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miR-361-3p 通过靶向 ARID3A 促进骨肉瘤细胞的肿瘤发生。

MiR-361-3p promotes tumorigenesis of osteosarcoma cells via targeting ARID3A.

机构信息

Department of Orthopaedics, The Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, Fujian 361101, PR China.

Basic Medical College of Heilongjiang University of Traditional Chinese Medicine, Harbin, Heilongjiang 150040, PR China.

出版信息

Tissue Cell. 2022 Jun;76:101759. doi: 10.1016/j.tice.2022.101759. Epub 2022 Feb 12.

Abstract

MiR-361-3p has been reported in several types of human cancer. However, the expression profile and biological functions of miR-361-3p in osteosarcoma remain uncovered. The expression profiles of miR-361-3p in osteosarcoma tissues and cell lines were evaluated using RT-qPCR. In 88 osteosarcoma patients, survival analysis was performed using Kaplan-Meier curves; while prognostic significance of miR-361-3p was analyzed using Cox regression analysis. The effects of miR-361-3p on cell proliferation, migration and invasion capacities were analyzed using CCK-8 and transwell assays. The target genes of miR-361-3p were assessed using luciferase reporter assay, RT-qPCR, Western blot and rescue experiments. Xenograft assay was conducted to test tumor growth ability. MiR-361-3p was found to be upregulated in human osteosarcoma tissues and cell lines. The expression of miR-361-3p was observed to be closely associated with TNM stage and lung metastasis. High expression of miR-361-3p was found to be capable of predicting poor clinical prognosis in osteosarcoma patients. Whilst overexpression of miR-361-3p was demonstrated to promote the proliferation, migration and invasion of osteosarcoma cells; knockdown of miR-361-3p was shown to exhibit an opposite inhibitory effect. Bioinformatics analysis and luciferase reporter assays confirmed that ARID3A is a direct target of miR-361-3p. Functional assays demonstrated that osteosarcoma cell proliferation, migration and invasion were promoted by miR-361-3p via negative regulation of ARID3A. Finally, overexpression of ARID3A was shown to partially reverse the tumor-promoting effect of miR-361-3p. Besides, in vivo assays revealed that miR-361-3p overexpression facilitated tumor growth in nude mice. In conclusion, this study indicates that miR-361-3p is a crucial prognostic biomarker of osteosarcoma, and that targeting of miR-361-3p/ARID3A axis may be a promising strategy in osteosarcoma therapy.

摘要

miR-361-3p 已在多种人类癌症中被报道。然而,miR-361-3p 在骨肉瘤中的表达谱和生物学功能仍未被揭示。本研究采用 RT-qPCR 评估了 miR-361-3p 在骨肉瘤组织和细胞系中的表达谱。对 88 例骨肉瘤患者进行生存分析,采用 Kaplan-Meier 曲线;采用 Cox 回归分析分析 miR-361-3p 的预后意义。采用 CCK-8 和 Transwell 实验分析 miR-361-3p 对细胞增殖、迁移和侵袭能力的影响。采用荧光素酶报告实验、RT-qPCR、Western blot 和挽救实验评估 miR-361-3p 的靶基因。进行异种移植实验以检测肿瘤生长能力。研究发现 miR-361-3p 在人骨肉瘤组织和细胞系中上调。miR-361-3p 的表达与 TNM 分期和肺转移密切相关。miR-361-3p 高表达可预测骨肉瘤患者的不良临床预后。过表达 miR-361-3p 促进骨肉瘤细胞的增殖、迁移和侵袭;而 miR-361-3p 敲低则表现出相反的抑制作用。生物信息学分析和荧光素酶报告实验证实 ARID3A 是 miR-361-3p 的直接靶基因。功能实验表明,miR-361-3p 通过负调控 ARID3A 促进骨肉瘤细胞的增殖、迁移和侵袭。最后,过表达 ARID3A 部分逆转了 miR-361-3p 的促肿瘤作用。此外,体内实验表明 miR-361-3p 过表达促进裸鼠肿瘤生长。综上所述,本研究表明 miR-361-3p 是骨肉瘤的重要预后生物标志物,靶向 miR-361-3p/ARID3A 轴可能是骨肉瘤治疗的一种有前途的策略。

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