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微小RNA-17通过调节SASH1的表达促进骨肉瘤细胞的增殖和迁移并抑制其凋亡。

MicroRNA-17 promotes osteosarcoma cells proliferation and migration and inhibits apoptosis by regulating SASH1 expression.

作者信息

Wu Dajiang, Zhang Hao, Ji Fang, Ding Wenbin

机构信息

Department of Traumatic Orthopedics, Changhai Hospital, Shanghai, 200433, China.

Department of Traumatic Orthopedics, Changhai Hospital, Shanghai, 200433, China.

出版信息

Pathol Res Pract. 2019 Jan;215(1):115-120. doi: 10.1016/j.prp.2018.10.012. Epub 2018 Oct 22.

Abstract

MicroRNAs (miRNAs) are abnormally expressed in numerous diseases, which are intimately associated with cell proliferation, migration and invasion. Recent study indicated that miR-17 may be involved in regulating osteosarcoma (OS) occurrence and development, but its function and mechanism have not been reported. In this study, quantitative real-time PCR (qRT-PCR) was used to measure the expression of miR-17, and Western blotting assay was performed to measure the expressions of SAM and SH3 domain containing 1 (SASH1), phosphoinoinositide-3 kinase (PI3K), protein kinase B (AKT), Caspase3, Bcl-2 gene family (Bcl-2, Bax) and matrix metalloprotein (MMP-2, MMP-9) in MG-63 cells. Luciferase reporter assay was conducted to confirm the target of SASH1 by miR-17. Cell proliferation, migration, invasion and apoptosis assay was performed to investigate the role of miR-17 in OS cells. We found that the expression of miR-17 was significantly up-regulated in OS cell lines. MiR-17 inhibitor inhibited the proliferation ability, and induced apoptosis of OS cells. Besides, miR-17 inhibitor prevented the migration and invasion of OS cells. Further, we identified that SASH1 was a target gene of miR-17. In addition, knockdown of miR-17 increased the protein expression of SASH1, and regulate related genes of cell proliferation, invasion and anti-apoptosis in the downstream of OS cells. These findings indicated that miR-17 was over-expressed and promoted cell proliferation, migration and inhibited cell apoptosis by targeting SASH1 in OS cells.

摘要

微小RNA(miRNA)在众多疾病中表达异常,这与细胞增殖、迁移和侵袭密切相关。最近的研究表明,miR-17可能参与调节骨肉瘤(OS)的发生和发展,但其功能和机制尚未见报道。在本研究中,采用定量实时PCR(qRT-PCR)检测miR-17的表达,并通过蛋白质印迹法检测MG-63细胞中含SAM和SH3结构域蛋白1(SASH1)、磷酸肌醇-3激酶(PI3K)、蛋白激酶B(AKT)、半胱天冬酶3、Bcl-2基因家族(Bcl-2、Bax)和基质金属蛋白酶(MMP-2、MMP-9)的表达。进行荧光素酶报告基因检测以证实miR-17对SASH1的靶向作用。进行细胞增殖、迁移、侵袭和凋亡检测以研究miR-17在OS细胞中的作用。我们发现,miR-17在OS细胞系中的表达显著上调。miR-17抑制剂抑制了OS细胞的增殖能力并诱导其凋亡。此外,miR-17抑制剂还阻止了OS细胞的迁移和侵袭。进一步研究发现,SASH1是miR-17的靶基因。此外,敲低miR-17可增加SASH1的蛋白表达,并调节OS细胞下游与细胞增殖、侵袭和抗凋亡相关的基因。这些研究结果表明,miR-17在OS细胞中过表达,并通过靶向SASH1促进细胞增殖和迁移,抑制细胞凋亡。

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