Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658

Henrietta Lacks

Cancer: Detection and Treatment

Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658

Henrietta Lacks: the source of HeLa cells

Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658

Henrietta Lacks: the source of HeLa cells“During the 1970s and 1980s, as many as one in three cell lines deposited in cell culture repositories were imposters, one cell line overtaking or masquerading as another. The most notorious culprit was a cervical carcinoma line, HeLa, established by George Gey at the Johns Hopkins Medical School in 1951”

Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658

Henrietta Lacks: the source of HeLa cells

An estimated $10 million of research was discredited.

Cancer:

• is the loss of control over cell division.

• Tumors are normal cells that are dividing inappropriately.

– They stop performing their “normal” function, and are dividing repeatedly.

A cell becomes cancerous when there are incorrect positive AND negative

signals.

Multiple mutations are required for cancer to occur

Fig22.17

Fig. 3 TRENDS in Ecology and Evolution Vol 21 pg 47

Balance between Longevity and Health

Causes of mutations:

• Replication errors– Exacerbated by poor DNA repair– Limited by telomere length

• Other biological agents– Viruses– Transposons

• Environmental factors– Ultraviolet light– Mutagenic chemicals

• smoking, industrial waste, natural toxins

Environment plays a large role in the chance of contracting cancer…

The multiethnic cohort study: exploring genes, lifestyle and cancer risk. L Kolonel, D Altshuler, B Henderson (July 2004) Nature Reviews Cancer 4, 519-527 Fig 1

Chernobyl fireApril 26, 1986

http://video.google.com/videoplay?docid=1108163171078608385

The Chernobyl accident was by far the largest unintentional release of radioactive material into the environment and caused widespread contamination in Europe.

After the accident on April 26, 1986, ~116,000 people were evacuated, most from a zone of 30-km radius. That included about 45,000 people from the town of Pripyat.

The most heavily exposed emergency workers received doses that were sufficiently high to kill them in the weeks and months after the accident.

Chernobyl April 26, 1986

Cancer consequences of the Chernobyl accident: 20 years on J. Radiological Protection26 (2006) 127–140Elisabeth Cardis et al.

The rapidity of increased childhood thyroid cancer in the heavily contaminated areas of Belarus, Ukraine and Russia was surprising.

4 years

The issue of the effects on health of the Chernobyl accident has become part of the political debate over the future role of nuclear energy, which has inevitably led to dispute over the level of effects either observed or anticipated.

After the accident on April 26, 1986, ~116,000 people were evacuated, most from a zone of 30-km radius. That included about 45,000 people from the town of Pripyat.

For more pictures: http://www.nikongear.com/Chernobyl/Chernobyl_1.htm

Source: US Mortality Public Use Data Tape 2000, National Center for Health Statistics, Centers for Disease Control and Prevention, 2002.

1. Heart Diseases 710,760 29.6

2. Cancer 553,091 23.0

3. Cerebrovascular diseases 167,661 7.0

4. Chronic lower respiratory diseases 122,009 5.1

5. Accidents (Unintentional injuries) 97,900 4.1

6. Diabetes mellitus 69,301 2.9

7. Influenza and Pneumonia 65,313 2.7

8. Alzheimer’s disease 49,558 2.1

1. Nephritis 37,251 1.5

10. Septicemia 31,224 1.3

Rank Cause of Death # of deaths

% of all deaths

US Mortality, 2000

Change in the US Death Rates* by Cause, 1950 & 2000

* Age-adjusted to the 2000 US standard population.Source: US Mortality Volume 1950, National Vital Statistics Report, 2002, Vol. 50, No. 15.

586.8

180.5

48.160.923.7

200.9193.7

258.2

0

100

200

300

400

500

600

HeartDiseases

CerebrovascularDiseases

Pneumonia/Influenza

Cancer

1950

2000

Rate Per 100,000

2003 Estimated US Cancer Cases*

*Excludes basal and squamous cell skin cancers and in situ carcinomas except urinary bladder.Source: American Cancer Society, 2003.

Men675,300

Women658,800

32% Breast

12% Lung & bronchus

11% Colon & rectum

6% Uterine corpus

4% Ovary

4% Non-Hodgkin lymphoma

3% Melanomaof skin

3% Thyroid

2% Pancreas

2% Urinary bladder

20% All Other Sites

Prostate 33%

Lung & bronchus 14%

Colon & rectum 11%

Urinary bladder 6%

Melanoma of skin 4%

Non-Hodgkin lymphoma 4%

Kidney 3%

Oral Cavity 3%

Leukemia 3%

Pancreas 2%

All Other Sites 17%

2003 Estimated US Cancer Deaths*

ONS=Other nervous system.*Excludes basal and squamous cell skin cancers and in situ carcinomas except urinary bladder.Source: American Cancer Society, 2003.

Men285,900

Women270,600

25% Lung & bronchus

15% Breast

11% Colon & rectum

6% Pancreas

5% Ovary

4% Non-Hodgkin lymphoma

4% Leukemia

3% Uterine corpus

2% Brain/ONS

2% Multiple myeloma

23% All other sites

Lung & bronchus 31%

Prostate 10%

Colon & rectum 10%

Pancreas 5%

Non-Hodgkin 4%lymphoma

Leukemia 4%

Esophagus 4%

Liver/intrahepatic 3%bile duct

Urinary bladder 3%

Kidney 3%

All other sites 22%

Cancer Death Rates*, for Men, US, 1930-1999

*Age-adjusted to the 2000 US standard population.Source: US Mortality Public Use Data Tapes 1960-1999, US Mortality Volumes 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2002.

0

20

40

60

80

100

1930 1935 1940 1945 1950 1955 1960 1965 1970 1975 1980 1985 1990 1995

Lung

Colon and rectum

Prostate

Pancreas

Stomach

Liver

Rate Per 100,000

Leukemia

Cancer Death Rates*, for Women, US, 1930-1999

*Age-adjusted to the 2000 US standard population.Source: US Mortality Public Use Data Tapes 1960-1999, US Mortality Volumes 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2002.

0

20

40

60

80

100

1930 1935 1940 1945 1950 1955 1960 1965 1970 1975 1980 1985 1990 1995

Lung

Colon and rectum

Uterus

Stomach

Breast

Ovary

Pancreas

Rate Per 100,000

Tobacco Use in the US, 1900-1999

0

500

1000

1500

2000

2500

3000

3500

4000

4500

5000

190019051910191519201925193019351940194519501955196019651970197519801985199019952000Year

Per Capita Cigarette Consumption

0

10

20

30

40

50

60

70

80

90

100

Age-Adjusted Lung Cancer Death

Rates*

*Age-adjusted to 2000 US standard population.

Source: Death rates: US Mortality Public Use Tapes, 1960-1999, US Mortality Volumes, 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2001. Cigarette consumption: Us Department of Agriculture, 1900-1999.

Per capita cigarette consumption

Male lung cancer death rate

Female lung cancer death rate

Tobacco Use in the US, 1900-1999

0

500

1000

1500

2000

2500

3000

3500

4000

4500

5000

190019051910191519201925193019351940194519501955196019651970197519801985199019952000Year

Per Capita Cigarette Consumption

0

10

20

30

40

50

60

70

80

90

100

Age-Adjusted Lung Cancer Death

Rates*

*Age-adjusted to 2000 US standard population.

Source: Death rates: US Mortality Public Use Tapes, 1960-1999, US Mortality Volumes, 1930-1959, National Center for Health Statistics, Centers for Disease Control and Prevention, 2001. Cigarette consumption: Us Department of Agriculture, 1900-1999.

Per capita cigarette consumption

Male lung cancer death rate

Female lung cancer death rate

Treating cancer:• Avoid it

– Avoid mutagens– DNA repair gets less efficient as we age

T-cells recognize and eliminate abnormal cells; such as cells with many mutations

Our immune system protects us from cancer

P53 is activated by DNA damage

Fig 22.15

p53 can induce apoptosis via two pathways: Nuclear and/or Mitochondrial

Treating cancer:• Avoid it

– Avoid mutagens– DNA repair gets less efficient as we age

• Surgery– Must remove all cancer cells– Non-invasive

Treating cancer:• Avoid it

– Avoid mutagens– DNA repair gets less efficient as we age

• Surgery– Must remove all cancer cells– Non-invasive

• Radiation– Directed at tumor; causes DNA damage

-> cellular self-destruction– Mutagenic, side effects

Treating cancer:• Avoid it

– Avoid mutagens– DNA repair gets less efficient as we age

• Surgery– Must remove all cancer cells– Non-invasive

• Radiation– Directed at tumor – Mutagenic, side effects

• Chemotherapy– Toxins directed at rapidly dividing cells– Mutagenic, many side effects

Chemotherapy

a rapidly dividing cell

Toxin

X X

Normal Multi-Drug Resistance protein

MDR

MD

RMDR

MD

Rtoxin/hormone/etc

toxin/hormone/etc

toxin/h

ormon

e/etctoxi

n/h

orm

one/

etc

Some cancers over-express MDR

Toxin

MDR

MD

R

MDRMDRMDR

MD

RMDRMDRMDRMDR

MD

RM

DR

toxin toxin

toxin toxin toxin toxin

toxintoxinto

xin

toxi

n

toxin toxin

I’m a cancer cell with over-expressing MDR. I laugh at your toxins.

The Epigenetic Progenitor Origin of Human Cancer (2007) A P Feinberg, R Ohlsson, S Henikoff Nature Reviews Genetics 7: 21-31

Mutations continue after cancer develops

OO

OOOO

OOO

OOCancer cell with mutation causing MDR over-production

Evolution: changes in DNA as information transmitted

OO

OOOO

OOO

OO

OO

OOOO

OOO

OO

Applychemo-therapy

XXX

XX X XX

XX

Kills most cells.Except if some have mutation that allow them to be resistant.

Evolution: changes in DNA as information transmitted

Cancer cell with mutation causing MDR over-production

OO

OOOO

OOO

OO

OO

OOOO

OOO

OO

O

XXX

XX X XX

XX

Kills most cells.Except if some have mutation that allow them to be resistant.

Continues to replicate

Evolution: changes in DNA as information transmitted

Applychemo-therapy

Cancer cell with mutation causing MDR over-production

OO

OOOO

OOO

OO

OO

OOOO

OOO

OO

OO

OOOO

OOO

OO

O

XXX

XX X XX

XX

Kills most cells.Except if some have mutation that allow them to be resistant.

Continues to replicate

Tumor with cells expressing MDR

Evolution: changes in DNA as information transmitted

Applychemo-therapy

Cancer cell with mutation causing MDR over-production

Some cancers over-express MDR

Toxin

MDR

MD

R

MDRMDRMDR

MD

RMDRMDRMDRMDR

MD

RM

DR

toxin toxin

toxin toxin toxin toxin

toxintoxinto

xin

toxi

n

toxin toxin

I’m a cancer cell with over-expressing MDR. I laugh at your toxins.

Detecting Cancer or Types of Cancer

Cancer Cells

Normal Cells

A Microarray is a chip with DNA sequences (genes) bound to the surface at known locations.

It can be used to track or monitor expression of many genes.

Tracking changes in gene expression using a Microarray

Making cDNA from RNA

Tracking changes in gene expression using a Microarray

Tracking changes in gene expression using a Microarray

Use of microarray to estimate genes likely present in malignant cancers

Patients cancer free for 5+ years

Patients cancer spread in 5 years

Use of microarray to estimate genes likely present in malignant cancers

similar to Fig 22.19

different genes

Fig 22.19 Microarrays can be used to get information about types of cancers

Young (>55) Breast cancer patients

More accurate profiling of tumors results in more accurate choices of treatments. Patients with benign tumors can avoid chemotherapy (adjuvant).

Effect of active smoking on the human bronchial epithelium transcriptome (2007)R Chari, K M Lonergan, R T Ng, C MacAulay, W L Lam, and S LamBMC Genomics, 8:297

Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297CS=current smoker, FS=former smoker, NS=never smoked

Table 1: Subject Demographics

Overlapping and unique genes expression

Fig 1B

Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297

Effect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297

Table 3: Reversible gene expression upon smoking cessation related to mucus secretion (genes in bold have not been previously associated with smoking)

CABYR ENTPD8 TFF3

Fig 4AEffect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297

Some changes in gene expression induced by smoking are reversible

MUC5AC GSK3B

Fig 4BEffect of active smoking on the human bronchial epithelium transcriptome (2007) R Chari et el. BMC Genomics, 8:297

Smoking can induce irreversible changes in gene expression

Treating cancer:• Avoid it

– Avoid mutagens– DNA repair gets less efficient as we age

• Surgery– Must remove all cancer cells– Non-invasive

• Radiation– Directed at tumor – Mutagenic, side effects

• Chemotherapy– Toxins directed at rapidly dividing cells– Mutagenic, many side effects

Cell Culture Forensics. S. O’brien PNAS July 3, 2001 vol. 98 no. 14 7656-7658

Henrietta Lacks

Cancer: Detection and Treatment