The role of microRNA-93 regulating angiopoietin2 in the formation of malignant pleural effusion.
The role of microRNA-93 regulating angiopoietin2 in the formation of malignant pleural effusion.
复制标题
microRNA-93调控血管生成素2在恶性胸腔积液形成中的作用
DOI：
10.1002/cam4.1000
复制
发表时间：
2017-05
期刊：
影响因子：
4
通讯作者：
Huang M
中科院分区：
文献类型：
--
作者：
Qian Q;Sun W;Zhu W;Liu Y;Ge A;Ma Y;Zhang Y;Zeng X;Huang M
MiRNAs在恶性胸腔积液(MPE)发生发展中的生物学作用尚不清楚。在本研究中，对两组不同预后的肺腺癌患者进行了miRNA微阵列分析。鉴定了miRNAs在MPE中的表达谱。利用定量聚合酶链式反应证实了miRNAs在体外的表达差异，并进一步分析了它们的生物学功能和相关的靶基因。通过在线数据库对miR-93的目的基因进行了预测，并对目的蛋白进行了检测。通过在线数据库对目的基因和特异性miRNA的结合位点进行分析。用荧光素酶报告基因检测结合位点的结合能力，用免疫印迹法检测目的基因蛋白。我们检测到107个表达差异的miRNAs(n=1110)，并证实两组患者(n=1084)miR-93和miR-146a的表达存在显著差异。通过调控人淋巴管内皮细胞(HLEC)和人脐静脉内皮细胞(HUVEC)miR-93的表达，我们发现miR-93的高表达抑制了细胞的迁移、增殖和血管生成。此外，miR-93不仅增加了细胞的凋亡率，而且还增加了G1期细胞的阻滞。通过荧光素酶报告基因分析和Western印迹分析，证实血管生成素2(Ang2)是miR-93的靶标。结果表明，miR-93对胸腔积液有抑制作用。MiR-93通过靶向Ang2调控血管生成和淋巴管生成，在MPE的发病机制中发挥作用。MIR-93/Ang2可能为癌症治疗提供潜在的新靶点。
The biological roles of miRNAs in the development of malignant pleural effusion (MPE) are unclear. In this study, the miRNA microarray analysis was performed in two different prognosis groups of lung adenocarcinoma patients. Expression profiles of miRNAs in MPEs were identified. With the help of quantification PCR, we confirmed the expression differences of miRNAs and further analyzed their biological functions and relative target genes in vitro. The target gene of miR‐93 was estimated by online database, and also, the protein was tested. The target gene and the binding sites of specific miRNA were estimated by online database. The combining capacity of binding sites was verified by luciferase reporter gene assay, and the target gene protein was tested by western blot. We detected 107 miRNAs with expression differences (n = 10) and confirmed significant expression differences in miR‐93 and miR‐146a in two groups of patients (n = 84). By manipulating miR‐93 expression of human lymphatic endothelial cells (HLEC) and human umbilical vein endothelial cells (HUVEC), we discovered that high expression of miR‐93 inhibited migration, proliferation, and angiogenesis. And also, miR‐93 increased not only apoptosis, but also G1 phase cell block. By using luciferase reporter gene assay and western blot, we confirmed that angiopoietin2 (Ang2) was the target of miR‐93. The data showed that miR‐93 has an inhibiting effect on pleural effusion. By targeting Ang2, miR‐93 regulates angiogenesis and lymphangiogenesis and plays a role in pathogenetic mechanism of MPE. MiR‐93/Ang2 may shed light on potential new targets in cancer treatment.