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敲低 miR-384 通过靶向 SLBP 促进骨肉瘤 MG63 细胞的生长和转移。

Knocking down miR-384 promotes growth and metastasis of osteosarcoma MG63 cells by targeting SLBP.

机构信息

a Department of Orthopedics , China-Japan Union Hospital of Jilin University , Changchun , China.

b Department of Radiology , China-Japan Union Hospital of Jilin University , Changchun , China.

出版信息

Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):1458-1465. doi: 10.1080/21691401.2019.1601099.

Abstract

Osteosarcoma is a common malignant bone tumour in adolescents and old people, with highly invasive and metastatic features and poor prognosis. This study aimed to explore the role of miR-384 in osteosarcoma MG63 cells by targeting SLBP. Cell viability, migration and invasion, apoptosis, as well as apoptosis-related factors were evaluated by CCK-8 assay, Transwell assay, flow cytometer and Western blotting, respectively. Dual-luciferase reporter assay was used to determine the target of miR-384. SLBP level was analyzed using qRT-PCR and Western blotting. Important factors of MEK/ERK and PI3K/AKT signalling pathways were analyzed using Western blotting. We found that miR-384 was down-regulated in osteosarcoma tissue samples and cell lines (MG63, U2OS and OS732). miR-384 overexpression inhibited viability, migration and invasion, but promoted apoptosis of MG63 cells; whereas, miR-384 silence exhibited the contrary effects on MG63 cells. SLBP was a target of miR-384. Knockdown of SLBP reversed the promoting effect of miR-384 silence on cells, indicating that miR-384 silence promoted growth and metastasis of MG63 cells by up-regulating SLBP. In conclusion, knocking down miR-384 promoted the growth and metastasis of osteosarcoma MG63 cells by up-regulating SLBP. To conclude, miR-384-SLBP may be a potential therapeutic target for osteosarcoma therapy.

摘要

骨肉瘤是青少年和老年人常见的恶性骨肿瘤,具有侵袭性和转移性强、预后差的特点。本研究旨在通过靶向 SLBP 探讨 miR-384 在骨肉瘤 MG63 细胞中的作用。通过 CCK-8 测定、Transwell 测定、流式细胞术和 Western blot 分别评估细胞活力、迁移和侵袭、凋亡以及凋亡相关因子。双荧光素酶报告基因实验用于确定 miR-384 的靶标。使用 qRT-PCR 和 Western blot 分析 SLBP 水平。使用 Western blot 分析 MEK/ERK 和 PI3K/AKT 信号通路的重要因素。我们发现 miR-384 在骨肉瘤组织样本和细胞系(MG63、U2OS 和 OS732)中表达下调。miR-384 过表达抑制 MG63 细胞活力、迁移和侵袭,但促进细胞凋亡;而 miR-384 沉默对 MG63 细胞则表现出相反的作用。SLBP 是 miR-384 的靶标。SLBP 敲低逆转了 miR-384 沉默对细胞的促进作用,表明 miR-384 沉默通过上调 SLBP 促进 MG63 细胞的生长和转移。总之,敲低 miR-384 通过上调 SLBP 促进骨肉瘤 MG63 细胞的生长和转移。综上所述,miR-384-SLBP 可能是骨肉瘤治疗的潜在治疗靶点。

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