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[1]李启凤,李一凡,张金芳,等.没食子酸甲酯抑制胶质瘤细胞中Akt 和ERK1/2 活性发挥抗肿瘤作用[J].宁夏医科大学学报,2020,(08):757-764.[doi:10.16050/j.cnki.issn1674-6309.2020.08.001]
 LI Qifeng,LI Yifan,ZHANG Jinfang,et al.Methyl Gallate Exerts Anti-tumor Effects by Inhibiting Akt and ERK1/2 Activity in Glioma Cells[J].Ningxia Medical University,2020,(08):757-764.[doi:10.16050/j.cnki.issn1674-6309.2020.08.001]
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没食子酸甲酯抑制胶质瘤细胞中Akt 和ERK1/2 活性发挥抗肿瘤作用(PDF)
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2020年08期
页码:
757-764
栏目:
论著
出版日期:
2021-04-17

文章信息/Info

Title:
Methyl Gallate Exerts Anti-tumor Effects by Inhibiting Akt and ERK1/2 Activity in Glioma Cells
文章编号:
1674-6309(2020)08-0757-08
作者:
李启凤1李一凡2张金芳1王开昕1
1. 华中科技大学协和深圳医院(深圳市南山区人民医院)病理科,深圳 518000; 2. 华中科技大学协和深圳医院(深圳市南山区人民医院)中心实验室,深圳 518000
Author(s):
LI Qifeng1 LI Yifan2 ZHANG Jinfang1 WANG Kaixin1
1. Department of Pathology, Concord Shenzhen Hospital of Huazhong University of Science and Technology (Shenzhen Nanshan People’s Hospital),Shenzhen 518000,China; 2. Central Laboratory of Concord Shenzhen Hospital of Huazhong University of Science and Technology (Shenzhen Nanshan People’s Hospital), Shenzhen 518000,China
关键词:
没食子酸甲酯胶质瘤细胞迁移AktERK1/2
Keywords:
methyl gallate glioma cells migration Akt ERK1/2
分类号:
R285.5
DOI:
10.16050/j.cnki.issn1674-6309.2020.08.001
文献标志码:
A
摘要:
目的 探讨没食子酸甲酯(methyl gallate,MG)对大鼠胶质瘤细胞C6和人胶质瘤细胞 U373增殖和迁移的影响以及抗肿瘤活性和调控机制。方法 MG从簇毛槭茎皮中分离提取,通过MTT法检测MG对胶质瘤细胞活性的影响,BrdU法检测胶质瘤细胞的增殖,应用划痕实验分析MG对胶质瘤细胞迁移的影响。Western blot检测MG对Akt和ERK1/2活性的影响。免疫荧光染色实验观察黏着斑的形成。结果 细胞划痕试验显示,对照组划痕基本愈合,MG(5 μg·mL-1)处理组划痕愈合被抑制(P<0.05); 外源性Glu可增加细胞的迁移速度和距离,MG预处理抑制Glu诱导的细胞活性和迁移(P均<0.05)。Western blot结果显示,在向C6和U373细胞中加入Glu(终浓度150 μM)后,Akt的磷酸化水平无变化(P>0.05),ERK1/2磷酸化水平升高(P<0.05)。免疫荧光染色结果显示,对照组和Glu处理组中,鬼笔环肽染色无变化,FA的数量和规模增加(P<0.05)。NBQX可抑制细胞的存活和迁移 (P<0.05)。终浓度50 μM的钙离子螯合剂BAPTA-AM预处理后,Akt磷酸化水平降低,细胞死亡增加(P<0.05)。用5 μg·mL-1MG处理细胞,20 min后加入Glu刺激,Western blot 结果表明,MG不仅抑制了Akt的磷酸化水平,还抑制了Glu诱导ERK1/2磷酸化;MG抑制Glu诱导的桩蛋白Ser 83磷酸化(P<0.05)。结论 MG可抑制大鼠胶质瘤细胞C6和人胶质瘤细胞 U373增殖和迁移,可能与抑制Akt和ERK1/2信号通路有关。
Abstract:
Objective To investigate the effects of methyl gallate(MG)on the proliferation and migration of rat glioma cells C6 and human glioma cells U373,and to study its anti-tumor activities and regulatory mechanisms. Methods MG was isolated and extracted from the stem bark of Acer barbinerve. The viability, proliferation and migration of glioma cells treated with MG were analyzed using MTT method, BrdU method and scratch test, respectively. The effects of MG on Akt and ERK1/2 activities were detected by Western blot, and the formation of focal adhesion(FA) was detected by immunofluorescence staining. Results Cell scratch test showed that the scratch healing in the control group was basically healed,while the scratch healing in the MG(5 μg· mL-1) treatment group was significantly inhibited(P<0.05). Exogenous Glu significantly increased the cell migration rate and distance, while MG pretreatment significantly inhibited the cell proliferation induced by Glu ,activity and migration (P all<0.05). Western blot results showed that after adding Glu (final concentration 150 meters) to C6 and U373 cells, there was no significant change in the phosphorylation level of Akt(P>0.05), but the phosphorylation level of ERK1/2 was significantly increased(P<0.05). Immunofluorescence staining results showed that there was no significant difference between the control group and the Glu treatment group, but the number and size of FA increased significantly(P<0.05). NBQX significantly inhibited cell survival and migration (P<0.05). After pretreatment with bapta-am,a calcium ion chelating agent with a final concentration of 50 meters, Akt phosphorylation level was significantly reduced and cell death was significantly increased(P<0.05).After 20 min,Glu stimulation was added. Western blot results showed that MG not only inhibited the phosphorylation level of Akt, but also inhibited the phosphorylation of ERK1/2 induced by Glu.MG inhibits Ser83 phosphorylation induced by Glu(P<0.05). Conclusion MG can inhibit the proliferation and migration of rat glioma cell C6 and human glioma cell U373,which may be related to the inhibition of Akt and ERK1/2 signaling pathway.

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备注/Memo

备注/Memo:
收稿日期:2019-05-29
基金项目:广东省自然科学基金自由申请项目(2017A030313170)
作者简介:李启凤(1985—),女,硕士,主治医生。研究方向:肿瘤病理。
更新日期/Last Update: 2020-08-20