| Objective: To investigate the effect of all-trans retinoic acid(ATRA)on glioma cells and the correlation between Pin1 and Notch1 signalling in glioma.Methods:1.The effect of ATRA on glioma cell lines U251 and U87 proliferation was detected by CCK-8 assay.2.The expression of GFAP in U251 and U87 cells which treated with ATRA were detected by immunofluorescence staining.3.The expression of Pin1,Notch1,Hes1,MDR1 and MRP1 in U251 and U87 cells which treated with ATRA were assayed by Western blotting.4.The expression of Pin1,Notch1 and Hes1 in U251 and U87 cells treated with ATRA were identified by immunofluorescence.5.The cells(U251-pin1 and U87-Pin1)over-expressed Pin1 were obtained by lipofectamine-mediated transfection.The expression of Notch1 and Hes1 in U251-pin1 and U87-Pin1 cells were assayed by Western blotting.Results 1.The results of CCK-8 showed that ATRA could inhibit the proliferation of U251 and U87 cells,and this inhibitory effect was significantly time-dose-dependent.2.The expression of GFAP was higher in ATRA treated cells than that in the control cells.This suggested ATRA could induce differentiation of U251 and U87 cells.3.The results of Western blotting showed that the expression of Pin1,Notch1,Hes1,MDR1 and MRP1 in U251 and U87 cells were down-regulated following treatment with ATRA.4.Immunofluorescence showed the expression of Pin1,Notch1 and Hes1 in U251 and U87 cells were lower following treated with ATRA group in U251 and U87 cells.5.Western blotting showed that the expression of Pin1 was significantly increased in U251-pin1 and U87-pin1 cells,and the expression of Notch1 and Hes1 were also up-regulated.Conclusions:1.ATRA could induce differentiation and inhibit proliferation of human glioma U251 and U87 cell lines.2.ATRA downregulated the expression of Notch1 and Hes1 by inhibiting Pin1.Moreover,ATRA also inhibited expression of multi-drug resistant protein MDR1 and MRP1 in glioma U251 and U87 cells. |
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