Study on schizandrin A reversing drug resistance of glioma stem/progenitor cells by ATP binding cassette subfamily B member 1

WEI Zi-long; DI Chong; LOU Mei-qing; ZHAO Yao-dong

doi:10.3969/j.issn.1672-6731.2017.06.009

Abstract：Objective To investigate the effect of schizandrin A on drug resistance of glioma stem/ progenitor cells (GSPCs) and its mechanism.Methods Isolate and culture SHG-44s cells from human glioma cell line SHG-44.The SHG-44s cells were treated with different concentrations of schizandrin A (0,12.50,25.00 and 50.00 μmol/L) and vincristine (400,800 and 1200 nmol/L).The cell proliferative activity was measured by cell counting kit-8 (CCK-8) assay.Rhodamine 123 staining was used to detect the drug delivery ability of SHG-44s cells.The transcription and translation ability of ATP binding cassette subfamily B member 1 (ABCB1) gene of SHG-44s cells was detected by real-time polymerase chain reaction (PCR) and Western blotting.Results The proliferative activity of SHG-44s cells was inhibited when the concentration of schizandrin A was 50 μmol/L (P =0.001,0.001,0.039),so this concentration was removed in the follow-up study.No matter the concentration of vincristine was 400,800 or 1200 nmol/L,combining with schizandrin A could inhibit the proliferative activity of SHG-44s cells (vincristine 400 nmol/L:P =0.007,0.001;vincristine 800 nmol/L:P =0.001,0.000;vincristine 1200 nmol/L:P =0.000,0.000).Inverted fluorescence microscopy findings showed SHG-44s cells in the group of schizandrin A 0 μ mol/L rarely revealed green fluorescence,while SHG-44s cells in the groups of schizandrin A 12.50 and 25.00 μmol/L presented obvious green fluorescence.Flow cytometry showed that with the increasing of schizandrin A concentration,the percentage of positive cells by Rhodamine 123 staining was 10.40％,39.20％ and 45.20％,respectively.Real-time PCR showed that ABCB1 gene expression levels of SHG-44s cells in schizandrin A 12.50 μmol/L group and 25.00 μ mol/L group were significantly decreased comparing with schizandrin A 0 μmol/L group (P =0.027,0.006),especially in schizandrin A 25.00 μmol/L group (P =0.034).Western blotting showed that the expression level of P-glycoprotein (P-gp) in SHG-44s cells was gradually decreased with the increasing of schizandrin A concentration.Conclusions Schizandrin A can inhibit the drug delivery ability of P-gp coded by ABCB1 gene existing in the surface of GSPCs.It can further reverse the drug resistance of GSPCs by reducing the transcription and translation of ABCB1 gene.