MiR-1224-5p Activates Autophagy, Cell Invasion and Inhibits Epithelial-to-Mesenchymal Transition in Osteosarcoma Cells by Directly Targeting PLK1 Through PI3K/AKT/mTOR Signaling Pathway.
MiR-1224-5p Activates Autophagy, Cell Invasion and Inhibits Epithelial-to-Mesenchymal Transition in Osteosarcoma Cells by Directly Targeting PLK1 Through PI3K/AKT/mTOR Signaling Pathway.
复制标题
miR-1224-5p通过直接通过PI3K/AKT/MTOR信号通路靶向PLK1，激活了骨肉瘤细胞中的自噬，细胞侵袭并抑制骨肉瘤细胞中的上皮过渡。
DOI：
10.2147/ott.s274451
复制
发表时间：
2020
影响因子：
4
通讯作者：
Chen K
中科院分区：
文献类型：
--
作者：
Jin B;Jin D;Zhuo Z;Zhang B;Chen K
骨肉瘤（osteosarcoma，OS）是最常见的恶性骨肿瘤之一，预后差。MiR-1224- 5 p在肿瘤中起重要作用，但其在OS中的作用及机制尚不清楚。采用qRT-PCR检测miR-1224- 5 p和PLK 1在OS细胞、癌旁组织和细胞系中的表达。采用双荧光素酶报告基因分析验证miR-1224- 5 p与PLK 1之间的相互作用。分别转染miR-1224 - 5 p模拟物、NC模拟物、pc-NC和PLK 1干预miR-1224 - 5 p和PLK 1的表达。采用MTT法、集落形成实验、Transwell法和流式细胞术观察细胞增殖、侵袭和凋亡情况。Western blot检测PLK 1、PI 3 K/AKT/mTOR信号通路相关蛋白、自噬相关蛋白、上皮-间质转化（EMT）相关蛋白的表达水平。我们发现在OS组织和细胞中miR-1224- 5 p表达下调，PLK 1表达上调。另一方面，进一步证实PLK 1是miR-1224- 5 p的靶基因。miR-1224- 5 p过表达可抑制OS细胞的增殖、侵袭，促进凋亡; PLK 1过表达可促进OS细胞的增殖、侵袭，抑制凋亡。在miR-1224- 5 p组（miR-1224- 5 p过表达）中，PI 3 K、AKT和mTOR蛋白磷酸化水平显著降低，而自噬活性显著激活，EMT程度显著降低。但是PLK 1组（PLK 1过表达）的结果相反。此外，miR-1224- 5 p的过表达逆转了PLK 1上调对OS细胞的影响。miR-1224- 5 p靶向PLK 1，抑制PI 3 K/AKT/mTOR信号通路，从而介导OS细胞的增殖、侵袭、凋亡、自噬和EMT。
Osteosarcoma (OS) is one of the most common malignant bone tumors with a poor overall prognosis. MiR-1224-5p plays an important role in cancer, but its function and mechanism in OS have not been studied. The expression of miR-1224-5p and PLK1 was detected by qRT-PCR in OS cells, adjacent tissues, and cell lines. Dual-luciferase reporter gene assay was used to verify the interaction between miR-1224-5p and PLK1. The expression of miR-1224-5p and PLK1 was intervened by transfection with miR-1224-5p mimic, NC mimic, pc-NC and PLK1, respectively. MTT, colony formation assay, Transwell and flow cytometry were used to observe the cell proliferation, invasion and apoptosis. Western blot was used to detect the expression levels of PLK1, PI3K/AKT/mTOR signaling pathway-related proteins, autophagy-related proteins, and epithelial-mesenchymal transition (EMT)-related proteins in the cells. We found that miR-1224-5p was down-regulated and PLK1 expression was up-regulated in OS tissues and cells. On the other hand, it is further confirmed that PLK1 was a target gene of miR-1224-5p. Overexpression of miR-1224-5p inhibited the proliferation, invasion while promoted the apoptosis of OS cells, whereas overexpression of PLK1 promoted the proliferation, invasion and inhibited the apoptosis of OS cells. In the miR-1224-5p group (overexpression of miR-1224-5p), PI3K, AKT, and mTOR protein phosphorylation levels were significantly reduced, while autophagic activity was significantly activated, and the degree of EMT was significantly reduced. But the results in the PLK1 group (overexpression of PLK1) were the opposite. In addition, overexpression of miR-1224-5p reversed the effect of PLK1 upregulation on OS cells. MiR-1224-5p targets PLK1 to inhibit PI3K/AKT/mTOR signaling pathway, thus mediating the proliferation, invasion, apoptosis, autophagy and EMT in OS cells.