The Study Of Anti-tumor Activity Of STAT3 Inhibitor HJC0152 In Glioblastoma

Objective:To investigate the anti-tumor effect of Signal Transducer and Activator of Transcription 3（STAT3）inhibitor HJC0152 in human glioblastoma and provide a theoretical basis for the exploration of the therapeutic approach of human glioblastoma and application of HJC0152 in cancer treatment.Method:In this study,three human glioblastoma cell lines U87,U251 and LN229 were selected and treated with DMSO and HJC0152 as control group and experimental group respectively.The antitumor activity of HJC0152 was evaluated by comparing the two groups of cells after drug treatment.Experimental research is divided into experiments in vitro and in vivo.In vitro:IC₅₀ of HJC0152 was determined by MTT assay and used as the stimulating concentration.Expression level of STAT3,p-STAT3（Tyr705）,P-STAT3（Ser727）,epithelial-mesenchymal transition（EMT）related proteins,proliferation and senescence-related proteins and apoptosis related proteins was determined by Western bolt.Transwell assay was used to detect the mobility of cells.Colony forming assay was used to test cell proliferating ability.SA-β-gal staining assay was used to detect the aging state of cells.Flow cytometry was applied to examine cycle phase distribution and apoptosis rate.Distribution of Bax and cytochrome C were detected by Western blot.JC-1 probe was applied to detect mitochondrial membrane potential.In vivo:U87 was used to establish animal model.Mice were intratumorally injected with DMSO or HJC0152 daily.Body weight and tumor size were recorded.Tumor size and weight were compared to evaluate the suppressive effect of HJC0152on tumor growth.Result:HJC0152 induced significant decrease of p-STAT3（Tyr705）but hardly affected STAT3 or p-STAT3（Ser727）.The expression level of N-Cadherin,Twist-1,Vimentin,MMP-2 and MMP-9 decreased and the expression of E-Cadherin increased after HJC0152 treatment.HJC0152 attenuated glioblastoma cell ability of mobility,and induced accumulation of G1 phase cells,raised expression of p21 and reduced that of cylinD1,thus suppressed proliferation of glioblastoma cells.HJC0152 induced cell senescence and increase of p16 expression.Mitochondria associated apoptosis was observed following HJC0152 treatment.Mitochondria membrane potential declined and cytochrome C was found to escape out from the mitochondria and Bax to translocate the other way.HJC0152 sensitized glioblastoma cells to cisplatin assessed by MTT and colony forming assay.The animal model indicated that HJC0152 was able to suppress growth of glioblastoma cells in vivo.Conclusion:As an inhibitor of STAT3,HJC0152 exerts a potent anti-tumor activity in vitro and in vivo.This experiment provides a preliminary experimental basis for HJC0152to be the STAT3 targeted drug for treating human glioblastoma in the future.