Departments of Radiology, China-Japan Union Hospital of Jilin University, Changchun, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(3):972-981. doi: 10.26355/eurrev_201902_16984.
MicroRNAs are endogenous, non-coding small RNAs that are capable of regulating biological and pathological processes. Previous studies have shown that microRNA-1269a serves as an oncogene. However, the role of microRNA-1269a in the pathogenesis of osteosarcoma (OS) has not been reported. The aim of this work was to investigate the expression characteristics of microRNA-1269a in OS and to further study its regulatory effects on the malignant progression of OS.
The expression of microRNA-1269a in 61 pairs of OS tissues and para-cancerous tissues was detected by quantitative Real Time-polymerase Chain Reaction (qRT-PCR). Chi-square test was used to analyze the relationship between microRNA-1269a expression and the characteristics of OS patients, including age, sex, clinical stage and distant metastasis. Subsequently, microRNA-1269a expression in OS cell lines was detected as well. After knockdown of microRNA-1269a by constructing relevant small interference RNA, biological performances of MG63 and U2OS cells were accessed by cell counting kit-8 (CCK-8), colony formation and transwell assay. Meanwhile, the protein expressions of key genes in the EMT/Smad pathway were detected by Western blot. Finally, si-TGF-β1 (transforming growth factor-β1) was transfected into OS cells, and cell migration and invasion were detected by transwell assay.
MicroRNA-1269a was highly expressed in OS tissues compared with para-cancerous tissues. High expression of microRNA-1269a was positively correlated with young OS patients and high rate of distant metastasis, whereas was not correlated with age, sex and Enneking stage. Kaplan-Meier survival curves showed that high expression of microRNA-1269a was significantly associated with poor prognosis of OS. The knockdown of microRNA-1269a in MG63 and U2OS cells significantly inhibited cell proliferation, migration and invasion. Meanwhile, microRNA-1269a knockdown in OS cells markedly downregulated the expressions of TGF-β1, p-Smad2, p-Smad3, N-cad, Vimentin and MMP9. Furthermore, TGF-β1 knockdown remarkably decreased migratory and invasive abilities of OS cells.
MicroRNA-1269a is highly expressed in OS, which is remarkably correlated with tumor stage, distant metastasis and poor prognosis of OS. In addition, microRNA-1269a promotes the malignant progression of OS by regulating TGF-β1 expression.
microRNAs 是一种内源性、非编码的小 RNA，能够调节生物和病理过程。先前的研究表明，microRNA-1269a 是一种致癌基因。然而，microRNA-1269a 在骨肉瘤（OS）发病机制中的作用尚未被报道。本研究旨在探讨 microRNA-1269a 在 OS 中的表达特征，并进一步研究其对 OS 恶性进展的调节作用。
采用实时定量聚合酶链反应（qRT-PCR）检测 61 对 OS 组织和癌旁组织中 microRNA-1269a 的表达。采用卡方检验分析 microRNA-1269a 表达与 OS 患者年龄、性别、临床分期和远处转移等特征的关系。随后，检测 OS 细胞系中 microRNA-1269a 的表达。通过构建相关小干扰 RNA 使 microRNA-1269a 沉默，用细胞计数试剂盒-8（CCK-8）、集落形成和 Transwell 检测 MG63 和 U2OS 细胞的生物学性能。同时，用 Western blot 检测 EMT/Smad 通路中关键基因的蛋白表达。最后，转染 OS 细胞 si-TGF-β1（转化生长因子-β1），用 Transwell 检测细胞迁移和侵袭。
与癌旁组织相比，microRNA-1269a 在 OS 组织中高表达。microRNA-1269a 的高表达与年轻的 OS 患者和高远处转移率呈正相关，而与年龄、性别和 Enneking 分期无关。Kaplan-Meier 生存曲线表明，microRNA-1269a 的高表达与 OS 的不良预后显著相关。在 MG63 和 U2OS 细胞中敲低 microRNA-1269a 显著抑制细胞增殖、迁移和侵袭。同时，OS 细胞中 microRNA-1269a 的敲低显著下调了 TGF-β1、p-Smad2、p-Smad3、N-cad、Vimentin 和 MMP9 的表达。此外，TGF-β1 的敲低显著降低了 OS 细胞的迁移和侵袭能力。
microRNA-1269a 在 OS 中高表达，与肿瘤分期、远处转移和 OS 的不良预后显著相关。此外，microRNA-1269a 通过调节 TGF-β1 的表达促进 OS 的恶性进展。
