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中华细胞与干细胞杂志(电子版) ›› 2018, Vol. 08 ›› Issue (05) : 292 -296. doi: 10.3877/cma.j.issn.2095-1221.2018.05.006

所属专题: 文献

论著

外泌体MicroRNA-1246促进星形胶质瘤细胞增殖与侵袭的研究
解学军1, 张冰1,()   
  1. 1. 024000 赤峰,赤峰学院附属医院神经内科
  • 收稿日期:2018-05-22 出版日期:2018-10-01
  • 通信作者: 张冰

Exosomal miRNA-1246 promotes proliferation and invasion of astroglioma cells through targeting CADM1

Xuejun Xie1, Bing Zhang1,()   

  1. 1. Department of Neurology, Affiliated Hospital of Chifeng University, Chifeng 024000, China
  • Received:2018-05-22 Published:2018-10-01
  • Corresponding author: Bing Zhang
  • About author:
    Corresponding author:Zhang Bing, Email:
引用本文:

解学军, 张冰. 外泌体MicroRNA-1246促进星形胶质瘤细胞增殖与侵袭的研究[J/OL]. 中华细胞与干细胞杂志(电子版), 2018, 08(05): 292-296.

Xuejun Xie, Bing Zhang. Exosomal miRNA-1246 promotes proliferation and invasion of astroglioma cells through targeting CADM1[J/OL]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2018, 08(05): 292-296.

目的

探讨星形胶质瘤细胞来源的外泌体中microRNA-1246(miRNA-1246)是否作用于星形胶质瘤细胞,促进其增殖与侵袭。

方法

实验分为对照组、miRNA-1246抑制剂组与miRNA-1246模拟物组,各组设6个复孔。首先从患者血液中分离外泌体并鉴定其成分。通过基质胶包被的Transwell小室实验检测星形胶质瘤细胞在miRNA-1246作用下侵袭能力的变化,CCK-8实验检测细胞增殖能力。利用荧光素酶报告基因验证miRNA-1246是否靶向细胞黏附分子1(CADM1)基因。最后通过Western Blot实验与RT-qPCR实验检测癌症组织中CADM1蛋白水平的含量并分析其与胶质瘤的关系。采用方差分析和t检验进行统计学分析。

结果

恶性胶质瘤患者血液循环外泌体中miRNA-1246的含量为2.83±1.70,高于对照组1.00±0.50,差异具有统计学意义(t?=?6.044,P?=?0.026)。转染miRNA-1246抑制剂后细胞CADM1蛋白水平为1.79±0.17,高于对照组1.00±0.09(t?=?4.576,P?=?0.017),细胞侵袭数量为(48.40±5.90)个,低于对照组96.50±6.70,而转染miRNA-1246模拟物后细胞侵袭数量为(123.20±9.80)个,高于对照组(96.50±6.70)个(t?=?5.258,P?=?0.002)。CCK-8实验中转染miRNA-1246抑制剂组A450值为0.49±0.08,低于对照组0.76±0.06,而转染miRNA-1246模拟物组A450值为1.03±0.09,显著升高(F?=?33.82,P?=?0.005)。荧光素酶报告实验表明细胞转染miR-?1246模拟物后荧光强度为4.98±1.86,低于对照组10.34±2.60(t?=?7.235,P?=?0.006),而CADM1-Mut组内之间比较差异无统计学意义。

结论

胶质瘤细胞外泌体中的miRNA-1246可通过靶向CADM1基因抑制蛋白表达,促进胶质瘤细胞的增殖与转移,提示循环外泌体中的miRNA-1246可作为恶性胶质瘤诊断与治疗的潜在靶点。

Objective

To explore whether exosomal miRNA-1246 promotes proliferation and invasion acts of astroglioma cells.

Methods

Exosomes were isolated from the patient's blood and their components were identified. The changes in the invasiveness of astroglioma cells under the action of microrna-1246 were examined by transwell chamber test. Luciferase reporter assay was used to verify whether cell adhesion molecule 1 (CADM1) is the target gene of miRNA-1246. Finally, Western Blot and RT-qPCR were used to detect CADM1 protein in cancer tissues and analyzed its relationship with glioma. ANOVA and t test were used for statistical analysis.

Results

The contents of microRNA-1246 in the exosomes of blood circulation in patients with malignant glioma (2.83±1.70) were significantly increased compared with control group (1.00±0.50) (P?<?0.05). Cell invasion in the miRNA-1246 inhibitor group (48.40±5.90) decreased compared with the control group (96.50±6.70). In the luciferase reporter gene assay, fluorescence intensity of the miRNA-1246 mimic group (4.98±1.86) significantly decreased compared with the control group (10.34±2.60) (P?<?0.05). In vitro experiments showed that microRNA-1246 can affect the expression of CADM1 gene, and thereby affect the proliferation and migration of glioma cells.

Conclusion

MicroRNA-1246 in glioma cell derived exosomes can promote the proliferation and metastasis of glioma cells by inhibiting CADM1 expression, indicating that microRNA-1246 can be used as a potential target for the diagnosis and treatment of malignant glioma.

图1 透射电镜观察外泌体(×1000)
图2 Western Blot检测表面标志蛋白CD63与HSG101
图3 外泌体miRNA-1246水平检测
图4 普通显微镜下观察胶质瘤细胞侵袭能力(结晶紫染色,×200)
表1 U-118 MG细胞转染后侵袭数量与CCK-8实验(±s
图5 质瘤组织中CADM1蛋白含量与外泌体miRNA-1246水平的关系
1
段国升. 脑胶质瘤临床治疗的进展[J]. 中华神经外科杂志, 2004, 20(2):85-87.
2
Stupp R, Mason WP, Van Den Bent MJ, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma[J]. N Engl J Med, 2005, 352(10):987-996.
3
Wang H, Wang H, Shen W, et al. Insulin-like growth factor binding protein enhances glioblastoma invasion by activating invasion- enhancing genes[J]. Cancer Res, 2003, 63(15):4315-4321.
4
李少一, 高芸, 刘宏宇, 等. 高侵袭性胶质瘤细胞转移相关miRNA的芯片分析[J]. 中国医科大学学报, 2012, 41(12):1088-1090.
5
Valadi H, Ekström K, Bossios A, et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells[J]. Nat Cell Biol, 2007, 9(6):654-659.
6
杨子楠, 魏继武. 外泌体在肿瘤发展中的研究进展[J]. 肿瘤, 2011, 31(6):565-569.
7
Sakurai-Yageta M, Masuda M, Tsuboi Y, et al. Tumor suppressor CADM1 is involved in epithelial cell structure[J]. Biochem Biophys Res Commun, 2009, 390(3):977-982.
8
刘莉, 杨永姣, 陈业刚, 等. 细胞粘附分子-1抑制膀胱癌细胞侵袭转移的机制研究[J]. 重庆医科大学学报, 2018, 43(4):510-513.
9
Chai S, Ng KY, Tong M, et al. Octamer 4/microRNA-1246 signaling axis drives Wnt/β-catenin activation in liver cancer stem cells[J]. Hepatology, 2016, 64(6):2062-2076.
10
Hummon AB, Lim SR, Difilippantonio MJ, et al. Isolation and solubilization of proteins after TRIzol extraction of RNA and DNA from patient material following prolonged storage[J]. Biotechniques, 2007, 42(4):467-470, 472.
11
王伟民, 施冲, 李天栋, 等. 脑功能区胶质瘤的手术策略[J]. 中华神经外科杂志, 2004, 20(2):71-74.
12
张玮, 彭澎, 沈铿. 外泌体来源RNA在细胞通讯中的作用[J]. 中国医学科学院学报, 2016, 38(4):480-483.
13
Lu J, Getz G, Miska EA, et al. MicroRNA expression profiles classify human cancers[J]. Nature, 2005, 435(743):834-838.
14
Wei F, Ma CY, Zhou T, et al. Exosomes derived from gemcitabine-resistant cells transfer malignant phenotypic traits via delivery of miRNA-222-3p[J]. Mol Cancer, 2017, 16(1):132.
15
Sun Z, Meng CT, Wang SH, et al. MicroRNA-1246 enhances migration and invasion through CADM1 in hepatocellular carcinoma[J]. BMC Cancer, 2014, 14(1):616.
16
Yamada N, Tsujimura N, Kumazaki M, et al. Colorectal cancer cell-derived microvesicles containing microRNA-1246 promote angiogenesis by activating Smad 1/5/8 signaling elicited by PML down-regulation in endothelial cells[J]. BBA-Gene Regulatory Mechanisms, 2014, 1839(11):1256-1272.
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