Analyzing the DNA Stability of U87 Glioblastoma Cells After Co-Culturing With HL-60 Neutrophils and Jurkat T-Cell Conditioned Media

Abstract

Glioblastoma (GBM), a Grade IV brain cancer, is aggressive, yet unfortunately common. As conventional treatments open fail to eradicate GBM and prevent recurrence, more-advanced immunotherapy, which has the natural advantages of immune cells against threats, has been considered as a prospective treatment option. However, in order to develop effective immunotherapeutic strategies against GBM, it is imperative to analyze the individual effects of the many immune cells first, such as the effects of T-cell secretomes and neutrophils. Using co-culture methods, U87 GBM cells were grown with the two immune treatments and harvested in 24 hour intervals three times. Their protein contents were extracted, quantified with bicinchoninic acid (BCA) assay, and the deoxyribonucleic acid (DNA) damage marker γH2AX was relatively quantified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western bloting. As DNA damage was thought to be an indicator of cancer cell death, the typically-an -cancer T-cell conditioned media (TCM) was hypothesized to promote DNA damage and the typically-pro-tumor neutrophils were hypothesized to suppress DNA damage. However, the acquired western bloting results seem to suggest otherwise, with the former having no significant effect at all and the latier significantly promoting DNA damage a tier 72 hours of co-culturing. Mistakes such as poor western blot quality or the misinterpretation of the meaning of DNA damage in immunotherapy could have affected the implications of the study, hence more research is required to form a solid conclusion to the effects of TCM media and neutrophils on GBM cells.