Chinese Journal of Contemporary Neurology and Neurosurgery (Jun 2017)
Study on schizandrin A reversing drug resistance of glioma stem/progenitor cells by ATP binding cassette subfamily B member 1
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 μ mol/L group and 25.00 μ mol/L group were significantly decreased comparing with schizandrin A 0 μ mol/L group (P = 0.027, 0.006), especially in schizandrin A 25.00 μ mol/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. DOI: 10.3969/j.issn.1672-6731.2017.06.009
