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Mission Possible: The Rigors of Cancer Research Teach an Undergraduate About the Discipline Behind the Breakthroughs

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Duke Health News 919-660-1306

Not only would I perform the assassination again if given
the opportunity, I expect to do it the next morning at the same
time. Fatalities are an unfortunate consequence of war, I
remind myself. And the victims of my cold-hearted onslaught?
White blood cells called lymphocytes that are among the body's
immune-system soldiers in the war against cancer.

Putting aside the pipette, I search for yet another virulent
chemical in my artillery--phenol chloroform that will slice
right through the "phospholipid bi-layer" membrane enclosing
the cells, exposing the protected inner environment to a
war-ravaged outside. For, it is not the cells' death that
interest me, but the genetic treasure they cleverly guard
inside--the acclaimed blueprint for all life, DNA.

Once the chemical breaks the cells open, I tease the stringy
strands of genetic material out of the cells they once
controlled. The DNA sits as a slimy glob the texture of mucus
at the bottom of the plastic tube as I suction off the pool of
sorted cell parts that float above. After snagging the gooey
remains with a pipette, I hesitate as it dangles precariously
above a new tube.

It is a moment of realization.

Just four-and-a-half-weeks into my first lab experience, I
am in the middle of a procedure I had only read about in my
biology textbook: harvesting the DNA containing cancer genes,
using enzymes to break it apart in an analytical experiment,
and then "amplifying" the tiny amounts of product DNA using
DNA-copying polymerase chain reaction (PCR), a sort of
biological Xerox machine for genes.

The PCR procedure involves preparing DNA samples in little
vials for insertion into a machine where an enzyme reaction
makes millions of copies. It's a procedure familiar to any
researcher or laboratory technician, but one that represents a
new and profound lab experience to students like me
unaccustomed to bench research.

Stranger in a Strange Land

Although I was a stranger to the lab those first few weeks
of summer, wading my way through a whole new scientific
vocabulary, cancer was not a new word to my lips. My mother had
been diagnosed with low-grade non-Hodgkin's lymphoma more than
five years before. In those years, our personal war with cancer
had led us to three of the world's foremost cancer centers,
including Duke's, for her chemotherapy, radiation, and bone
marrow transplant. And it led me through a crash course
education that I never could have anticipated.

But nothing I ever read or observed gave me the same
educational experience as my months in the lab. I had always
known the cancer patient, never the cancer. That is not to say
that anyone truly understands the full phenomenon of the
disease. In fact, it may take cancer researchers many more
decades of observation and manipulation of malignant cells in
order to grasp their wily ways. But Duke is making it possible
for students like myself to come along for the ride.

Through a grant from the Howard Hughes Medical Institute,
the Duke Comprehensive Cancer Center (DCCC) can place thirty
students, many of them incoming freshman, in the center's labs
each summer. The eight-week program is supplemented with
lectures by leading researchers, tours of the center's shared
resources, and opportunities to observe in the medical center's
various oncology units. Such experiences allow students to
understand the laboratory work in context of its wider
applications.

With millions of women diagnosed with breast cancer each
year, it is easy to see the society-wide importance of the
research I took on. The Lab of Molecular Immunopathology,
headed by principal investigator Laura Hale is a constituent of
the Duke Breast Cancer Program.

One of the lab's current projects aims at understanding
BRCA2, a gene which, when caused to malfunction by mutation,
can lead to breast cancer. Since the identification of the
breast cancer susceptibility genes BRCA1 and BRCA2 in 1995,
discoveries in large part due to the efforts of the medical
center's Andrew Futreal, researchers worldwide have been
working feverishly to describe the genes' roles in causing
breast cancer.

Teamed with graduate student of pathology Kristina Flores, I
had the opportunity to join that world-wide crusade.

In the waning weeks of the program, I participated in the
lab's novel effort to test functions of the protein coded for
by the BRCA2 gene and how its failure might lead to breast
cancer.

But for the majority of my two-month stay, we attempted to
make antibodies that would latch onto the protein specified by
the BRCA2 gene. These antibodies will act as markers to help
researchers track how that protein functions, and how a
mutation in it might lead to breast cancer.

Great Expectations

Having no experience in a professional research laboratory
before last summer, I came into the program unsure of what to
expect. The only labs I had ever seen were either on television
or in the movies. But Hollywood's depiction of the lab is
undoubtedly based more on entertainment value than on fact. We
are all familiar with the endearing story of the

absent-minded professor and his wacky, accidental invention,
flubber. And who could forget the stereotypical mad scientist
scurrying around an eerie cellar filled with flasks of bubbling
liquids and peculiar machines that crackle with
electricity?

Yet, even though I had discounted those stories as fiction,
to some degree I still hoped to find an eccentric gray-headed
scientist busily mixing chemicals in his cluttered lab, tucked
away in a corner of the beautiful neo-gothic Medical Sciences
Research Building. I expected an eight- week experience filled
with exciting discoveries and non-stop action.

Almost every day, headlines in the nation's newspapers
detailed the latest break-through in cancer research. Maybe
there would be a headline that summer that would read: "Duke
University Lab Pinpoints Root of Inherited Breast Cancer."
Feeling the prestige of a veteran researcher with my white lab
coat and official Duke ID, I knew an amazing discovery was just
waiting for me around the corner.

Then again, maybe not. As I set foot into the modern,
immaculate lab, I found a calm methodical atmosphere. An
M.D.-Ph.D. student stood at her bench carefully pipetting
samples into tiny tubes for a PCR reaction, as a laboratory
assistant leaned quietly over a deep sink on the opposite wall
washing equipment. In the next room a small figure peered into
a microscope reviewing the immunohistochemical slides she had
made, while in the tissue culture room next door the lab
technician worked with Petri dishes containing a couple of cell
lines.

I must have taken a wrong turn. This was not the lab I had
ordered in my dreams. Where were the exclamations of Eureka!?
Where was the energy-charged bustle of activity? Here was only
an atmosphere of routine procedures in rooms filled with quiet
thought.

Over the two months' period, however, I became intimately
familiar with routine procedures and pensive quiet. I cannot
begin to count the number of DNA samples I must have prepared
for PCR or gels for a protein separation process known as
electrophoresis. And certainly the exposure to basic lab
techniques will give me an advantage in future biology course
labs. But, it was the incidental lessons in immunology and
pathology and the opportunity to use analytical skills that
made the program a meaningful educational experience.

My labs in high school had always taught concepts through
prescribed "cookbook" procedures, but our research required us
to draw on concepts I had previously learned to choose
procedures that would deliver answers. So, even though the
research lab may not present the surface picture of non-stop
action we often envision, I found it to be much more exciting
than it may appear to the untrained eye--the eye through which
I skeptically gazed that first day in the lab.

Lords of Discipline

Precision. That's the other difference between student and
professional labs--the scientific community demands it. And
that means devoting the utmost attention to avoiding
contamination in preparing procedures and maintaining cell
lines. For those unfamiliar with sterility practices, that may
seem like a simple task. However, as I quickly learned, sterile
technique requires an often underestimated degree of skill.

So, while gripping a container with the bottom three
fingers, I had to twist off the lid with my index finger and
thumb so that my other hand could carefully insert a pipette,
fitted with a small disposable tip, into the solution, without
touching the sides of the container or immersing the pipette
too far into the solution.

Then I had to pull the pipette out with just as much care,
locate the target tube, uncover its lid and deposit the liquid
as quickly as possible so as not to risk contamination by
air.

I can only guess at the outrageous number of pipette tips I
must have used to guarantee purity in the hundreds of such
transfers I had to do. As I quickly learned, PCR is undoubtedly
one of the most tip-consuming procedures to prepare for.
However, as I should have quickly learned, PCR also requires an
even higher level of sterility, accomplished in part by tips
stuffed with sterile cotton. But I didn't. Once, as I was
pipetting the last drops of PCR enzyme into the final tube, my
research partner pointed out the unstuffed tip on the end of my
pipette. In silent frustration, I turned my eyes to the box of
stuffed tips in front of me and resolved to do the painstaking
process once more–this time with more care.

The Time Machine

Repetition, especially of procedures, is not the only thing
a novice like me notices in the lab, however. Though time may
be "of the essence" in the race to find cancer cures, lab
procedures certainly take a lot of it, and not in a regularly
paced way. I experienced blocks of hurried preparation, often
for two procedures at a time, separated by seemingly endless
stretches of waiting. In many cases, hours pass before the next
step in a procedure.

And just what does one do with all of that time? On the
surface, it may seem to be a prodigious waste of a day. But it
was during those pauses that Duke's role as a teaching
institution became unmistakable. Dr. Hale often took those
times not only to evaluate our progress but also to explain the
cellular mechanisms at issue in our research and to share
slides from her most recent presentation.

One day, as I passed by another lab hosting a student for
the summer, that researcher even invited me to listen to an
impromptu lecture about her work with bone marrow.

At other times, I had the opportunity to visit Surgical
Pathology where tissue specimens are processed into slides for
microscopic evaluation. And on several occasions, I attended
the Autopsy Conference in Duke South Hospital to view organs
recovered post-mortem from cancer patients.

But it was my visits to patients in the breast cancer clinic
that brought my experiences of the previous eight weeks into
dramatic perspective. Like my mother, these patients are
fighting against a treasonous cell that has made them hostages
in their own bodies. Their only hope, it seems, lies in the
answers produced in research laboratories distant from their
hospital rooms.

At first, I had seen my laboratory work as just an abstract
business of experiments and results, but that clinic visit
helped me realize that research has many human faces--not only
the faces of the patients in the waiting areas, but the doctors
in the examining rooms, the researchers in the lab, and even
the students like me spending a summer observing it all.

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