摘要
目的:探讨解毒消癌方对脑胶质瘤细胞侵袭、迁移及上皮间质转化的影响。方法:培养胶质瘤细胞株SHG44,在培养基中加入不同质量浓度(0、25、50及100μg/ml,分别为空白对照组、低剂量组、中剂量组及高剂量组)的解毒消癌方水提物,采用细胞划痕实验检测其对迁移能力的影响,采用Transwell侵袭实验检测其对细胞侵袭能力的影响。使用胶质瘤细胞构建裸鼠移植瘤,成瘤后随机分为空白对照组、低剂量组、中剂量组和高剂量组,分别给予0、25、50及100μg/ml解毒消癌方喂养,采用蛋白质印迹法检测裸鼠移植瘤内ZEB1、snail、转化生长因子β(TGF-β)、基质金属蛋白酶9(MMP-9)蛋白表达水平。结果:细胞划痕实验结果显示,经解毒消癌方处理后,低、中及高剂量组肿瘤细胞迁移距离明显短于空白对照组,差异均有统计学意义(P<0.05),细胞迁移能力受到明显抑制;且解毒消癌方对胶质瘤细胞迁移能力的抑制呈时间和浓度依赖性,时间越长、浓度越高,细胞迁移能力越弱。Transwell侵袭实验结果显示,经解毒消癌方处理后,低、中及高剂量组侵袭细胞数明显低于空白对照组,差异均有统计学意义(P<0.05),胶质瘤细胞侵袭能力受到明显抑制;且解毒消癌方对胶质瘤细胞侵袭能力的抑制呈浓度依赖性,浓度越高,细胞侵袭能力越弱。蛋白质印迹法检测结果显示,经解毒消癌方喂养后,大鼠胶质瘤细胞TGF-β、ZEB-1、snail及MMP-9蛋白表达明显降低,低、中及高剂量组均低于空白对照组,差异均有统计学意义(P<0.05);且解毒消癌方对TGF-β、ZEB-1、snail及MMP-9蛋白表达的抑制呈浓度依赖性,浓度越高,表达越低。结论:解毒消癌方能够阻止胶质瘤细胞上皮间质转化,进而抑制细胞侵袭、迁移,其作用机制与下调细胞中TGF-β、ZEB1、snail及MMP-9等多种效应蛋白表达有关。
OBJECTIVE:To probe into the effects of Jiedu Xiao’ai formula on invasion,migration and epithelial mesenchymal transformation of glioma cells.METHODS:The glioma cell line SHG44 was cultured,and different concentrations(0,25,50,100 μg/ml,respectively,as blank control group,low-dose group,medium dose group and high-dose group) of the water extract of Jiedu Xiao ’ai formula were added to the medium.The cell scratch test was used to detect its influence on the migration ability,and Transwell invasion test was used to detect its influence on the cell invasion ability.Glioma cells were used to construct transplanted tumor in nude mice.After tumor formation,the mice were randomly divided into blank control group,low dose group,medium dose group and high-dose group,and fed with 0,25,50 and 100 μg/ml Jiedu Xiao ’ai formula,respectively.Western blot was used to detect the protein expression levels of ZEB1,snail,transforming growth factor β(TGF-β) and matrix metalloproteinase 9(MMP-9) in transplanted tumors of nude mice.RESULTS:Cell scratch test showed that the migration distance of tumor cells in the low,medium and high dose group was significantly shorter than that in the blank control group after treatment with Jiedu Xiao’ai formula(P<0.05),and the migration ability of cells was significantly inhibited.The inhibition of Jiedu Xiao’ai formula on glioma cell migration ability was time and concentration dependent.The longer the time and the higher the concentration,the weaker the cell migration ability.Transwell invasion experiment showed that the number of tumor cells invaded in the low,medium and high dose groups was significantly lower than that in the blank control group after being treated with Jiedu Xiao ’ai formula,the difference was statistically significant(P<0.05),the cell invasion ability was significantly inhibited,and the inhibition of the glioma cell invasion ability of Jiedu Xiao ’ai formula was concentration dependent.The higher the concentration,the weaker the cell invasion ability.Western blot showed that the expressions of TGF-β,ZEB-1,snail and MMP-9 protein in glioma cells decreased significantly after being fed with Jiedu Xiao ’ai formula,and the low,medium and high dose groups were all lower than the blank control group,the differences were statistically significant(P<0.05).The inhibition of the expression of TGF-β,ZEB-1,snail and MMP-9 by Jiedu Xiao’ai formula was concentration dependent.The higher the concentration,the lower the expression.CONCLUSIONS:Jiedu Xiao’ai formula can prevent the epithelial mesenchymal transformation of glioma cells,thereby inhibiting cell invasion and migration.Its mechanism of action is related to down-regulating the expression of various effector proteins such as TGF-β,ZEB1,snail and MMP-9 in cells.
作者
高爽
GAO Shuang(Dept.of Neurosurgery,Tumor Hospital Affiliated to Harbin Medical University,Heilongjiang Harbin 150040,China)
出处
《中国医院用药评价与分析》
2020年第12期1424-1428,1432,共6页
Evaluation and Analysis of Drug-use in Hospitals of China
基金
黑龙江省中医药科研项目(No.ZHY19-W0412)。
关键词
解毒消癌方
胶质瘤
裸鼠
细胞迁移
细胞侵袭
上皮间质转化
Jiedu Xiao ai formula
Glioma
Nude mice
Cell migration
Cell invasion
Epithelial mesenchymal transformation
