Yavuz Dodurga¹, Cigir Biray Avci², Sunde Yilmaz², Zeynep Özlem Dogan Sıgva², Nezih Oktar³, Cumhur Gunduz²
¹Department of Medical Biology, Medical School, Pamukkale University, Denizli, Turkey
²Department of Medical Biology, Medical School, Ege University, İzmir, Turkey
³Department of Neurosurgery, Medical School, Ege University, İzmir, Turkey
Cyclosporin A (CsA) is a powerful immunosuppressive agent. MicroRNAs (miRNAs) are a class of recently discovered non-coding RNA genes that post-transcriptionally regulate gene expression. It is aimed to investigate the effects of CsA on the potential 88 miRNA expression changes in glioma cells-U87-MG.
U87-MG cell line grown in BIOAMF-1 medium supplemented with 10 000 U/ml penicillin, 10 mg/ml streptomycin, 2 mM L-glutamine was maintained at a density of 5 X 105 cells/ml. CsA was used in treatments of 10, 30 and 60 μM. Cytotoxic assays and determination of IC50 dose of CsA in U87-MG cells were performed. The cell group that was not treated by any agent was approved of the control group. Relative quantitation of 88 miRNAs was measured by Light Cycler 480 Real Time PCR. SNORD48, SNORD47, SNORD44, U6 were used as human endogenous controls. The fold changes of miRNAs determined and alterations in the miRNA expressions were compared with CsA treated and CsA- free U87-MG glioma cells.
In U87-MG cells treated with CsA (10 μM), 3 of 88 human miRNAs were up-regulated and 40 were down-regulated as detected with the miRNA array compared with control group. It is found that expression levels of several miRNAs, in particular, miRNA-195, was significantly decreased in CsA treated U87MG cells.
The study can provide important roles of miR-195 in GBM pathogenesis and in the molecular etiology of GBM. Rather than knockdown of miR-195 for moderate cell killing effect, treatment with CsA could be more effective especially on temozolomide resistant cells.
Keywords: Cyclosporin A, U87-MG, miR-195