MiR-15b and miR-152 reduce glioma cell invasion and angiogenesis via NRP-2 and MMP-3.
MiR-15b and miR-152 reduce glioma cell invasion and angiogenesis via NRP-2 and MMP-3.
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DOI：
10.1016/j.canlet.2012.10.026
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发表时间：
2013-02-28
期刊：
影响因子：
9.7
通讯作者：
Jiang, Feng
中科院分区：
文献类型：
--
作者：
Zheng, Xuguang;Chopp, Michael;Lu, Yong;Buller, Benjamin;Jiang, Feng
侵袭性和血管生成是恶性胶质瘤的两个主要病理生理特征。抗血管生成治疗可增强肿瘤细胞的侵袭和转移。在本研究中，我们采用体内9L同种移植瘤动物模型和体外低氧细胞培养模型，利用RT2Profiler PCR芯片检测了胶质瘤细胞侵袭和血管生成相关基因的表达谱。还通过miRNA阵列获得了miRNA图谱。利用TargetScan数据库选择在mRNA阵列中有显著变化的基因来预测可能的调节mRNA表达的miRNAs，然后与miRNA阵列的结果进行匹配。基于这些标准，选择含有匹配miRNA miR-15b的NRP-2和具有匹配miRNA miR-152的基质金属蛋白酶-3进行进一步研究，以确定它们是否调节肿瘤微环境的变化，并影响胶质瘤的血管生成和侵袭。在9L胶质瘤细胞中存在NRP-2和MMP3的蛋白表达，并分别与miR-15b和miR-152的表达水平呈负相关。与9L胶质瘤细胞共培养的大鼠星形胶质细胞(原代和细胞系)与未共培养的星形胶质细胞相比，NRP-2、MMP3的表达显著增加，miR-15b、miR-152的表达显著降低。荧光素酶活性检测证实miR-15b和miR-152分别通过与NRP-2和MMP3转录本结合而抑制NRP-2和MMP3蛋白的表达。体外侵袭实验结果显示，miR-15b和miR-152可显著降低9L细胞的侵袭力。抗miR-15b和抗miR-152抑制剂可拮抗miR-15b和miR-152对肿瘤侵袭的抑制作用。体外小管形成实验结果显示，miR-15b可减少培养的内皮细胞的小管形成，而miR-152无此作用，而抗miR-15b抑制剂可拮抗miR-15b对小管形成的抑制作用。初步的通路研究表明，miR-15b和miR-152分别通过NRP-2和MMP-3失活9L细胞中的MEK-ERK通路。综上所述，我们目前的研究表明miR-15b通过NRP-2减少胶质瘤细胞的侵袭和血管生成，miR-152通过基质金属蛋白酶-3减少胶质瘤细胞的侵袭。
Invasion and angiogenesis are two major pathophysiological features of malignant gliomas. Anti-angiogenic treatment lead to enhanced tumor cell invasion and metastasis. In the current study, we tested invasion and angiogenesis related mRNA expression profiles of glioma cells via RT2Profiler PCR Array by employing an in vivo 9L homograft glioma tumor animal model and an in vitro hypoxic cell culture model. The miRNA profile was also obtained via miRNA array. Genes with mRNA expression that changed significantly in the mRNA array were selected to predict possible miRNAs that regulate mRNA expression using the TargetScan database, and were then matched with miRNA array results. Based on these criteria, NRP-2 with the matching miRNA miR-15b, and MMP-3 with the matching miRNA miR-152 were selected for further study, and to determine whether they regulate tumor microenviroment changes and affect glioma angiogenesis and invasion. The protein expression of NRP-2 and MMP-3 were verified in 9L glioma cells and were negatively correlated to miR-15b and miR-152 level, respectively. Rat astrocytes (primary and cell line), when co-cultured with 9L glioma cells, showed significantly elevated NRP-2, MMP-3 expression and reduced miR-15b, miR-152 expression compared to non co-cultured astrocytes. Luciferase activity assay confirmed that miR-15b and miR-152 attenuate expression of NRP-2 and MMP-3 protein by binding to NRP-2 and MMP-3 transcript, respectively. In vitro invasion assay data showed that miR-15b and miR-152 significantly decreased 9L cell invasiveness. Anti-miR-15b and anti-miR-152 inhibitors counteracted the inhibition of invasion caused by miR-15b and miR-152. In vitro tube formation assay data showed that miR-15b, but not miR-152, reduced tube formation in cultured endothelial cells, and anti-miR-15b inhibitor counteracted the inhibition of tube formation caused by miR-15b. A preliminary pathway study indicated that miR-15b and miR-152 deactivated the MEK-ERK pathway via NRP-2 and MMP-3 in 9L cells, respectively. In conclusion, our current study indicates that miR-15b reduces invasion of glioma cells and angiogenesis via NRP-2, and miR-152 reduces invasion of glioma cells through MMP-3.
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DOI：
10.1146/annurev.pathol.4.110807.092222
发表时间：
2009
期刊：
Annual review of pathology
影响因子：
--
作者：
Lee YS;Dutta A
通讯作者：
Dutta A
影响因子：
3.4
作者：
Di Leva G;Briskin D;Croce CM
通讯作者：
Croce CM
影响因子：
50.3
作者：
Blouw, B;Song, HQ;Bergers, G
通讯作者：
Bergers, G
影响因子：
11.5
作者：
Lamszus, K;Brockmann, MA;Westphal, M
通讯作者：
Westphal, M
影响因子：
8
作者：
Sternlicht, MD;Bissell, MJ;Werb, Z
通讯作者：
Werb, Z