Blockage of the ROCK pathway inhibitsthe invasion and colony formation ability of glioblastomaspheroids.

Background: Glioblastoma is the most common and highlyinvasive central nervous system tumor. The Rho/ROCK signalingpathway is an important signal transduction system thatis critically involved in cell growth, differentiation, migration,and development and plays a pivotal role in the invasion andmigration of glioblastoma cells. Here we south to determinethe role of ROCK Inhibitor (Y-۲۷۶۳۲) on migration and invasionof glioblastoma cell line (U۸۷-MG) in both monolayer andspheroid models. Materials and Methods: Cells were treatedwith different concentrations of Y-۲۷۶۳۲ (۳۰, ۶۰, and ۱۰۰ μM).ROCK۱ & ROCK۲ expression levels were assessed by qPCR.viability of cells was assessed by MTT in monolayer and thegrowth rate of spheroids was also analyzed. Cell migration wasanalyzed by using scratch assay and disaggregation assay inmonolayer and spheroids respectively. Moreover, the effect ofROCK inhibitor (Y-۲۷۶۳۲) on U۸۷ colony formation was assessedby colony assay for ۱۰ days. A spheroid invasion assaywas performed using Matrigel and collagen. Results: ROCK۲expression was decreased following treatment of cells withY-۲۷۶۳۲. The dose of ۱۰۰ μM ROCK inhibitor had no significanteffect on cell viability and spheroid growth rate but a lowdose of Y-۲۷۶۳۲ (۳۰ μM) increased cell viability and growthrate of cells in monolayer and spheroid models respectively.Migration of cells in monolayer was increased in the presenceof a low dose of Y-۲۷۶۳۲ (۳۰ μM) compared to the control.Interestingly disaggregation of spheroids was increased after۴۸h in the presence of Y-۲۷۶۳۲ (low and high doses) relativeto control. Colony formation ability and invasion of cells weredecreased following the inhibition of the ROCK molecule.Conclusion:Altogether our data suggest that the inhibitory effect ofY-۲۷۶۳۲ on cell migration varies depending on the dose and thepresence of the extracellular matrix. Our data also proposed thatthe ROCK pathway may play a promoting role in glioblastomacell invasion.

کلیدواژه ها:

Glioblastoma ، ROCK inhibitor ، migration ، invasion ، spheroid ، Rho/GTPase