer:
Cured by localized treatment, before metastasis.
Ex. Early stages of colon cancer (adenomas) usually curable by minor surgical procedures.
Fig

56. Molecular Approaches to Cancer Treatment
Identify individuals with inherited susceptibilities to cancer development, diagnose tumors.
Mutations in tumor suppressor genes (p53, Rb), oncogenes (myc and cdk4), stability genes (BRCA1 and BRCA2) detected with genetic testing.
Molecular analysis of oncogenes, tumor suppressor genes used in diagnosis of tumor (Bcr-Abl, ErbB-2)
Molecular markers monitor course of disease during treatment (loss of PML/RAR with treatment).

57. Molecular Approaches to Cancer Treatment
Treat with drugs that inhibit angiogenesis:
Blocks proliferation of endothelial cells, less toxic to normal cells.
Most cancer drugs damage DNA or inhibit DNA replication, are toxic to normal cells
Especially cells continually replaced by division of stem cells (hematopoietic cells, epithelial cells of gastrointestinal tract, hair follicle cells)

58. New drugs targeted specifically against oncogenes
recall proto-oncogenes important roles in normal cells.
Acute promyelocytic leukemia (APL):.
Gene that encodes retinoic acid receptor (RARa) fused with PML to form PML/RARa oncogene.
Treatment with retinoic acid differentiates cells, remission of leukemia in most patients.
Breast cancer:
Herceptin - monoclonal antibody against ErbB-2 oncogene protein, which is over-expressed in 25–30% of breast cancers (amplification of erbB-2 gene).

59. Molecular Approaches to Cancer Treatment
Small molecule inhibitors of oncogene proteins, protein kinases:
Imatinib or Gleevec, specific inhibitor of Bcr/Abl protein kinase, blocks proliferation of chronic myeloid leukemia cells (CML).
Imatinib inhibits PDGF receptor and Kit protein-tyrosine kinases:
Kit oncogene in most gastrointestinal stromal tumors.
Imatinib active against tumors with PDGF receptor activated as oncogene
Catalytic domain abl plus imatinib

60. Molecular Approaches to Cancer Treatment
gefitinib and erlotinib, (small molecule inhibitors of EGF receptor), active against some lung cancers.
Responsive lung cancers had mutations for constitutive activation EGF receptor tyrosine kinase.
Fig