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AIM:To study the effect of centromere protein W ( CENP-W) down-regulation on human glioma U87 cells.METHODS:Small interfering RNA ( siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot .The proliferation of the cells was analyzed by MTT assay , BrdU staining and colony formation experiment .Transwell chamber assay was used to detect the invasion a-bility of the cells .The cell migration ability was measured by a scratch test .The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry .RESULTS:The results of MTT assay , colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group .The results of Transwell and scratch experiments showed that the migra-tion and invasion abilities in experimental group were weaker than those in blank control group and negative control group . The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G 0/G1 phase .The percentage of apoptotic cells in experimental group was higher than that in control group ( P<0.05 ) .CON-CLUSION:Down-regulation of CENP-W expression inhibits the proliferation , migration and invasion of human glioma cells and promotes the apoptosis of the cells , suggesting that CENP-W may be a potential target of gene therapy for human glioma.
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Objective To study the effect of exsomes in SH-SY5Y cells after hypoxic ischemia/reperfusion injury.Methods Human umbilical vein endothelial cell hypoxic ischemia/reperfusion (HUVEC I/R) injury models were established,and the exosomes derived from HUVEC I/R were extracted and identified.SH-SY5Y cell hypoxic ischemia/reperfusion injury models (SH-SY5Y I/R) were established,and cells from SH-SY5Y I/R were divided into control group and exosomes-treatedgroup.The proliferation of SH-SY5Y cells was evaluated by CCK-8 assay 24,48and 72 h after cell inoculation.Transwell assay and wound-healing assay were used to examine the invasion and migration.Hochest33258 staining and Flow cytometry were used to monitor the changes of cell cycle and apoptosis.Expressions of Caspase-3,Bax and Bcl-2 were measured by real-time fluorescence quantificative-PCR and Western blotting.Results As compared with those in the control group,the proliferation abilities of SH-SY5Y cells in exosomes-treated group were significantly promoted (48 h:0.70±0.05 vs.0.94±0.08;72 h:0.83±0.05 vs.1.02±0.06),the cell cycle rate of S phase was significantly increased (14.39％±4.11％ vs.20.54％±3.46％),and G0/G1 phase was statistically decreased (71.26％± 5.24％ vs.66.87％±4.23％,P＜0.05).What's more,cell invasive was significantly promoted (44.00±6.56 vs.70.67±6.11),and relative wound injury area was significantly reduced in the exosomes treated group (0.61±0.07 vs.0.52±0.10);significant differences were noted between the two groups (P＜0.05).The mRNA expressions of Bax and Caspase-3 were significantly decreased and the mRNA expressions of Bcl-2 was significantly increased in the exosomes-treated group as compared with those in the control group (P＜0.05).Conclusion HUVEC I/R-derived exosomes play neuro-protective role in human SH-SY5Y cells after hypoxic I/R injury.