[1]陶　祥　陈　晨　陶　芸　张申起　刘宝辉　廖健明　张文斐　陈谦学.SNAI2与E-cadherin参与LRIG1对U251细胞侵袭与转移的抑制作用[J].中国临床神经外科杂志,2016,(09):536-539.[doi:10.13798/j.issn.1009-153X.2016.09.009]
　TAO Xiang,CHEN Chen,TAO Yun,et al.Role of SNAI2 and E-cadherin in regulation of invasiveness and metastasis of human gliomas U251 cells by LRIG1[J].,2016,(09):536-539.[doi:10.13798/j.issn.1009-153X.2016.09.009]
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SNAI2与E-cadherin参与LRIG1对U251细胞侵袭与转移的抑制作用()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
期数:
2016年09期
页码:
536-539
栏目:
论著
出版日期:
2016-09-25

文章信息/Info

Title:
Role of SNAI2 and E-cadherin in regulation of invasiveness and metastasis of human gliomas U251 cells by LRIG1
文章编号:
1009-153X（2016）09-0536-04
作者:
陶　祥　陈　晨　陶　芸　张申起　刘宝辉　廖健明　张文斐　陈谦学
430060 武汉，武汉大学人民医院神经外科（陶　祥、张申起、刘宝辉、廖健明、张文斐、陈谦学）；250012 济南，山东大学口腔医学院（陈　晨）；442000 十堰，湖北医药学院口腔医学系（陶　芸）
Author(s):
TAO Xiang1;  CHEN Chen2;  TAO Yun3;  ZHANG Shen-qi1;  LIU Bao-hui1;  LIAO Jian-ming1;  ZHANG Wen-fei1;  CHEN Qian-xue1.
1.Department of Neurosurgery, Renmin Hospital, Wuhan University, Wuhan 430060, China;
2. School of Stomatology, Shandong University, Ji'an 250012,China;
3. Department of Stomatology, Hubei University of Medicine, Shiyan 442000, China
关键词:
胶质瘤; U251细胞; LRIG1; SNAI2; E-cadherin; 侵袭; 转移
Keywords:
Glioma U251 cells;  LRIG1;  SNAI2;  E-cadherin;  Invasiveness;  Migration
分类号:
R 739.41; Q 786
DOI:
10.13798/j.issn.1009-153X.2016.09.009
文献标志码:
A
摘要:
目的 探讨多亮氨酸重复区免疫球蛋白样蛋白1（LRIG1）对人脑胶质瘤U251细胞侵袭与转移及U251细胞SNAI2、E-cadherin表达的影响。方法 传代培养U251细胞，随机分为空白对照组、LRIG1空载体组、LRIG1过表达组、LRIG1低表达组、LRIG1低表达阴性对照组，应用脂质体介导的基因转染技术分别转染PBS、PEGFP-N1-U251质粒、PEGFP-LRIG1-U251质粒、LRIG1-siRNA、LRIG1-NC-siRNA。应用PCR、Western blot检测细胞LRIG1、SNAI2、E-cadherin mRNA和蛋白表达水平，Transwell侵袭实验检测细胞侵袭能力，划痕实验检测细胞转移能力。结果 LRIG1过表达组U251 SNAI2表达下调（P<0.05），E-cadherin表达上调（P<0.05），细胞侵袭与转移能力明显下降（P<0.05）。LRIG1低表达组U251细胞SNAI2表达上调（P<0.05），E-cadherin表达下调（P<0.05），细胞侵袭与转移能力明显提高（P<0.05）。结论 上调LRIG1可抑制U251细胞侵袭与转移，而敲除LRIG1可明显促进U251细胞侵袭与转移，SNAI2及E-cadherin可能参与其调节过程。
Abstract:
Objective To investigate the role of leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) in regulation of invasiveness and metastasis of human glioma U251 cells and the effects of SNAI2 and E-cadherin on it. Methods The cultured human glioma U251 cells were randomly divided into five groups, i.e. blank control group, LRIG1 vector group, LRIG1 over-expression group, LRIG1 low-expression group and LRIG1 low-expression negtive control group, in which the PBS, PEGFP-N1 plasmid,PEGFP-LRIG1 plasmid, LRIG1-siRNA and LRIG1-NC-siRNA wrer transfecte into the U251 cells, respectively. The mRNA and protein expressions of LRIG1, SNAI2 and E-cadherin were detected by real time-PCR and Western blotting, respectively. The invasiveness of U251 cells was determined by transwell chamber assay, and the migration of U251 cells was detected by transwell chamber assay with a matrigel coating and wound healing test. Results The levels of LRIG1 and E-Cadherin mRNA and proteins expressions were significantly higher in LRIG1 over-expression group than other groups (P<0.01), and they were significantly lower in LRIG1 low-expression group than blank control, LRIG1 vector group and low-expression negtive control groups (P<0.05). The levels of SNAI2 mRNA and protein expressions were significantly higher in LRIG1 low-expression group than other groups (P<0.05), and they were significantly lower in LRIG1 over-expression group than blank control, LRIG1 vector group and low-expression negtive control groups (P<0.01). The invasive and migration activities of U251 were significantly inhibited in LRIG1 over-expression group compared to other groups (P<0.01), and they were significantly enhanced in LRIG1 low-expression group compared to blank control, LRIG1 vector group and low-expression negtive control groups (P<0.01). Conclusion It is suggested that LRIG1 regulates the invasion and migration of gliomas U251 cells probably via the activation of SNAI2 and E-cadherin signaling pathway.

参考文献/References:

[1]Mao F, Wang B, Xiao Q, et al. A role for LRIG1 in the regu- lation of malignant glioma aggressiveness [J]. Int J Oncol, 2013, 42(3): 1081-1087.
[2] 郭振涛，刘宝辉，邓　钢，等. LRIG1抑制人脑胶质瘤细胞 增殖的研究[J]. 中国临床神经外科杂志，2013，18：606- 609.
[3] 丁　昊，周志伟，易　伟，等. LRIG1相关功能片段对 EGFR活性和胶质瘤细胞增殖的影响[J]. 中国临床神经 外科杂志，2015，20：544-546，575.
[4] Stutz MA, Shattuck DL, Laederich MB, et al. LRIG1 nega- tively regulates the oncogenic EGF receptor mutant EGFRvIII [J]. Oncogene, 2008, 27(43): 5741-5752.
[5] Yang HW, Menon LG, Black PM, et al. SNAI2/Slug pro- motes growth and invasion in human gliomas [J]. BMC Cancer, 2010, 10: 301.
[6] Barami K, Lewis-Tuffin L, Anastasiadis PZ. The role of cadherins and catenins in gliomagenesis [J]. Neurosurg Focus, 2006, 21(4): E13.
[7] Yu H, Shen Y, Hong J, et al. The contribution of TGF-β in Epithelial-Mesenchymal Transition (EMT): Down-regula- tion of E-cadherin via snail [J]. Neoplasma, 2015, 62(1): 1- 15.
[8] Ye F, Gao Q, Xu T, Zeng L, et al. Upregulation of LRIG1 suppresses malignant glioma cell growth by attenuating EGFR activity [J]. Neurooncol, 2009, 94(2): 183-194.
[9] Zhong D, He G, Zhao S, et al. LRG1 modulates invasion and migration of glioma cell lines through TGF-β signaling pathway [J]. Acta Histochem, 2015, 117(6): 551-558.
[10] Sheu JJ, Lee CC, Hua CH, et al. LRIG1 modulates aggres- siveness of head and neck cancers by regulating EGFR- MAPK-SPHK1 signaling and extracellular matrix remo- deling [J]. Oncogene, 2014, 33 (11): 1375-1384.
[11] Xia H, Cheung WK, Ng SS, et al. Loss of brain-enriched miR-124 microRNA enhances stem-like traits and invasi- veness of glioma cells [J]. Biol Chem, 2012, 287(13): 9962- 9971.
[12] Guichet PO, Guelfi S, Teigell M, et al. Notch1 stimulation induces a vascularization switch with pericyte-like cell differentiation of glioblastoma stem cells [J]. Stem Cells, 2015, 33 (1): 21-34.
[13] Zhang X, Song Q, Wei C, et al. LRIG1 inhibits hypoxia- induced vasculogenic mimicry formation via suppression of the EGFR/PI3K/AKT pathway and epithelial to mesenchy- mal transition in human glioma SHG-44 cells [J]. Cell Stress Chaperones, 2015, 20 (4): 631-641.

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备注/Memo

备注/Memo:
基金项目：国家自然科学基金（81372683）
通讯作者：陈谦学，E-mail：chenqx666@sohu.com
更新日期/Last Update: 2016-09-30