| Background Glioma remains the most common cause of brain cancer-related mortality.Glioma accounts for 50-60% of brain cancer.Due to their low toxicity and infrequent side effects,traditional herbs have been increasingly popular.Coptis Chinensis is commonly used as favourite choice drug to intervent tumors with other Chinese herbs.However,little is known about its biological functions and mechanisms on intracranial gliomas.In view of this,we systematically explored the inhibitory effect of Coptis Chinensis on the growth of glioma cells,we carried out in vivo and vitro experiments.Methods vivo experiments:at first,we choosed U87 glioma cells to transplant into nude mice as models.Then randomly divided into the groups who contain drug group and control group.Gavage with processed Coptis Chinensis for more than one month at intervals of one day at doses of 20 or 10 mg/per mouse/day in drug group;Same time with distilled water in control group.Every three days,we measured the volume of the tumor.In addition,Statistics survival curves in those groups to observe the role of Cop.vitro experiments:we choosed U251 and U87 cells as samples used for vitro experiment,divided them into COP administration group and drug-free control group.Proliferation inhibition by MTT Assay for exploring the effects of COP on different types of glioma cells.A plate cloning assay to detect COP capability to affect taking shape as a globe by glioma U251 and U87 cells;Scratch assay to explore the effect of COP on inhibiting migration ability in glioma U251 and U87 cells;Flow cytometry to explore COP on cell cycles and the effect of COP on apoptosis in glioma U251 and U87 cells;Western blotting to detect the influence of HDAC3,P-STAT3 and Caspase3 after the treatment of Coptis Chinensis.Results In the transplant model with U87 glioma cells,compare with untreated group,mice in Coptis drug group can survive longer and the growth rate of their tumors is significantly slowed down.MTT assay evidenced that tumor cells can be restrained obviously in their proliferation process.The degree of action is related with COP dose and effect time.In addition,VPA also had certain inhibitory effect on the glioma cells at time-dependence;COP has low toxicity to BV2 microglial cells.Western Blot showed that 2.0 m M VPA or 5 mg/ml Coptis Chinensis could also reduce the expression of P-STAT3,and Coptis Chinensis down-regulated HDAC3,P-STAT3 and Caspase 3 in glioma cells at dose-dependent.After application with COP for glioma cells,the speed not only crawl but also migrate were decreased in evidence.As a result of COP affection,low concertration and doubled,for times lead respectively to be blocked 10.08%,16.06%,17.82% and 27.11% of the U251 cells and 11.41%,17.39%,27.32% and41.81% of the U87 cells were in G2/M phase.Additionally,COP can significantly inhibited glioma cells growth and reproduction from the clone test.Conclusion Our tests include cell and animal experiments found that 1)COP could retard the growing speed of tumors come from transplanting U87 cells to mice subcutaneous and extend their lifespan juxtapose with control group;2)COP could antagonize U87 cells which were be injected in mice body to turn into tumor shape;3)COP could restrain the capability of moving of U87 and U251 cells;4)COP not just resisted the circuit of tumor cell cycle,moreover it could induce putrescence obviously;5)COP decline STAT3 phosphorylated and decreasing HDAC3 and Caspase 3 proteins in glioma cells.To sum up,these results implicate that COP can inhibit glioma cells in multi-angle.COP down-regulated the phosphorylated STAT3 level by reducing the expression of HDAC3 protein,which affected the function of glioma cells and their biological characteristics.Multiple types of evidence illustrate that COP could restrain glioma cells from multiple mechanisms,and COP can paly the more important role in confrontation of glioma. |
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