When I fantasized about places that I would spend my summer before my senior year in college, I always envisioned myself on the coasts of Italy or relaxing on a Spanish beach. However, when my faculty sponsor, Dr. Steve Stratton, suggested that I consider enrolling in the BRAVO! Program I could not resist the opportunity. I carefully considered my options. I was not completely sure that I could give up the scorching sun, the monsoons, or the third-degree burns received when entering my car for an entire three months! I ultimately decided to join the laboratory of Professor Dr. Frederick Charles Campbell and Dr. Gavin Collett in the Department of Surgery at The Queen’s University Belfast in Northern Ireland. By the end of the three months I realized I had made the right choice. Not only because the warmest the temperature reached in July was 74 degrees F, but because the experiences I gained in the laboratory were invaluable.

For three months, I immersed myself in scientific research investigating the mechanism of action of the chemopreventive and natural product, curcumin. Previous research from Dr. Gavin Collett has demonstrated that curcumin-mediated apoptosis is induced by c-jun N-terminal kinase (JNK). However, the key genes that are activated by JNK which result in an induction of apoptosis have not been identified in human colon cancer cells. My project was designed to identify if two of the possible genes are key in this particular pathway. Those two proteins were bim and FasL. This study was necessary and important in establishing the roles of these proteins in connection to the mechanism of action of curcumin. An increased understanding of how curcumin induces apoptosis may enable its use in therapeutic strategies as well as in chemoprevention.

Normal levels of protein expression of fas ligand and bim in both untreated and curcumin treated HCT116 cells were analyzed by western blot at various timepoints. The technique of western blotting was not new to me; however, the most difficult aspect of my research was to draw conclusive results from the western blots repeatedly. Having to optimize everything from the amount of sample loaded, to the percentage of gel ran, to the dilution of the primary and secondary antibodies, was extremely demanding. Realizing that this meticulous way of attaining information is the essence of scientific research, I now look at my frustration with western blotting techniques as an extremely valuable lesson in persistency and patience. In the end, I managed to attain several conclusive data, although there are still some that will have to be repeated in Dr. Campbell’s laboratories.

At the same time the levels of mRNA expression of fas ligand and bim in untreated and curcumin treated HCT116 cells using RT-PCR methods also were conducted. Also, using a pharmacological inhibitor of JNK and conducting an RT-PCR on those RNA extracts to find any changes of expression when compared to curcumin treated cells alone will determine the dependence of fas ligand and bim on JNK. The evaluation of mRNA was much easier to complete than the protein evaluation of the proteins. Although there were other fallbacks encountered, like the machine breaking and being out of commission for two weeks, I was able to complete the data needed to evaluate the proteins on an mRNA level. While three months in any other field may be more than sufficient time to complete a project, it is barely enough time in scientific research to set things up for most projects.

I was very glad that I also had the opportunity to immerse myself in the unique culture of Northern Ireland. Belfast is unlike any other city in Northern Ireland because it has attitude, a certain edge that cannot be ignored. Although the religious tensions are not as palpable as they were a few years ago, small groups and areas can be found that cannot move past that point of their history. I became accustomed to the North Irish manner of speech, fast and harsh as it is, the infamous pub life as well as tea times. I visited the greenest land I have ever seen in my life while I was in Northern Ireland. The North Coast and loughs found within the country are incredible views in themselves. I also made a visit to the Republic of Ireland and saw the major differences between the North and South that I would never have knew existed if not for this opportunity. In the South, the accent is softer and life is much more easy-going. I even made it to the continent a few times to visit Spain and France.

Although, the time I spent working in the lab was invaluable to me, I will cherish these memories forever, partly because it enabled me to grow as a person as well as a future scientist. The educational experience was immense and satisfying in itself. However, the cultural, personal and social lengths that I have grown into allowed me to be independent in a foreign country, as well as made me adaptable and self-reliant -- qualities that are key to my life as well as to my educational experience. I implore all who are considering becoming a part of the BRAVO! program to do so because it truly is a once-in-a-lifetime experience that is immensely fulfilling. I would like to thank my faculty sponsor, Dr. Steve Stratton, my BRAVO! faculty sponsors Dr. Gavin Collett and Professor Dr. Frederick Charles Campbell as well as BRAVO! Director, Carol Bender for making this unbelievable experience possible.

Aluvia Escalante, UBRPer in Dr. Steve Stratton’s lab, Department of Medicine, Hematology/Oncology