MicroRNA-17-5p 通过调节 PTEN 的表达来调控人骨肉瘤细胞的生长、迁移和侵袭。

MicroRNA-17-5p regulates the growth, migration and invasion of the human osteosarcoma cells by modulating the expression of PTEN.

机构信息

Department of Orthopedics, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200336.

出版信息

J BUON. 2020 Mar-Apr;25(2):1028-1034.

PMID:32521902

Abstract

PURPOSE

Osteosarcoma causes extensive human mortality and there is urgent need to develop novel therapies or to identify efficient therapeutic targets for its management. Herein the role and therapeutic potential of miR-17 was explored in osteosarcoma.

METHODS

The normal hFOB.19 cell line and the osteosarcoma cell lines SAOS-2, HOS, 143B, T1-73 and mG63 were used in the present study. The expression analysis of miR-17 was carried out by quantitative Real-Time polymerase chain reaction (qRT-PCR). Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA) was used for transfection. WST-1 assay was used for determination of cell proliferation and autophagy was detected by transmission electron microscopy (TEM). Wound healing and transwell assays were used for the determination of cell migration and invasion. Protein expression was determined by western blot analysis.

RESULTS

The expression of miR-17 was significantly elevated in all the osteosarcoma cells. Suppression of miR-17 resulted in decrease of the viability and colony formation of the SAOS-2 osteosarcoma cells. The inhibition of SAOS-2 cell proliferation upon miR-17 suppression was found to be due to induction of autophagy which was accompanied with enhancement in the expression of LC3B II and Beclin-1. Suppression of miR-17 was also accompanied by inhibition of the SAOS-2 cell migration and invasion. The in silico analysis showed that miR-187 targets PTEN in the SAOS-2 cells. The expression of PTEN was found to be downregulated in all the osteosarcoma cells and suppression of miR-17 expression caused enhancement in the expression of PTEN. Overexpression of miR-17 caused inhibition of the proliferation and colony formation of the SAOS-2 cells. Additionally, silencing of miR-17 could abolish the effects of miR-17 inhibition in the SAOS-2 cells.

CONCLUSION

MiR-17 may be proven a therapeutic target in the management of osteosarcoma.

摘要

目的

骨肉瘤导致了大量的人类死亡，因此迫切需要开发新的治疗方法或确定有效的治疗靶点来进行治疗。本研究旨在探讨 miR-17 在骨肉瘤中的作用和治疗潜力。

方法

本研究使用正常 hFOB.19 细胞系和骨肉瘤细胞系 SAOS-2、HOS、143B、T1-73 和 mG63。通过定量实时聚合酶链反应（qRT-PCR）进行 miR-17 的表达分析。使用 Lipofectamine 2000 试剂（Invitrogen，Carlsbad，CA，USA）进行转染。使用 WST-1 测定法测定细胞增殖，通过透射电子显微镜（TEM）检测自噬。使用划痕愈合和 Transwell 测定法测定细胞迁移和侵袭。通过 Western blot 分析测定蛋白表达。

结果

miR-17 在所有骨肉瘤细胞中的表达均显著升高。抑制 miR-17 导致 SAOS-2 骨肉瘤细胞活力和集落形成减少。抑制 miR-17 诱导自噬，从而导致 SAOS-2 细胞增殖抑制，这伴随着 LC3B II 和 Beclin-1 的表达增强。抑制 miR-17 还伴随着 SAOS-2 细胞迁移和侵袭的抑制。计算机分析表明，miR-187 是 SAOS-2 细胞中 PTEN 的靶标。所有骨肉瘤细胞中 PTEN 的表达均下调，抑制 miR-17 表达导致 PTEN 表达增强。过表达 miR-17 抑制了 SAOS-2 细胞的增殖和集落形成。此外，沉默 miR-17 可以消除 miR-17 抑制在 SAOS-2 细胞中的作用。