Zhang Tao, Jing Li, Li Hong, Ding Linchao, Ai Dongdong, Lyu Jianxin, Zhong Lianjin
Key Laboratory of Laboratory Medicine, Ministry of Education, Zhejiang Provincial Key Laboratory of Medical Genetics, College of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, P.R. China.
Oncol Lett. 2017 Jul;14(1):111-118. doi: 10.3892/ol.2017.6102. Epub 2017 Apr 28.
The results of our previous study revealed that microRNA (miRNA/miR)-4530 was upregulated in the serum of patients with diabetic retinopathy. The TargetScan miRNA database was used to identify potential targets of miR-4530 and vasohibin-1 (VASH1) was predicted as one of the targets. The results of our previous study demonstrated that miR-4530 was able to promote angiogenesis in human umbilical vein endothelial cells. Therefore, suppressing miR-4530 may be a potentially novel approach towards inhibiting tumor angiogenesis. The present study aimed to investigate the function of miR-4530 and determine whether miR-4530 was able to regulate angiogenesis in breast carcinoma cells by targeting VASH1. MDA-MB-231 and MCF-7 cells were transfected with miR-4530 precursor, anti-miR-4530 and empty vector plasmids. The expression levels of miRNA and mRNA were detected using the reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The expression levels of protein were detected using western blotting. Dual-luciferase reporter assays were used to identify the target of miR-4530. Furthermore, cell proliferation, cell cycle, apoptosis and tube formation assays were used to investigate the function of miR-4530 . Nude mice were used in a subcutaneous tumor model study. The results of the present study demonstrated that miR-4530 significantly suppressed proliferation and promoted apoptosis of breast carcinoma cells. In addition, miR-4530 expression promoted angiogenesis . Results from the western blotting and RT-qPCR revealed that VASH1 was significantly downregulated by miR-4530 in breast carcinoma cells. The results of the present study suggest that miR-4530 promotes angiogenesis, inhibits proliferation and induces apoptosis in breast carcinoma cells by suppressing the expression of VASH1.
我们之前的研究结果显示，微小RNA（miRNA/miR）-4530在糖尿病视网膜病变患者的血清中表达上调。利用TargetScan miRNA数据库来识别miR-4530的潜在靶标，血管抑制素-1（VASH1）被预测为其中一个靶标。我们之前的研究结果表明，miR-4530能够促进人脐静脉内皮细胞的血管生成。因此，抑制miR-4530可能是一种潜在的抑制肿瘤血管生成的新方法。本研究旨在探讨miR-4530的功能，并确定miR-4530是否能够通过靶向VASH1来调节乳腺癌细胞中的血管生成。将miR-4530前体、抗miR-4530和空载体质粒转染至MDA-MB-231和MCF-7细胞。使用逆转录-定量聚合酶链反应（RT-qPCR）检测miRNA和mRNA的表达水平。使用蛋白质印迹法检测蛋白质的表达水平。采用双荧光素酶报告基因测定法来鉴定miR-4530的靶标。此外，利用细胞增殖、细胞周期、凋亡和管形成测定法来研究miR-4530的功能。在皮下肿瘤模型研究中使用裸鼠。本研究结果表明，miR-4530显著抑制乳腺癌细胞的增殖并促进其凋亡。此外，miR-4530的表达促进血管生成。蛋白质印迹法和RT-qPCR的结果显示，miR-4530在乳腺癌细胞中显著下调VASH1的表达。本研究结果表明，miR-4530通过抑制VASH1的表达来促进乳腺癌细胞中的血管生成、抑制增殖并诱导凋亡。
