Wu Hao, Yu Zhongjie, Chen Qi, Li Dong, Chen Wei
Trauma Center, Changzhou No.2 People's Hospital, the Affiliated Hospital of Nanjing Medical University, Changzhou, China.
J BUON. 2020 Mar-Apr;25(2):1013-1021.
The purpose of this study was to investigate the expression of microRNA (miRNA)-301a in osteosarcoma (OS) and its relationship with clinicopathological parameters and prognosis of patients with OS, and to further explore how it accelerates the progression of OS via modulating downstream target genes.
Quantitative real-time polymerase chain reaction (qRT-PCR) was used to examine the expression of miRNA-301a in 39 OS tumor tissue samples and adjacent ones, and the interplay between miRNA-301a and clinical indicators. The prognosis of patients with OS was analyzed. In addition, miRNA-301a overexpression vector was constructed to analyze the effect of miRNA-301a on the function of OS cells by cell counting kit-8 (CCK-8), transwell and cell wound healing assays. Finally, the potential mechanism was also investigated using luciferase reporter gene assay and cell recovery experiment.
qRT-PCR results revealed that miRNA-301a level in OS tumor tissue specimen was remarkably lower than that in adjacent tissue. Compared with patients with high expression of miRNA-301a, patients with low expression had a higher incidence of distant metastasis and lower overall survival. Compared with the negative control group (miR-NC group), cell proliferation and metastasis ability were remarkably decreased in the miRNA-301a mimics group. In addition, DEC2 expression was found remarkably elevated in OS cell lines and negatively correlated with miRNA-301a level. At the same time, cell recovery experiment demonstrated that there existed a mutual regulation between miRNA-301a and DEC2, the two of which could together promote the malignant progression of OS.
MiRNA-301a level was remarkably reduced both in OS tissues and cell line samples, and was confirmed to be associated with distant metastasis and poor prognosis of patients with OS. In addition, miRNA-301a was found to be able to inhibit malignant progression of OS through regulating DEC2.
本研究旨在探讨微小RNA（miRNA）-301a在骨肉瘤（OS）中的表达及其与骨肉瘤患者临床病理参数和预后的关系，并进一步探索其如何通过调节下游靶基因促进骨肉瘤进展。
采用定量实时聚合酶链反应（qRT-PCR）检测39例骨肉瘤肿瘤组织样本及其癌旁组织中miRNA-301a的表达，并分析miRNA-301a与临床指标之间的相互作用。分析骨肉瘤患者的预后。此外，构建miRNA-301a过表达载体，通过细胞计数试剂盒-8（CCK-8）、Transwell和细胞划痕愈合试验分析miRNA-301a对骨肉瘤细胞功能的影响。最后，采用荧光素酶报告基因试验和细胞回复实验研究其潜在机制。
qRT-PCR结果显示，骨肉瘤肿瘤组织标本中miRNA-301a水平显著低于癌旁组织。与miRNA-301a高表达患者相比，低表达患者远处转移发生率更高，总生存期更低。与阴性对照组（miR-NC组）相比，miRNA-301a模拟物组细胞增殖和转移能力显著降低。此外，发现DEC2在骨肉瘤细胞系中表达显著升高，且与miRNA-301a水平呈负相关。同时，细胞回复实验表明miRNA-301a与DEC2之间存在相互调节作用，二者共同促进骨肉瘤的恶性进展。
骨肉瘤组织和细胞系样本中miRNA-301a水平均显著降低，且证实其与骨肉瘤患者远处转移及不良预后相关。此外，发现miRNA-301a可通过调节DEC2抑制骨肉瘤的恶性进展。
