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J Zhejiang Univ (Med Sci)  2018, Vol. 47 Issue (2): 181-186    DOI: 10.3785/j.issn.1008-9292.2018.04.12
    
Isoliquiritigenin can inhibit migration and invasion of human glioma stem cells by down-regulating matrix metalloproteinases
DANG Ying1,2(),LIN Yuliang1,SUN Hongjun3,SUN Jianjun1,LI Changdong1,LI Zhiyun1,*()
1. Department of Neurosurgery, PLA Lanzhou General Hospital, Lanzhou 730050, China
2. Department of Functional Neurosurgery, Xi'an Honghui Hospital, Xi'an 710054, China
3. Department of Neurosurgery, Second Gansu Provincial People's Hospital, Lanzhou 730030, China
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Abstract  

Objective: To investigate the effects of isoliquiritigenin on the migration and invasion of human glioma stem cells and the underlying mechanism. Methods: The stem cell markers CD133 and Nestin in SHG44 human glioma stem cells were examined with immunofluorescence microscopy. The migration and invasion ability of glioma stem cells was determined by transwell method. The mRNA and protein expression of matrix metalloproteinase (MMP)-2 and MMP-9 were detected by real-time RT-PCR and Western blot, respectively. Results: CD133 and Nestin were positive in SHG44 cells. The number of migrated cells in SHG44 cells treated with 20 and 80 μmol/L isoliquiritigenin for 48 h were significantly lower than that in control group (76±5 and 42±4 vs. 85±6, all P < 0.01), and the number of migrated cells in 80 μmol/L isoliquiritigenin group was lower than that in 20 μmol/L isoliquiritigenin group (P < 0.01). The numbers of cells crossing through membrane in 20 and 80 μmol/L isoliquiritigenin groups were 190±13 and 130±9, respectively, which were significantly lower than that in control group (230±14, all P < 0.01), and the number of crossed cells in the 80 μmol/L isoliquiritigenin group was lower than that in 20 μmol/L isoliquiritigenin group (P < 0.01). The mRNA and protein expression levels of MMP-2 and MMP-9 were decreased compared with control group (P < 0.05 or P < 0.01), and the expression levels in 80 μmol/L isoliquiritigenin group were lower than those in 20 μmol/L isoliquiritigenin group (P < 0.05 or P < 0.01). Conclusion: Isoliquiritigenin exhibits antitumor effects on glioma stem cells by inhibiting cell migration and invasion, which may be related to the down-regulation of MMP-2 and MMP-9.



Key wordsGlioma/drug therapy      Stem cells      Isoliquiritigenin      Matrix metalloproteinase 9      Matrix metalloproteinase 2      Neoplasm invasiveness      Cell movement     
Received: 01 September 2017      Published: 24 July 2018
CLC:  R739.41  
Corresponding Authors: LI Zhiyun     E-mail: dangy052@qq.com;lizhiyun456@163.com
Cite this article:

DANG Ying,LIN Yuliang,SUN Hongjun,SUN Jianjun,LI Changdong,LI Zhiyun. Isoliquiritigenin can inhibit migration and invasion of human glioma stem cells by down-regulating matrix metalloproteinases. J Zhejiang Univ (Med Sci), 2018, 47(2): 181-186.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2018.04.12     OR     http://www.zjujournals.com/med/Y2018/V47/I2/181


异甘草素通过下调基质金属蛋白酶抑制人脑胶质瘤干细胞迁移和侵袭

目的: 探讨异甘草素对人脑胶质瘤干细胞迁移和侵袭能力的影响及相关机制。方法: 免疫荧光法检测SHG44人脑胶质瘤干细胞的干性特征;transwell实验观察SHG44人脑胶质瘤干细胞的迁移和侵袭能力;实时定量RT-PCR法和蛋白质印迹法分别检测SHG44人脑胶质瘤干细胞中基质金属蛋白酶(MMP)2和MMP-9的mRNA和蛋白表达量。结果: SHG44细胞中干细胞标志物CD133和Nestin呈阳性表达。20 μmol/L和80 μmol/L的异甘草素作用于胶质瘤干细胞48 h后,迁移细胞数分别为76±5和42±4,与阴性对照组(85±6)差异有统计学意义(均P < 0.01),且80 μmol/L异甘草素组迁移细胞数少于20 μmol/L异甘草素组(P < 0.01);穿透滤膜至小室背面的细胞数分别为190±13和130±9,与阴性对照组(230±14)差异有统计学意义(均P < 0.01),且80 μmol/L异甘草素组穿透滤膜至小室背面的细胞数少于20 μmol/L异甘草素组(P < 0.01);MMP-2、MMP-9 mRNA和蛋白表达量均较阴性对照组下调(P < 0.05或P < 0.01),且80 μmol/L异甘草素组MMP-2、MMP-9 mRNA和蛋白表达量低于20 μmol/L异甘草素组(P < 0.05或P < 0.01)。结论: 异甘草素可通过下调MMP-2和MMP-9的表达抑制胶质瘤干细胞迁移和侵袭。


关键词: 神经胶质瘤/药物疗法,  干细胞,  异甘草素,  基质金属蛋白酶9,  基质金属蛋白酶2,  肿瘤侵润,  细胞运动 
基因 引物序列(5′→3′)
MMP-2 正向:GAGTTGGCAGTGCAATACCT
反向:CCAAAGAACTTCTGCATCTTCT
MMP-9 正向:TCCAAGGCCAATCCTACT
反向:CGTCGAGTCAGCTCGGGT
GAPDH 正向:TGGGTGTGAACCACGAGAA
反向:GGCATGGACTGTGGTCATGA
Tab 1 Primer sequences for real-time RT-PCR
Fig 1 Expression of specific markers CD133 and Nestin in SHG44 human glioma stem cell spheres under the fluorescence microscope
Fig 2 Effect of isoliquiritigenin on migration of SHG44 human glioma stem cells
Fig 3 Effect of isoliquiritigenin on invasion ability of SHG44 human glioma stem cells
Fig 4 mRNA expression of MMP-2 and MMP-9 genes in SHG44 human glioma stem cells treated with DMSO(negative control) and isoliquiritigenin
Fig 5 Protein expression of MMP-2 and MMP-9 in SHG44 human glioma stem cells treated with DMSO(negative control) and isoliquiritigenin
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