Hodgkin’s lymphoma is a certain lymphatic disease caused by cancerous lymphocytes, which has lost its immuno activity through mutations. The cancer cells can uncontrollably proliferate towards such a high concentration that the healthy immune cells drown out, and the patient becomes at risk of infections. Though Hodgkin's lymphoma is treatable with cytostatics and/or radiation therapy, some patients suffering at an advanced stage of the disease do not recover from the treatment. This brings out the importance of pursuing a better understanding of both the disease and the treatment methods.

The purpose of this project was to study how the RNA expression in cancer cells of the type Hodgkin’s lymphoma (L-428 cells) are affected by the cytostatics vinblastine and doxorubicin. The goal was to be able to answer the questions at issue - “Which genes are up or down regulated in the surviving cells?” and “How are they affected and responding to the cytotoxic drugs?”.

A cell culture of human L-428 Hodgkin’s lymphoma cells was obtained and treated with the cytostatic drugs doxorubicin and vinblastine. Samples were taken from the culture 6 and 24 hours together with 1 week after the treatment. From the samples, RNA was extracted. A library preparation was performed and the samples were then sequenced. The sequence data was quality controlled, trimmed and mapped to identify the up-and down regulated genes of the treated cells in reference to the non-treated cells. Correlation- and DE plots were made, and the gene regulation data was applied to the website geneontology.org to analyze which biological pathways were most likely affected by the cytostatic treatment.

Gene ontology for the 6 hour doxorubicin treated cells mainly showed results regarding mitosis and cell division pathways, while the corresponding gene ontology analysis showed no results for the vinblastine sample.

Gene ontology for the 24 hours doxorubicin and vinblastine treated cells showed higher endoplasmic reticulum stress, activation of apoptosis pathways and up regulation of cholesterol biosynthesis. The cells treated with doxorubicin showed more results focusing on energy production and DNA repair, whilst vinblastine treated cells showed a higher focus on stress, apoptosis and MAPK impact.

Gene ontology for the recovery samples of doxorubicin showed 4 pathways that were upregulated. Lymphocyte migration, lymphocyte proliferation, cadherin binding and tumor necrosis factor (TNF). The first 3 genes result in an increased risk of metastasis while TNF increases the survivability of the cell by upregulating cell growth. This makes these four genes a topic of interest.

Gene ontology for the recovery samples of vinblastine showed T cell apoptosis genes were upregulated, including the gene for CD274, which inhibits T cell activation and cytokine production. A result regarding cytokine response was also obtained, which correlates well with the CD274 result. Also, genes involved in T cell migration, differentiation and lineage were upregulated, which can indicate a positive response to the treatment. Results regarding peroxidase activity and haptoglobin-hemoglobin complex was also obtained.