My Internship
Overview
My internship is at the IU School of Medicine in the Pulmonology Department where I work under Dr. Sears in her lab. The main goal of my internship is to research the development and treatment of lung cancer in regards to a certain gene expression. This is important because nearly 34.1 million adults in the United States smoke. The research we do in this lab directly has an impact on the citizens around us!
Summary of Internship
Smoking induced lung cancer affects 90% of men and 70 – 80% of women, making it the leading cause of all cancer related deaths. Our focus is squamous cell carcinoma and how altered DNA repair contributes to development of early cancerous changes in epithelial cells. XPC is essential to genomic nucleotide excision repair (NER) for removal of bulky carcinogen – DNA lesions. Low expression of XPC has been observed in human lung cancers and mice with lung adenocarcinoma. Our laboratory found that mice deficient in XPC were prone to develop lung squamous cell carcinomas following exposure to the carcinogen NTCU. We hypothesized that XPC deficient mice treated with NTCU would develop pre-malignant changes in the pulmonary epithelial cells earlier than NTCU-treated XPC proficient mice. Using immunofluorescent staining for CK 5/6, marker of malignant epithelial cell transformation, airway epithelial cells of mice 8-, 10-, 12-, and 16-weeks after NTCU treatment were analyzed by airway size for percent CK5/6 positive. We observed increased CK 5/6 staining in lung squamous cell carcinomas, and in large compared to small airways at all time points, with CK 5/6 staining increasing over time. Ongoing research involves calculating whether XPC expression is associated with differences in CK 5/6 expression at different time points.
XPC at Work
Visual Aid
Here to the left is a picture of a labeled diagram of the structures inside a lung. As early, as 8 weeks we can see the staining of the CK 5/6 within the trachea. During 12 weeks there is some staining within the large airways at the segmental bronchus. At around 16 weeks there is significant staining in the segmental bronchus area and even near the smaller airways around the terminal bronchus. Our hypothesis is that pre-malignant squamous cells originate in the more central airways.
Each slide is cut at different angles which can include a combination of large, medium, and small airways.
A Look at My Current Work
Our site specifically looks at how the XPC gene is involved in DNA repair which affects the development of lung cancer within smoking patients. This is relevant research because some people may be more susceptible to early cancer than others depending on who exhibits this gene. In order to test this, we used mice who have shown similar correlations between the XPC gene and premalignant tumors. Over the course of my own research project, I have exposed mice deficient in XPC to NTCU and mice proficient in XPC to NTCU. I have then taken pictures of microscope slides of mouse lungs at 8, 10, 12, and 16 weeks after treatment to see if XPC deficient mice are more susceptible to develop premalignant changes. All the cells were stained with CK 5/6 which is a marker that identifies malignant epithelial cells. This stain will reveal if there is an increase in lung squamous cell carcinomas in XPC deficient mice. We have hypothesized that the trachea starts to show signs of cancerous cells as early as 8 weeks whereas the lungs show cancerous cells around 12 weeks. There are a few pictures below that are the first step in analyzing the data.
This is a picture of a mouse's trachea. It is important to remember to count only the epithelial cells within the picture. This specific one has many of them! All of the things that look like yellow specs are actually numbers. This is technique used to count how many cells are within one single picture. This is important information to have so that we can analyze the ratio of CK 5/6 stained cells to not stained.
To the left is a picture of a mouse's trachea at 10 weeks after treatment. This is achieved through a staining process called DAPI. DAPI staining is a process that stains all the nuclei within a slide. This allows us to count the total number of cells since all lung squamous cells have a nuclei.
What even is staining?
It is a tool used to detect molecules of interest within a certain specimen.
Final Project!!
Final Poster Revised.pdfA Journal of Learning and Skills
At the start of this internship in September I have a few personal skills that I want to gain from this internship. I want to become familiar and proficient in using the high technology microscopes to help in my future. I also want to better my interpersonal skills and become more confident when talking to my peers and supervisor. I made a plan of action where I know the best way to improve in these skills is to have more practice. I try to meet with my principal investigator (PI), Dr. Sears, at least once every two weeks to establish my confidence.
It is now October and I have many exciting things to look forward to in my internship! Mostly, I am excited to learn about the lab environment and how everything comes together to add to the lab’s bigger picture. In terms of new skills, I would like to learn how a comet assay is completed, since it is something that I had never heard of until starting my internship. I know it is one of the more advanced lab skills with regard to carrying it out properly from start to finish. I would also like to continue developing skills to speak with my superiors. A goal is to reach the point where I know what they are discussing and offering my own input. Since I am still relatively new to the lab and have less biology knowledge at hand it is hard to understand everything the first time. I think this is something that will become easier as time goes on.
I am confident in my ability to be independent, prioritize, plan, and achieve the goals I have for the day. I have been able to complete a few assignments on my own which has strengthened my ability to be independent. I have been working with the lab technician the past couple of weeks which has contributed to my interpersonal skills. I am able to have conversation with her about what we are doing and how I can achieve it by myself.
Something that I would like to improve on is using my resources to confront challenges and take risks. I am still a little timid in lab when working with new procedures and processes. I would like to take those risks and just try to figure things out while knowing that it might not work out. I think I could start by being more hands on when someone is explaining a method to me. I could try it out for myself but still have someone nearby to make me feel more comfortable.
It is now November and I have been taking BIO-K103 this semester and have been learning about the different body systems. Learning about the integumentary system, the different types of skin tissue, and cells has been helpful for this internship. I now know what it means to be looking at simple squamous cells. This has allowed me to deeply understand the differences between all the cell shapes. I have been analyzing and taking pictures of trachea and lung cells. Being able to tell, understand, and evaluate the differences has been very helpful because I can now differentiate what I see when I analyze the pictures. As soon as we learn about the respiratory system, I am sure that it will positively impact my understanding of my internship. This internship has allowed me to gain a deeper understanding of my classwork because I have been able to put what I have learned into a real-world setting. I think the point of learning classroom material is to understand it and apply it outside the classroom. This internship has given me the perfect opportunity to do that.
I am also currently taking cell biology which has been enormously helpful in my understanding of my work. Going deeply into the functions of DNA, RNA, and repair systems has been critical to understanding my work. One of the main focuses of my internship is the XPC gene and how it functions in the DNA repair system. Having a strong background and understanding of DNA has allowed me to understand harder mechanisms in the body like the XPC gene. I am sure once I reach even more upper-level biology courses it will only make things easier.
It is now December which means the first semester is wrapping up and I think I have developed the ability to pick up a technique and figure out how to replicate it on my own. This is one of the things I was nervous about when I first started working. I have gained more confidence in my skills and how I can handle new situations. I have had great experiences throughout this semester. One of my favorites has been leaning to use the fluorescent microscope and watching the dissection of a mouse. Mastering the use of the microscope has helped so much with all the slides that we make and receive. The dissection was an interesting experience because I was able to see the physical impact on a mouse lung based on how it was treated. Seeing and identifying the tumors on the lung while also learning the anatomy of mice lungs has been very satisfying.
This work was important because lung cancer affects many individuals across the globe. To see how a certain gene can impact the development of lung cancer in mice and humans. It serves the purpose of adding to research about lung cancer and how we can try to combat it.
One of the most inspiring changes I saw in myself was how I was able to become a better problem-solver and have more perseverance when things got difficult. I would like to continue to grow in how comfortable I am to independently create a plan based on some information. Since I have been working in the lab for only 7 months, I have started to finally understand the big picture of things and how my research fits into it. I think as long as I keep developing that understanding it will be easier for me to create my own plans without as much guidance from my supervisor.
Working with Professionals
Everyone that I have had the opportunity to work with in the lab is always finding new things to accomplish. They actively seek out new projects to start and share what they have been doing with each other. I think that is something that I want to do as a professional. I want to be someone who is always finding new things to work on and to best use my time.
I have also observed how comfortable some of the other graduate, doctoral, etc. researchers are talking to our supervisors about the work that they are doing. I would like to also become more comfortable and confident doing the same thing. When I first started working I could find it overwhelming to find the right words and questions to ask. After getting to know everyone better and working in the lab for a few months I have become more confident in my interpersonal skills.
Expectations vs Reality
Expectations Going In: I expected it to be work all day and always being busy with the lab work. I also expected to be expected to know how to do the different lab procedures. I soon found out that it really is not like that at all.
Reality: What I realized is that most of the time it is trying to figure out the meaning of the results that you get. I think that is the most difficult and time consuming part. Having an understanding of the work that I am doing is mostly what the internship is about. There is lab work most of the time, but that is fun and easier to replicate after learning it a few times. However, understanding the results is something that can be different each time.
Successes and Challenges
Success: I have had a lot of success in immunofluorescence staining and making sure my slides are properly prepared. I have had success in learning about new scientific studies related to our own field. I have had success in using the microscope and identifying the airways.
Challenge: The most challenging part of my experience was to figure out how to do things alone. After learning how to autoclave my dilutions once I had to do it myself and that was challenging. I got over this challenge by following the directions slowly and asking for help if I needed it. I plan on working on this in the spring by just doing it by myself more often and getting better at it.
Growth as a Professional
Over the past seven months, I have had a lot of exposure to professionals within the workplace. This has helped me develop my own idea of professionalism and the values within a workplace. It has influenced how I will continue to develop myself. For example, all the staff I have been with are very professional and willing to help each other achieve the common goal. Everyone is also so eager to learn what you are working on; it is evident that their professionalism allows the work environment to be successful. At first, I felt intimidated to talk to professionals who had much more experience than I had, but I have experienced something different. Everyone is willing to take the time to give me advice and explain whenever I have questions. They are excited to have questions and spend time explaining something which is not what I expected. As someone observing this professionalism, I have seen that being a professional is about being as knowledgeable as welcoming to others.