新型多金属氧酸盐化合物{BiW_(8)O_(30)}对胶质母细胞瘤细胞体内、体外抑制作用的研究

Inhibitory effects of a novel polyoxometalate compound{BiW_(8)O_(30)}on glioblastoma cells in vivo and in vitro

目的:明确新型多金属氧酸盐化合物{BiW_(8)O_(30)}对胶质母细胞瘤细胞体内、体外抑制作用的影响,并探讨可能的作用机制。方法:采用不同浓度{BiW_(8)O_(30)}处理LN229、U251细胞。CCK-8法检测细胞增殖能力;细胞划痕和侵袭实验检测细胞迁移和侵袭能力;流式细胞实验检测细胞凋亡情况;DAPI染色观察细胞核形态变化;裸鼠皮下移植瘤模型实验检测肿瘤体内增殖能力;Western blot实验检测细胞中凋亡相关蛋白的表达水平;蛋白质组测序技术分析{BiW_(8)O_(30)}发挥对胶质母细胞瘤细胞抑制作用的可能机制。结果:与对照组相比,{BiW_(8)O_(30)}处理组的细胞存活率、划痕愈合率、侵袭细胞数均显著降低(P<0.001)。流式细胞实验显示,{BiW_(8)O_(30)}处理提高肿瘤细胞凋亡率(P<0.05)。DAPI染色结果显示,{BiW_(8)O_(30)}处理后,肿瘤细胞出现核浓缩、核碎裂等形态变化。{BiW_(8)O_(30)}处理显著抑制异种移植肿瘤在裸鼠体内的增殖能力(P<0.05)。Western blot结果显示,与对照组相比,{BiW_(8)O_(30)}处理组凋亡相关蛋白XIAP表达降低(P<0.001),caspase 3表达升高(P<0.001)。蛋白质组测序分析显示,{BiW_(8)O_(30)}处理后胶质母细胞瘤细胞内共有559个蛋白的表达出现显著差异,其中250个蛋白上调,309个蛋白下调(符合adj.P-value<0.05,|Fold change|>1.5)。通过基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,发现差异蛋白主要富集于程序性细胞死亡的调控、细胞凋亡过程、细胞运动的调节、细胞黏附等通路。结论:{BiW_(8)O_(30)}抑制胶质母细胞瘤细胞体内、体外的增殖能力,体外迁移和侵袭能力,并可诱导肿瘤细胞凋亡,其机制可能与细胞凋亡过程、细胞黏附与运动等通路相关。

Objective:To investigate the inhibitory effect of a novel polyoxometalate compound{BiW_(8)O_(30)}on glioblastoma LN229 and U251 cells in vivo and in vitro,and the possible mechanism.Methods:LN229 and U251 cells were treated with different concentrations of{BiW_(8)O_(30)}.CCK-8 assay was used to detect cell proliferation.Cell scratch and invasion assay to detect cell migration and invasion.Cell apoptosis was detected by flow cytometry.Nuclear morphological changes were observed by DAPI staining.The proliferation ability of tumor in vivo was detected by subcutaneous transplantation of tumor model in nude mice.Western blot was used to detect the expression level of apoptosis-related proteins in cells.Proteomic sequencing was used to analyze thepossible mechanisms of inhibitory effect of{BiW_(8)O_(30)}on glioblastoma cells.Results:Compared with the control group,the cell survival rate,scratch healing rate and invasive cell number of{BiW_(8)O_(30)}treatment group were significantly decreased(P<0.001).The apoptosis rate of the{BiW_(8)O_(30)}treatment group was significantly higher than that of the control group(P<0.05).DAPI staining results showed that some cells in the{BiW_(8)O_(30)}group showed morphological changes such as nuclear condensation and nuclear fragmentation.{BiW_(8)O_(30)}inhibited the growth of LN229 xenografts in nude mice compared with control group(P<0.05).Western blot results showed that compared with the control group,the expression of apoptosis-related protein XIAP was decreased and caspase 3 was increased in the{BiW_(8)O_(30)}group(P<0.05).Proteome sequencing analysis showed that a total of 559 proteins were significantly differentially expressed before and after{BiW_(8)O_(30)}treatment,of which 250 proteins were up-regulated and 309 proteins were down-regulated(consistent with adj.P-value<0.05,|Fold change|>1.5).Through GO and KEGG enrichment analysis,it was found that differential proteins were enriched in pathways such as regulation of programmed cell death,apoptotic process,regulation of cell motility,cell adhesion.Conclusion:{BiW_(8)O_(30)}inhibits the proliferation in vivo and in vitro,migration and invasion of glioblastoma cells,and induces cell apoptosis.The possible mechanisms may be related to pathways of apoptosis process,cell adhesion and motility.