Student Research at Duke

Looking back, I couldn’t be happier that I decided to dedicate my summer to exploring a very probable future career.  In no other way could I have seen the inner workings of science without the glamor of TV or the insufficiencies of personal description. 

From having a day off to slaving in the lab until 11 at night, from idly browsing through relevant articles to rushing to and fro between 3 experiments, I experienced the total lab package.  I formed a bond with my labmates and a playful sense of competition with the neighboring lab.  Most importantly, I gained an understanding of research as it exists today: western blotting, immunofluorescence under the microscope, cell cultures, ligations, you name it.

If I had a second chance to decide my plans for Summer 2007, I wouldn’t hesitate to choose Howard Hughes Research Fellows all over again.

Having heard from a variety of students, either attending grad school or an MD/PhD program, I am quite sure that grad school is in my future.  I can certainly see myself working in a lab for some period of time, though it seems I may tire of it in several years’ time.  However, with a more comprehensive understanding of cell biology, biochemistry, or whatever I choose to go into should come a heightened sense of enthusiasm that could be the driving force behind a lifetime of research.  Just in case I do tire of such a career, the input of the career panel suggested that a broad range of jobs can be pursued with a PhD in science, further assuring my decision for grad school is a good one.  As for the schools to which I’ll apply and hope to attend, I have 3 years to think about that.

Having heard from a variety of students, either attending grad school or an MD/PhD program, I am quite sure that grad school is in my future.  I can certainly see myself working in a lab for some period of time, though it seems I may tire of it in several years’ time.  However, with a more comprehensive understanding of cell biology, biochemistry, or whatever I choose to go into should come a heightened sense of enthusiasm that could be the driving force behind a lifetime of research.  Just in case I do tire of such a career, the input of the career panel suggested that a broad range of jobs can be pursued with a PhD in science, further assuring my decision for grad school is a good one.  As for the schools to which I’ll apply and hope to attend, I have 3 years to think about that.

In order to discuss the status of my research, let me first explain the objectives of my research.                                                                                                                             
In Dr. Seewaldt’s lab, one of the major focuses centers around finding biomarkers for breast cancer, whether they might lead to preventative treatments or else a diagnosis.  A protein called PELP1 holds promise for providing us with such a biomarker.  PELP1 is a coregulator protein of estrogen receptor (ER) and usually performs its regulation tasks strictly in the nucleus.  However, it’s been recently discovered that certain cancer cell lines contain high amounts of cytoplasmic PELP1.  Here’s where our research comes into play.     
ER, which is typically found in very low levels in normal human mammary epithelial cells (HMECs), is frequently used to identify the pre-cancerous condition, atypia.  Women with atypia are sometimes given the option of Tamoxifen treatment in order to prevent the progression to cancer.  However, not all women respond to this drug, which has many unpleasant effects such as the increased risk of endometrial cancer.  It is suspected that cytoplasmic PELP1 may be present in those women who are resistant to Tamoxifen, activating certain pathways that lead to cell proliferation and hormonal independence.  Furthermore, the harder-to-detect ER-negative forms of atypia and breast cancer cannot be identified based on ER levels; the ER levels appear normal, yet the cells proceed to proliferate and become cancerous.  If we discover cytoplasmic PELP1 in such atypical cells, perhaps the protein can be used as a more efficient detection technique.                                
The first step of our research is to confirm that the mutant PELP1-CYTO plays a part in activating the cancer-promoting AKT and MAPK pathways in ER+ cell lines.  Similarly, we want to see if PELP1-CYTO can exist in and, if so, activates the same pathways.  Next we’ll test if the protein can be transfected into and have the same effect in normal HMECs.  This will tell us if such fluctuations in PELP1-CYTO are evident before cells actually become cancerous.  With later investigation, we’ll take actual RPFNA (an aspiration technique for sampling cells from high risk women) samples and look for evidence of PELP1-CYTO and whether or not it correlates to resistance to Tamoxifen.                           
Currently, we’ve run one western blot to confirm the activation of AKT and MAPK pathways.  However, results were not definitive.  We’ve also performed immunofluorescence with an ER+ breast cancer cell line to experiment with the conditions needed to detect PELP1 most efficiently.  All other efforts (digests, transfections, ligations, western blotting) have been aimed at preparing for future procedures.  By the end of this program, we hope to have determined the effects of PELP1-CYTO in ER+ and ER- breast cancer cell lines as well as in normal HMECs in in vitro models.

In response to the article, “An Unwelcome Discovery” that I read and the movie, “And the Band Played On” that we watched, I can only say that I am thoroughly disgusted by the lack of integrity displayed by these head researchers.  The spirit of research lies in the desire to investigate, predict, and deter conditions affecting the human race.  Without such a desire, one might as well turn to mowing lawns as a livelihood.

With this being said, it’s such a shame that investigators like Eric Poehlman and Robert Gallo damage society’s perception of science through plagiarism and fabrication.  It’s bad enough that many misconstrue investigative inquiry as immoral, or believe the intricacies of life incomprehensible to any being less than divine.  In a day that exaggerated media coverage can destroy the reputation of innocent individuals or institutions, the Poehlmans and the Gallos of the scientific community risk the credibility established by researchers over the centuries.                                                                                                                
In reference to Dr. Dave McClay’s lecture, I now comprehend how laziness and carelessness could play a distinguishable role in research quality.  Without the utmost attention to one’s experiment, the smallest of inconsistencies could skew results, and frequent repetitions of such practices could sabotage an entire project.  Although I take every message that has been thrown at us throughout this “ethics week” with all seriousness and deliberation, I will view the latter as the real take-home message.  I just don’t understand how a person could be tempted to invent or steal data with such dishonorable motives and foul implications that will follow otherwise.  Thus, I will focus on performing even the most minor of tasks in the lab with the utmost attention. Now if only that could negate my clumsiness….

Upon interviewing my postdoc mentor, I found that she had taken a straightforward approach to getting where she is.  College with undergrad research, grad school (at UNC, explaining the occasional derogatory Duke remark), and a couple of jobs as a postdoc.  What I was fascinated, and sort of relieved by, was Julie’s explanation that medical school never interested her in the least.                                                                                  Having a father who’s the CEO of my hometown’s small hospital, I hear every night about all of the feuding between doctors and patient complaints about doctors and doctors’ being on call every waking hour for a week.  I understand the huge rewards that can come with being a doctor, but I think laboratory research can have farther-reaching rewards given time and dedication.                                                                                                          Coming into Duke, I thought and hoped that I would find multitudes of scientists, students hoping to pursue the field for the sake of furthering current knowledge.  But instead, I found pre-meds.  I’m taking physics… because it’s a pre-med requirement to struggle through?  No, because it’s an interesting, useful course, believe it or not.  If I tell someone I’m a chemistry major, I’m automatically asked if I’m pre-med.  It seemed that scientist and pre-med were synonomous around here.  Making matters worse, I feel like the underachiever in this situation only because I’m opting not to go to med school.  I feel like I could get accepted somewhere and do moderately well in med school, but I simply don’t won’t to be a medical doctor.  I like school, and hope to continue taking classes as long as I can possibly get away with it, just not with the expectation to end up as an M.D.                                                                                                                      Anyway, hearing that my mentor was never tempted by society’s seemingly high profile career as an M.D. was welcome news to my ears.  I just needed to know that there are others out there who can dedicate their lives to science and feel entirely fulfilled with what they do.

Now that my mentor is back in town and recovering from her back injury, I’ve been a lot busier in the lab.  Since getting to work, I’ve seen a lot of and done a little Western blotting, where we run proteins via electrophoresis and transfer the protein bands to a PVDF membrane.  Once on the membrane, the proteins can be tagged with an antibody, which is in turn identified by a secondary antibody.  This secondary antibody is receptive to a solution called Super Signal that, upon mixing, luminesces and can be captured on an imaging device.  In this way, we can test for specific proteins, seeing if they exist equally in normal human mammory epithelial cells (HMECs) and in various cancerous cell lines.                                                                                                                          So far most of my time has been spent focusing on this particular technique.  First we started with smaller, pre-packaged gels, but now I’m learning how to pour and set my own larger gel.  I’ve also been learning about mixing and preparing all the solutions and samples needed for the procedure.  I’ve also worked on bits and pieces of other experiments that my mentor has underway.  As my research question becomes more well-defined, hopefully I’ll learn more about these various lab techniques.  As for now, I must return to my lab for another day of discovery.

 This is the lab where I’ll be doing most of my work.  And here you can see some of the gels that others in this lab are running:                                                                                                            Unfortunately Julie, my mentor, has hurt her back after returning from a trip, so I’ve only been reading some more articles and watching others at work.  Hopefully I’ll be able to get to work next week!