The Effect Of Hydroxysafflor Yellow A In Glioma Via P38MAPK Signal Transduction

Objective To observe effect of Hydroxysafflor yellow A(HSYA)on proliferation,migration and invasion of human glioma U251 cell line.Detect expression level of protein related with P38 Mitogen-activated protein kinases(MAPK)signaling pathway and investigate its possible mechanism.Methods Glioma cells U251 were cultured in vitro and randomly divided into seven groups as follows:blank control group(without any drug),low,middle,high concentration positive control groups[100,300,900 μmol·L-1 temozolomide(TMZ)],low,middle,high concentration test groups(25,100,400 μmol·L-1 HSYA),combination groups using each concentration of TMZ combined with different concentration of HSYA.1.The Thiazolyl Blue Tetrazolium Blue(MTT)colorimetric assay was employed to detect proliferation of U251 cell.2.Wound healing assay and Transwell test were used to detect cell migration ability.3.Transwell chamber invasion test was employed to detect the invasion ability.4.Western blot was performed to detect U251 cell proliferation and migration related protein p38 expression.Results 1.The MTT colorimetic assay showed that surviyal rate of U251 cells treated with HYSA was lower than the blank control group and it was in a concentration dependent manner.TMZ could inhibit U251 cell proliferation as Well.Compared to TMZ alone,the inhibitory effect of the combination of TMZ and HYSA on cell proliferation was more significant(P<0.01).2.The results of Wound healing assay showed that HYSA could reduce cell invasion.After being treated with HYSA,the invasion distance was decreased.The middle and high concentration test groups had significant difference compared with the blank control group.In addition,the suppression effect of using TMZ in combination with HYSA was higher than TMZ only(p<0.05).it was suggested that there are cooperation between TMZ and HYSA in inhibiting invasion.Furthermore,the transwell assay showed exactly the same result.3.The number of cells entered the lower chamber in test groups treated with different concentration of HYSA were less than that in blank control group.The optical density(OD)value was decreased(P<0.05).4.The expression of phosphorylated p38(p-p38)was significantly reduced after treatment with HYSA.Conclusion 1.HYSA could significantly suppress the ability of proliferation,migration and invasion of human glioma U251 cell line.2.HYSA could reduce the expression of p-p38.Influencing the p38MAPK signaling pathway may be one mechanism of anti-glioma effect of HYSA.3.There was cooperation between HYSA and TMZ in anti-glioma.