miR-224-5p inhibits proliferation, migration, and invasion by targeting PIK3R3/AKT3 in uveal melanoma
miR-224-5p inhibits proliferation, migration, and invasion by targeting PIK3R3/AKT3 in uveal melanoma
复制标题
DOI：
10.1002/jcb.28507
复制
发表时间：
2019-08-01
影响因子：
4
通讯作者：
Wang, Wenqi
中科院分区：
文献类型：
--
作者：
Li, Jianchang;Liu, Xiuming;Wang, Wenqi
葡萄膜黑色素瘤（UM）是成人最常见的原发性眼内恶性肿瘤。越来越多的研究发现UM中存在miRNAs（microRNAs）的异常表达，如miR-181、miR-20 a、miR-144、miR-146 a等。本研究旨在探讨miR-224- 5 p在UM中的生物学功能。采用Western blot分析和定量实时聚合酶链反应（qRT-PCR）分析30例肿瘤组织和配对的癌旁组织中miR-224- 5 p、PIK 3R 3和AKT 3的表达。采用细胞增殖实验、transwell实验和创伤愈合实验检测miR-224- 5 p对UM运动的影响。Western印迹分析和荧光素酶测定用于检测作为miR-224- 5 p下游靶标的PIK 3R 3和AKT 3的表达。Western blot和qRT-PCR检测结果显示，UM组织中miR-224- 5 p的表达较正常组织低，而PIK 3R 3和AKT 3的表达同时升高。上调miR-224- 5 p可显著抑制OCM-1A细胞的增殖、侵袭和迁移能力，并降低PIK 3R 3和AKT 3的表达。荧光素酶分析表明PIK 3R 3和AKT 3是miR-224- 5 p的下游靶点。此外，上调PIK 3R 3和AKT 3抑制了miR-224- 5 p诱导的OCM-1A细胞运动性抑制。因此，我们的研究证明miR-224- 5 p通过调节PIK 3R 3和AKT 3的表达参与UM细胞的增殖、侵袭和迁移。本研究还建立了miR-224- 5 p/PIK 3R 3/PI 3 K/AKT轴在UM进展中的调控机制，为进一步探索miR-224- 5 p作为UM患者治疗和诊断靶点提供了实验基础。
Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. Accumulating investigations have identified the aberrant expression of miRNAs (microRNAs) in UM, such as miR-181, miR-20a, miR-144, miR-146a. The purpose of this study is to investigate the biological function of miR-224-5p in UM. The expression of miR-224-5p, PIK3R3, and AKT3 in 30 tumor tissues and paired adjacent noncancerous tissues were analyzed using Western blot analysis and quantitative real-time polymerase chain reaction (qRT-PCR) assays. Cell proliferation assay, transwell assay, and wound healing assay were used to measure the effects of miR-224-5p on the motility of UM in vitro. Western blot analysis and luciferase assays were used to detect the expression of PIK3R3 and AKT3 as miR-224-5p downstream targets. The results of Western blot analysis and qRT-PCR assays indicated that the expression of miR-224-5p was lower in UM tissues compared to normal tissue, while the expression of PIK3R3 and AKT3 were simultaneously increased. Upregulation of miR-224-5p significantly inhibited capacities of proliferation, invasion, and migration of OCM-1A cells and decreased expression of PIK3R3 and AKT3. Luciferase assay demonstrated PIK3R3 and AKT3 as downstream targets of miR-224-5p. Moreover, upregulating PIK3R3 and AKT3 restrained miR-224-5p-induced inhibition of the motility of OCM-1A cells. Thus, our study proved that miR-224-5p was involved in proliferation, invasion, and migration of UM cells via regulation the expression of PIK3R3 and AKT3. And the results also established a miR-224-5p/PIK3R3/PI3K/AKT axis in the regulation of UM progression, providing an experimental basis for further exploring the miR-224-5p as a therapeutic and diagnosis target for patients with UM.