Effect of signal transducer and activators of transcription 3 on migration of human glioblastoma cells

Abstract

Objective To investigate the role of signal transducer and activators of transcription 3 (STAT3) signaling pathway in glioblastoma cell migration.Methods Janus kinase 2 selective inhibitor AG490 was used to inhibit the activation of STAT3 signaling pathway.The expression of STAT3 and p-STAT3 proteins in glioblastoma LN229 cells was detected by Western blotting.The optimal concentration of AG490 which can inhibit the activation of STAT3 in LN229 cell was chosen by methyl thiazol tetrazolium (MTT) assay.The migration ability of LN229 cells was evaluated by using wound-healing assay and chemotaxis assay.The cellular actin polymerization in LN229 cells was detected by the F-actin polymerization assay.Results When the LN229 cells were pretreated with AG490 at 50 μmol/L,there was no cytotoxic against LN229 cells ( t =1.862,P ＞ 0.05 ),however,the activation of p-STAT3 was suppressed ( t =4.821,P ＜0.01 ).The relative intensity of p-STAT3 was 0.291 ±0.036 in the 50 μmol/L AG490-pretreated cells and 0.530 ± 0.040 in the control cells.At 24th h after wounding,the migrating distance of the 50 μmol/L AG490-pretreated cells was (0.150 ± 0.054) ram and that of the control cells was (0.290 ±0.049) mm (P ＜ 0.05 ).After administration of 50 μmol/L AG490,the motility ability of LN229 cells was decreased in contrast to the control group in the chemotaxis assay ( P ＜ 0.05 ).When the 10 μg/L EGF was added in the low well,the trans-membrane cell number in the 50 μmol/L AG490-pretreated group was 24.52 ± 3.04 and that in the control group was 93.92 ± 9.52 respectively.Meanwhile,F-actin polymerization of LN229 cells was also significantly impaired with AG490 treatment ( P ＜ 0.05 ).Conclusion STAT3 signaling pathway plays a critical role in migration of glioblastoma cells.Inhibition of STAT3 signaling by AG490 treatment suppresses the migration ability of glioblastoma LN229 cells. Key words: Signal transducer and activators of transcription 3 ; Glioblastoma; Cell migration