Cell and Molecular Methods

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  Lab Rats

  Samantha Aragon, Cristina Herrera, Felicity Rodriguez, Riley Smith, and Terry Shackleford

  Ewing sarcoma is an aggressive cancer that affects children & adolescents. Common treatments include surgery, radiation, and chemotherapy, yet individuals have a poor survival rate and often develop side effects from treatment, including fertility issues, secondary cancer, heart problems, and growth abnormalities.1SOX18 is a gene identified to aid in cell proliferation and cell differentiation, leading to the development of several cancers, such as gastric, bladder, and lung cancer.2,3 SOX18 resides in the JAK2/STAT3 pathway as a regulator, along with another gene named KIT that instigates the formation of the cell signaling pathway.4KIT is a transmembrane tyrosine kinase, which instigates cell growth, differentiation, and survival.5,6Thus far, no relationship has been stated between SOX18 and KIT. Due to shared presence in the same pathway, this project proposes that when KIT is knocked out, then EWS cells will have an increased expression of SOX18, leading to increased cell growth and proliferation.

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  Test Tubes

  Victoria Castillo, Melany Cervantes, Nicole Robles, and Terry Shackleford

  SOX18, a transcription factor with a DNA-binding HMG domain, plays a critical role in regulatory processes linked to cancer progression. Exhibiting oncogenic properties, SOX18 has been linked to various cancers, where it promotes tumor growth by enhancing cell invasion, uncontrolled proliferation, and resistance to apoptosis through dysregulated signaling pathways. STAT1, a protein crucial for immune system regulation, is prominently expressed in immune-associated tissues like the lymph nodes, and bone marrow, suggesting it could play a role in tumor growth. Ewing Sarcoma (EWS), an aggressive cancer that targets bone and soft tissue, carries a high mortality rate of 90% without treatment and 75-85% even with standard therapies, with a substantial risk of relapse. Given the involvement of SOX18 in oncogenesis and the immune-regulatory role of STAT1, we hypothesized that SOX18 expression would result in increased STAT1 and that knockdown of STAT1 would reduce cell viability and increase apoptosis. This would suggest that SOX18 contributes to the survival and proliferation of EWS cells. Furthermore, if SOX18 is overexpressed into the cell line of STAT1, which is associated with necroptosis, it may decrease cell viability and produce more cancerous cells.

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  Fig Neutrons

  Allie Woods, Samuel De La Cruz, Catherine Garcia, and Terry Shackleford

  Multiple experiments were conducted to measure the effectiveness of certain genes on cell growth. In the exploration of this process, RAC3 acted as the target gene and was applied to both SOX18 overexpressing and normal expressing cells (control). RAC3 is a gene that typically controls the intercellular signaling pathway of a cell. This means it helps with the movement of cells and allows for migration.The purpose of these experiments were to determine how the gene RAC3 would work when introduced to carcinogenic cells, specifically Ewing Sarcoma. Typically, RAC3 acts as a messenger for the cell, but when in conjunction with cancer cells, it acts as pathway that controls cell growth in response to disease. By Targeting RAC3 inside of the ES cells that are overexpressed with SOX18, there should be an efficacy in the knockdown of the overexpressing genes. Thus, Ewing Sarcoma will no longer be able to produce cancer causing cells.