How to Understand and Treat Cancer with Molecular Markers

How to Treat Cancer with Molecular MarkersHow to Treat Cancer with Molecular MarkersHow to Treat Cancer with Molecular MarkersHow to Treat Cancer with Molecular Markers

Presented by Professor Serge JurasunasPresented by Professor Serge JurasunasPresented by Professor Serge JurasunasPresented by Professor Serge Jurasunas

Lisbon, PortugalLisbon, PortugalLisbon, PortugalLisbon, Portugal

www.sergejurasunas.comwww.sergejurasunas.comwww.sergejurasunas.comwww.sergejurasunas.com

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1st of a Series on Naturopathic Oncology

How to Understand and Treat

Cancer with Molecular Markers

Presented by:

Serge Jurasunas N.D., M.D. (HOM)

www.sergejurasunas.com

Member of the Society of Integrative Oncology

American Naturopathic Medical Association2

Lecture Overview

→ What is cancer?

→ What metastasis means?

→ The need for new diagnostics

→ P53 Tumor supressor gene

→ Novel approach to cancer with dietary agents

→ Clinical cases

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1938 American Society for the

Control of Cancer poster

What is Cancer?

What causes cancer?

Why cancer cannot be

cured?

What is the link between

environment, food and

cancer?

How cancer can be

diagnosed?

Can cancer be prevented?

What is the link between

genes and cancer?

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Interactions Between Genes and the Environment

Susceptibility to cancer

En

vir

on

me

nt

insu

lts

Affected

Unaffected

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Where are We Today with Cancer?

Despite continuing improvement in conventional

diagnosis, local management and some progress in

chemotherapy...

There has been no significant increase in survival

rate over the past 35 years.

Resistance to chemotherapeutic drugs is

considered the greatest obstacle to the successful

management of cancer patients.

Cancer recurrence is still too high.

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First Question: What Cancer Means?Carcinoma derived from the Greek Karkinos – Crab: Hyppocrates (460-375 B.C.)

Cancer defines a population of

abnormal, damaged cells that have

lost their normal control and

differentiation, divide unchecked,

evades apoptosis, accumulates

mutations and keeps proliferating.

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Second Question: What Does Metastasis Mean?

Metastasis is the process by

which a tumor cell leaves the

Primary site through a lack of

cellular attachment and

addhesion.

Breaks the basal membrane

degrade the E.C.M. and

penetrate into the blood

circulation,lymphatic

circulation.

Travels to distant sites, survives

immune defenses and

establishes a secondary tumor.

Metastases are responsible for 90% of cancer mortality.

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These are anti-apoptotic cells, stem

cells resistant to destruction.

Metastases may enter into the G.O.

dormancy phase of the cell cycle and

do not divide or proliferate, keeping

silence during the ensuing months or

years.

Under stimuli, infection, metastases

may reactivate and re-enter in the cell

cycle and proliferate.

Metastases

Malgarzata Bany et al – Dormancy in breast cancer: Breast Cancer: targets and Therapy: 2012:4 – 183-1410

Conventional Diagnostics

1. Inefficient in cancer prevention.

2. Many cancer and pre-malignant forms

go undetected, asymptomatic for

many years .

3. Inefficient to monitor cancer cell

resistance to chemo/radiation.

4. Does not select the best adapted

treatment.

5. Inefficient to predict disease

recurrence.

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Molecular Medicine and Cancer Biology

Molecular medicine is

important in order to detect

pre-malignant conditions

cancer initiation and/or in

earlier stage before it starts

to proliferate and invade the

body.

P53

mutation

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The New Solution to Cancer

Activate the tumor supressor genes own

apoptotic body to destroy cancer cells

through a natural programmed

mechanism called apoptosis.

Apoptosis induction is recognized as the

most potent defense in our body against:

Cancer initiation

Tumor growth

Metastatic spread

Prevention, Diagnosis and Treatment

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Each of Our Cells is Genetically Equiped with a Mechanism

of Cellular Suicide When Cells are Damaged

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In 1993 P53 Appeared on the Cover of the Journal, Science as the

Molecule of the Year

P53 – Tumor Suppressor Gene

the Guardian of Our Genome

Over 60,000 publications

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January 13, 1997

P53 – Regulates the Cell Cycle and all Cell Division

P53 is a tumor suppressor protein that regulates

the cell cycle and is involved in the expression of a

wide variety of genes involved in Apoptosis such

as Bax, Puma, Noxa, BCL-2.

P53 normally functions to arrest the cell cycle and

division when a cell is damaged by external

stimuli (radiation, stress, chemotherapy) or P53

trigger apoptosis.

P53 prevents the proliferation of cells with a

potential for neoplasic transformation.

Under stress stimuli the P53 gene expression is

activated and P53 protein level is increased.

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Cellular Stress or DamageNon Genotoxic stress – Hypoxia – Oxidative stress – Genotoxic stress –

Chemotherapy - Radiation

Under oxidative stress DNA damage

occurs naturally about 10,000 times

per day per cell in the human body.

Damaged cells are either repaired

and continue their cycles or are

destroyed and eliminated through

apoptosis.

The prevention of cancer is

profoundly dependent on the

P53 tumor supressor protein.

Serge Jurasunas 2015

Physiologic stress

Low/middle stress High stressDNA

damage

DNA

damage

P53 protector P53 killer

P53 protein

P21

P53 protein

Puma

Noxa

BAX

Apoptosis

Cell cycle

Arrest

DNA repair

Restart

Genomic stability

Repair

P53

P53P53

BCL-2

Caspases 3-7-9

BAX

BCL2MT

CytC.

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P53 Mutations

1. P53 gene is the most commonly

mutated gene that occurs in

over 50% to 70% of all cancer.

2. P53 mutation appears

necessary for many types of

cancer.

3. P53 mutation leads to induce a

mutant protein.

4. When the P53 apoptosis

pathway is impaired, then

mutant protein gains new

function or oncogenic function.

Eric Wattel et Claude Preudhomme et al – P53 mutations are

associated with resistance to chemotherapy and short survival

in hematologic malignancies – Blood vol.84 nº9 – Nov.11-1994-

3148.3157 20

The Bad News

P53 Mutation is Associated with Every Step of Tumor

Growth, Metastases and Invasion

� Aggressive cancer.

� Angiogenesis.

� Resistance of cancer cells to

chemotherapy/radiation.

� Breast glioma, pancreatic, colon cancer.

� Responsable for the transmission of

cancer in the same family.

1 – Mazars B., Spinardi L., Bencheilch M., Simony Lafontaine, Theillet C.,

- P53 mutations occur in aggressive breast cancer – cancer Res. 1992-52

3918-3923.22

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Causes of P53 Mutation

P53

mutation

Aging

Tobacco

Pesticides

Genotoxic Free

radicals

Virus

Bacteria

Radiation

P53 Mutations as Fingerprints of Environmental Carcinogens

Rengul Cetin – Atalay and Mehnet Ozturk – P53 mutations as fingerprints of environmental carcinogens – Pure APP.chem – Vol.72-nº6-PP995-999-2000

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Novel Approach to Cancer Therapy

Targeting mutant P53 to reactive normal

(wild type) function by using small

molecules.

Increasing apoptosis and self-destruction of

cancer cells – Reactivation of P53 tumor

supressor gene is now considered as a

new anticancer treatment.

Zhen Wang and Yi Sun – Targeting P53 for novel anticancer

approach – Trans L. Oncol. 2010 – Feb. 3 (1) 1-2.

Bykov, V.J. et al (2002) – Restoration of the tumor supressor

funtion to mutant P53 by a low-molecular-weight compound.

Nat.Med.8-282-28825

P53 and Chemotherapy

P53 mutation Chemotherapy Prognosis ?

P53 mutation increase the resistance of cancer cells to

numerous chemotherapy agents that fail to destroy. Cancer

cells can accumulate and invade other tissue.

Eric Wattel, Claude Preudhomme et al… - P53 mutations are associated with resistance to chemotherapy and short

survival in hematologic malignancies – Blood Vol.84 – N.9 – Nov.11 – (1994) 3148 – 3157.26

P53 Mutations Predict Outcome in Advance

65/93 Studies found that P53 is a statistically significant factor of poor prognosis in various cancer.

Studies have found that the presence of a mutation was associated with a poor response to various

chemotherapy or radiotherapy regimens in breast, colorectal, ovarian, stomach and soft tissues

carcinoma.

Texas researchers have found that high levels of P53 mutated protein accumulation are associated

with a significantly increased local recurrence rate in 1,500 breast cancer patients treated with

mastectomy.

P53 – positive patients 21.5%

negative patients 9.3%

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Apoptosis in Tumors treated with Cytotoxic Chemotherapy

is Driven by P53

Growth of Xenograft Tumors

Tumors with intact P53 gene

regressed during the treatment,

whereas the tumors with delected

P53 genes continued to grow.

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We Need to Overcome the Toxicity of Anticancer

agents, radiotherapy, etc.

Toxic toHealthy Cells

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Improving Patient Quality of Life, Less Adverse Effects, and Increasing

Chemotherapy Effectiveness

F. 36 years old – Advanced colorectal cancer

with multiple metastases to liver – follow

our treatment.

Case published in Townsend Letter – April

2009

F. 71 years old – Breast cancer –

chemotherapy with severe toxic adverse

effects – Developed a peripheral neuropathy

– Paralyzed on the first consultation.

The Balance Between Pro-apoptotic and Anti-apoptotic Proteins

Determines the Fate of a Cell

Serge Jurasunas 2013

Cells commit suicide

RadiationOxidative

Stress event

Chemotherapy

Anti-apoptotic signals

BCL2

Survivin

Pro-apoptotic signals

P53 Tumor suppressor

BAX

P21

Apoptosis

DNA

damage

Balance

Survivance

Resistance to Apoptosis

Cells do want to die

... . .

.

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Rational for Utilizing a Combination Cancer TherapyIncrease chemotherapy/radiation effectiveness

Chemotherapeutic agents

Cancer cells

Apoptotic Pathway

P53

Puma

BAX

NOXA

Caspases

Death

Dietary agents - blockers

Anti-apoptosis/survival

BCL2 – BCLx4

NF.KB

Survivin

Survival Resistance

Curcumin

Resveratrol

Pomegranate

Genistan

Indole-3-Carbinol

P53

P21

Reverse P53 mutation

Death

Induce apoptosis

PSJ53 Therapy

Chlorella

Enzyme yeast cells

Antioxidants

compound

Oligonucleotide

Fermented

chlorella

Keith R. Martin – Targeting apoptosis with dietary bio-active agentes – Exp.Biol. Med. 231:117-129-200634

Dietary Agents with Anti-Cancer Properties

Keith R. Martin – Targeting apoptosis with dietary bioactive agents: Nutrition and cancer Lab., Dept of Sciences – Pennsylvania State Uni – USA - 200635

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Modulation of Apoptosis by Active Compounds for Cancer Therapy (1)

Curcumin Lung, pancreas, colon, prostate,

stomach, breast.

Increase chemotherapy sensitivity-

synergetic

Gemcitabline, oxilaplatin,

paclitaxel

5 Fu

P53 – P21 – Survivin – NF.KB –

Cox2 – VEGF – BAX – BCL-2

Resveratrol

Apigenin

Ovarian, breast, prostate, liver,

lung, uterine.

Cisplatin, doxorubicin, platinum

compounds, 5 Fu, paclitaxel

Pancreatic cancer

5Fu, Paclitaxel

P53 – BAX – Caspases activator –

P21 – Survivin – BCL-2 – Cyclin 1 –

Cox2

G1-G2 – S-phase arrest

P53 – P21 – BCL-2 – BAX –

Caspases – C-Myc – VEGF –

induce G 2/ M – Cell cycle arrest

Pomegranate

Indole-3 - Carbinol

Prostate, breast, lung, colon, oral,

leukemia

Breast cancer, prostate, pancreatic

Gemcitabine – cisplatin - paclitaxel

BCL-2 – BAX – VEGF – Cox2 – P21

– BAK – NF.KB

BCL2 – BAX – NF.KB

Activate caspases

Organ Site Molecular target

Ellagic acid Neuroblastoma, pancreas, breast,

prostate, colon, intestine, bladder,

oral, liver.

Cisplatin, vinorelbine

P53 – P21 – Cyclin 1 – Cox 2 –

NF.KB

Increase caspase 3

Quercitin

Capsaicin

Colon cancer, breast.

5Fu, Cisplatin

Doxorubicin

BAX – BCL-2 – Caspase - Cyclin

D1 – Gadd 45

BCL-2 – BAX - Survivin – Cyclin D1

NF-KB – CDK1 – CDK2

Increase Cyt C. release Caspase 3

P53 – BCL-2 – P21

VEGF – Cyclin D1

Catechin 5 Fu, Gemcitabine – mitocycin



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Green tea

Polyphenols and EGCG

Lung, oral, stomach, liver,

pancreas, colon, bladder, prostate,

breast.

Tamoxifen – cisplatin –

adriamycin, dacarbazine

P53 – P21 – BAX – BCL-2 – NF.KB

– Cyclin 1 - Cox2 – VEGF _

increase Xiap

Isithiocyanates (ITC’s)

Sulforaphane

(Cruciferous vegetables)

Prostate, breast

Docetaxel

Adriamycin

Cisplatin

BCL-2 – Survivin – NF.KB – Cyclin

1 – P53 – caspase activation

Licopene Prostate, lung, breast, gastric,

liver, pancreas colorectal

Adriamycin

Cisplatin

Cyclin D1 - BCL-2 – BCL-4 AKT –

NF.KB – MMP9



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Effect of Caffeic Acid Phenethyls Ester (Cape) – (Propolis)

63 Year Old Woman – Infiltrating Ductal Carcinoma

A – Cell’s without treatment.

B – Cells treated with 15 UM CAPE for 48

hours - condensation of apoptotic body.

Photo: immunofluorescence microscopy x 20..

Result: Significant inhibitory effects on the

growth and viability of tumor cells in vitro.

Mohamed F: El Refaei and Essam A. Mady – Regulation of Apoptosis, Invasion and angiogenesis of Tumor cells by CAPE – Intech – Open Science –

Chapter 14 - 201339

Curcumin: Targets P53, BAX, BCL2, Inhibits NF.KB, VEGF, etc.

Melanoma Cells – Apoptotic cells (Curcuma 15UM Micromoles) after 48 hours

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Anticancer Properties of Foods

Apoptosis – Angiogenesis – Inflammation – Immune Defense

“Let food your medicine and medicine be your food.”

Hyppocrates 460-370 B.C.41

Comprehensive Cancer Treatment

Prevention or Cancer Therapy

Immune modulation Antioxidant therapy

Hereditary profile

Health status

Detoxification

Food diet and

dietary agents

Apoptose induction

Inflammation

Angiogenesis

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Patients Undergoing Molecular Markers Tests

Method: Elisa Assay

Quantitative Polymer Chain Reactional (P.C.R.)

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P53 Gene Expression P53 Protein Level

18.788 copies/ml of plasma 57.3 µ of normal protein/ml of plasma

Reference range

50 copies

F – 44 years – Non Cancerous Patient – 2009

(Stressed Cells)

Reference range

0.33 units

Comment:

There is a population of stressed cells with damaged DNA as it is indicated by P53 gene

expression and high level of WT protein.

Cancer lesions are unlikely because of the absence of mutated P53 protein .

The cancer defense is functioning.

P53 Assay

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P53 Assay Examples:

Two Cases of Breast Cancer Remission

F.43 years old

Test done 1 year after remission

P53 Gene Expression

1.3x106 copies/ml of plasma(100.000)

Reference range: = 1000 copies

P53 Protein Level

5.2 µ of normal protein/ml of plasmaReference range: 0.33 copies

Self-destruction of abnormal/stressed

cells

F. 48 years old (April 2012)

Test done 3 months after remission

P53 Gene Expression

Not detectable

Reference range: 10-50 copies

P53 Protein Level

0.02 units of mutated protein/ml of

plasmaReference range: 0.10 – 1.00 units

W.T

Not detectableReference range: 0.02 units

The tumor suppressor gene is not active

and produced only a trace of P53

mutation -stressed/transformed cells are

not self-destructed.

Risk of recurrence45

P53 Gene Expression

P53 Protein Level

mutated

3.285 copies/ml of plasma

15.75 µ of abnormal protein/ml of plasma

Reference range:50 copies

Reference range: 0,33 units/ml of plasma

Patient F - 49 Years Old – Ovarian Cancer –

Stage III - Recurrence

Recurrence in 2009

1.7.2009 18.1.2011 (remission)

P53 Gene Expression

1.2 x 108 copies/ml of plasma

P53 Protein Level

normal

125.0 µ of normal protein/ml of plasma

Reference range: Not detectable

Comments:

Cells are resistant to destructive

agents that trigger apoptosis.

Comments:

The treatment reversed the P53 mutant with

oncogenic function. The tumor suppressor

gene is now very active with an increased

level of normal proteins.

Active process of self-destruction of cancer

cells leading to tumor regression.

P53 Protein Level

normal

Not detectable

Reference range: 0,33 units/ml of plasma

Reference range:50 copies

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Patient M. – 44 Years Old – Clinical Story: Multiple Cancer Recurrence – Bladder – Lung – Adrenal

Surgeries – Radiation – Chemotherapy

1st test

18th February 2009

P53 protein level

26.1 u of abnormal

protein/ml of plasma

P53 gene expression

268 copies/ml of plasma

Mutant protein unable to

trigger the self-destruction

of cancer cells escaping from

apoptose

Chemotherapy ineffective.

2nd test

2nd June 2009

P53 protein level

16.8 u of normal protein/ml of

plasma

P53 gene expression


(100.000 copies)

The treatment reverse the

oncogenic dominance of the P53

to a wild type P53 tumor

suppressor.

Activation of the P53 gene and

normal protein.

Self destruction of cancer cells

3rd Test

7th October 2009

P53 protein level

156 u of normal protein/ml of

plasma

P53 gene expression


Very high increasing level of

P53 protein.

The applied treatment

continues to have a profound

destructive effect on the

population of

abnormal/cancer cells

Treatment to Restore Normal Function of mutated P53 with PSJ53 Therapy (1)

(1) Serge Jurasunas N.D. M.D. (Hom), Olga Galkina Taylor Ph.D. – How to target mutante P53 in a case of multiple cancer recurrence – Townsend Letter August/Sept 2010 68-7247

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F. Breast Cancer: Age 51 Years – High Risk of Recurrence - 2012

Date: 5/11/2010

P53 gene expression

169u/ml of plasma

Reference range: (10-50u)

P53 normal protein level

10.99u/ml of plasma

Reference range: (0.33 units)

Date: 10/08/2010

P53 gene expression

Not detectable

P53 protein level

wild: not detectable

mutated: 33.9 units/ml of plasma

Live blood analysis – infected ground

Patient F. with a 4 Year Complete Remission

of Pancreatic Cancer (Townsend Letter – August/Sept 2009 USA)

P53 Test

1st Test – 4.5.2009

P53 Gene Expression

65.000 copies/ml of plasma

Reference range: 1000 copies

P53 Protein Level

4.5 µ of abnormal (mutated)

protein/ml of plasma

P53 Wild Type

Indetectable

Reference range: 0.33 copies

2nd Test – 4.8.2009

P53 Gene Expression



P53 Protein Level

1.5 µ of normal protein/ml of

plasma


We reversed mutant P53 to a

wild type but the actived gene

produced normal P53 protein

only to some extent. Cancer

cells are not fully self-

destroyed

3rd Test – 17.11.2009

P53 Gene Expression



P53 Protein Level

67.4 µ of normal protein/ml of

plasma


Now the P53 gene is still active

and produces high a level of

P53 normal protein leading to

increased self-destruction of

cancer cells.

Tumor regression can be

anticipated49

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THE ANTI-TUMOUR EFFECTS OF THE APPLIED TREATMENT

02/12/12370 05/12/12461 Ref. Range

Ref.Range

P53 gene expression 200 427 10-50 units/ul of plasma

P53 level mutated ND ND ND units/ml of plasma

P53 level wild ND 0,4 0.10-1.00 units/ml of plasma

Bcl-2 gene expression 8000 796 <10 units/ul of plasma

Bax gene expression 167 1543 10-100 units/ul of plasma

Bcl-2/Bax ratio 0.02 1.93

Survivin gene expression 171 900 <10 units/ul of plasma

P21 gene expression 139 738 10-50 units/ul of plasma

Survivin/p21 ratio 0.8 0.8

VegF gene expression 2353 ND 10-100 units/ul of plasma

A Complementary Approach to Breast Cancer: A Case with Multiple Liver

Metastases is Free from Disease Complete Report – Townsend letter Magazine – August/Sept 2014

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Description Intracellular Extracellular Comments

P53 gene expression 1.327 units 43

Anti-tumor activity

P53 level mutated protein ND

P53 level normal 6 units 25.2

Bcl-2 gene expression 487 866 Increasing cancer cells with BCL-2

Bax gene expression 1.431 411

Anti-tumor activity

Bax/Bcl-2 ratio 2.9

Survivin gene expression 1.752 167 Dominance

Risk of recurrence

P21 gene expression 484 952

Survivin/p21 ratio 0.3 Many cancer cells with BCL-2and

Survivin are destroyed over their

accumulation

Patient – M- 57 Years Old – Tumors of the Bladder, Urethra, Prostate (2011)

Surgery - Chemotherapy

Complete Remission

More Information

Modulation of Apoptosis by active dietary compounds

and their synergistic effects with chemotherapy in

cancer treatment.

Articles on cellular respiration and cancer,

mitochondria, P53 and other molecular markers.

Clinical cases.

Integrative cancer treatment.

www.sergejurasunas.com

Email: [email protected]

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Thank You for

Your Attention!

Don’t Let the Cancer Kill You, but rather Kill the Cancer.

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How to Understand and Treat Cancer with Molecular Markers

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Transcript of How to Understand and Treat Cancer with Molecular Markers