Turkish Neurosurgery 2020 , Vol 30 , Num 2
Inhibition of the Invasion of Human Glioblastoma U87 Cell Line by Ruxolitinib: A Molecular Player of miR-17 and miR-20a Regulating JAK/STAT Pathway
Emre DELEN1,Oguzhan DOGANLAR2,Zeynep Banu DOGANLAR2,Ozlem DELEN3
1Trakya University School of Medicine, Department of Neurosurgery, Edirne, Turkey
2Trakya University School of Medicine, Department of Medical Biology, Edirne, Turkey
3Trakya University School of Medicine, Department of Histology and Embryology, Edirne, Turkey
DOI : 10.5137/1019-5149.JTN.26122-19.1 AIM: To determine the interaction between ruxolitinib, JAK/STAT signalling, and two angio-microRNAs (miRs) to expose potential target molecules in the inhibition of glioblastoma invasion.

MATERIAL and METHODS: The invasion properties of glioblastoma were analyzed using a cancer cell spheroid invasion assay. Following treatment of 195 nM ruxolitinib, the relative expression levels of miR-17 and miR-20a and genes of IL-6/JAK/STAT3 receptor signaling belonging to the JAK/STAT pathway were measured by qRT-PCR in treated and untreated three-dimensional tumor spheres of U87 cells.

RESULTS: Our results indicated that a therapeutic dose of ruxolitinib (195 nM) significantly increased miR-17 and miR-20a expression. Ruxolitinib treatment resulted in the production of IL-6 and active formation of IL-6 receptor complex for the subsequent activation of the IL-6R/JAK2/STAT3 axis. However, ruxolitinib treatment significantly decreased the expression of JAK2 and PI3K. Pearson correlation analyses revealed a strong negative correlation of miR-17 with JAK2, STAT3, and PI3K expressions, and also miR-20a has a negative correlation with expression levels of JAK2 and PI3K. The only positive correlation was found to be between miR-20a and IL-6, gp130 expressions.

CONCLUSION: The specific JAK2 inhibitor ruxolitinib plays an important role in glioblastoma angiogenesis biology via inhibiting IL-6 receptor-dependent JAK/STAT signaling. Additionally, both miR-17a-3p and miR-20a overexpression induced by ruxolitinib treatment may be playing a major role in downregulated JAK2, STAT3, and PI3K proteins. Our results suggest that miR-17-3p and miR-20a-5p may serve as new therapeutic targets for the treatment of glioblastoma. Keywords : Angiogenesis inhibitors, Drug effects, Glioblastoma, Invasion, Ruxolitinib, JAK/STAT signaling pathway, microRNAs, Molecular targeted therapy

Corresponding author : Emre DELEN, emredelen1979@yahoo.com