Discipline: Biological Sciences
Subcategory: Cancer Research
Jocelyn Ricard - University of Minnesota - Twin Cities
Co-Author(s): Logan Spector, PhD; University of Minnesota, Department of Pediatrics, Division of Epidemiology ; Erica Langer; University of Minnesota, Department of Pediatrics, Division of Epidemiology
Background and Aims: Mutations in the β-catenin gene have been detected to lead to the contribution of the development of hepatoblastoma tumors. The aim of the research is to investigate effects of the candidate gene, CTNNB1, on Hepatoblastoma cell behavior. We also compared the motility of HepG2 and HOS cell lines in the Transwell Migration Assay and Wound Healing Assay. CTNNB1 lead to a gain of function advantage in hepatic cancers and that gain of function matters because it influences known invasive and metastatic pathways that involve cellular migration, growth, etc; which is we hypothesize that there will be a larger number of cells in the assays after removal of the CTNBB1 gene. Methods: We used CRISPR to knock out CTNNB1 (targeting Exon 4 to match SNP site) in HepG2 cells. The normal HepG2 cells, the gene edited HepG2 cells, and the HOS cells were compared in the Transwell Migration Assay and the Wound Healing Assay to see if there was a difference and if so, which cells show a greater count in the membrane, which shows us the motility of these cells. Both assays demonstrate migratory properties of cells, however, the Wound Healing Assay is far more complex and representative of cellular movement. Results: The results are in progress, but we expect to see more cells in the membrane in the Transwell Migration Assay after completion and greater cell migratory movement in the Wound Healing Assay with the gene-edited HepG2 cell lines. Overall, it is expected to see the HOS cells migrate more quickly through the motility assays. Conclusion: The HOS cell line had greater motility in the Transwell Migration Assay and the HOS cell line had greater motility in the Wound Healing Assay. Based on the pending results of the gene-edited cell line, the greater the number of cells in the membrane which gives insight into cancer cell movement and would give an idea of transformation with that genotype. The mutation could be a gain of function mutation which could promote proliferation of cancerous cells. Keywords: hepatoblastoma; osteosarcoma; CTNNB1; β-catenin; transwell migration assay; wound healing assay.Not Submitted
Funder Acknowledgement(s): North Star STEM Alliance - Louis Stokes Alliance for Minority Participation National Science Foundation
Faculty Advisor: Logan Spector, PhD, email@example.com
Role: I cultured the cells, performed the migration and wound healing assays, stained the cells to prepare for the assays, research experimental methods to give us the best outcome.