High resolution analysis of anti-migratory inhibitors in high grade glioma treatment

Abstract

IntroductionGlioblastoma Multiforme (GBM) is an extremely aggressive form of glioma tumour. The characteristically invasive properties of GBM contribute to the poor prognosis in suffers. This study focuses on the therapeutic potential of four anti-migratory drugs - 6-bromo- indirubin-3’oxime (BIO), CCG-1423, Latrunculin A (LAT A) and Lithium Chloride (LiCl), in vitro. The migration distances of migrating GBM cells and cell circularity for each treatment and an untreated control were measured and statistically analysed.MethodsGBM spheroids of the U251 cell line were treated with BIO, CCG-1423, LAT A or LiCl plus an untreated control in preparation for this study. These were sectioned using immunohistochemical techniques and scanned prior to the study using QuPath. Cell migration was measured from the perimeter of the tumour spheroid to the edge of the migrating cell. Cell circularity was obtained using the ‘cell circularity index’ function. The cell circularity of the migrating cells was obtained separately to the cell circularity of the cells within the spheroid. This repeated for each section for each spheroid for each study. The data obtained was analysed in SPSS.ResultsEach of the treatments showed significant statistical differences compared the control spheroid for the migration distance measurements. LAT A was shown to have the greatest effect in decreasing the migration distances. The analysis of the cell circularity indicated that the migrated cells were significantly more rounded than the tumour cells for each treatment. The ‘cell circularity index’ indicated the migrated cells were more rounded than those within the tumour spheroid across each of the treatments.ConclusionThe cell migration distance study suggests further study, to analyse the effects of these drugs in vivo, as they each demonstrated anti-migratory effects on the GBM cells. The ‘cell circularity index’ study should be amended to more accurately represent the changes in morphology during GBM cell migration.