U87_Radio_Resistance_Invasive_Noninvasive

ABSTRACT: Background: Glioblastoma multiforme (GBM) is the most common and malignant primary brain tumor in adults. Radiotherapy is together with surgery the mainstay treatment in management of patients with GBM, however, infiltrative growth pattern and radioresistance of glioma cells impedes therapeutic effects. Our study sought to investigate 1) whether GBM invasive cells render them resistant to radiotherapy and 2) what are the potential molecular mechanisms underlying the radioresistance via gene expression profiling.
Procedures: In-vitro selection of invasive cells using matrigel-coated transwell assay: U87 cells were starved 8h prior to seeding. 5x105 cells in 2 mL serum-free DMEM medium were plated in the upper transwell inserts of a 6-well Boydem chamber. 10% FCS was added in the medium of the lower chamber. After 20h incubation, non-invasive cells on the surface of the membrane were scratched and recovered. Invasive cells migrating through matrigel due to the gradient of attractant were recovered by trypsin/EDTA after the membrane was washed with PBS twice and swabbed by cotton tips. Evaluate the invasiveness of selected phenotype of U87 cells by time-lapse wound-healing assay: 5x104 cells in 100μL serum-free DMEM medium were seeded in a 96-well ImageLock plate with each well was pre-coated with matrigel. Wounds were scratched using a 96-pin WoundMaker and gently washed with 2xPBS. After cooling equilibration, the wounds were coated with a second layer of matrigel. The microplate was incubated at 37oC and 5% CO2 for 30min. Cell invasion was recorded by living-image microscopy over 48h. Microarray: after confirmation of enhanced invasive ability, invasive and non-invasive cell populations were used for RNA isolation, cDNA transcription, and chip hybridization.

ORGANISM(S): Homo sapiens

SUBMITTER: Zili Tang 

PROVIDER: E-MTAB-4608 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress