a Department of Orthopedics , The Second Xiangya Hospital, Central South University , Changsha , Hunan China.
b Department of Oncology , Xiangya Hospital, Central South University , Changsha , Hunan China.
Cancer Biol Ther. 2019;20(8):1141-1148. doi: 10.1080/15384047.2019.1598766. Epub 2019 Apr 12.
It remains unclear that long noncoding RNAs' role in cancer initiation and progression, including osteosarcoma. Long noncoding RNA LINC00963 was found to be participated in carcinogenesis and progression of osteosarcoma. However, the molecular mechanisms of LINC00963 engaged in osteosarcoma (OS) still needs to be explored. LINC00963 and miR-204-3p RNA expression levels were quantified by PCR in OS tissues and cells. CCK 8 assay, wound healing assay and transwell migration and invasion assay were chosen to assess cell growth, viability, migration, and invasion. Luciferase reporter assays were performed to verify direct interaction between LINC00963 and miR-204-3p and miR-204-3p and Fibronectin-1. Western blot was conducted to evaluate Fibronectin-1 expression in OS cells. LINC00963 was verified to be highly expressed in OS samples and cells. Specifically, elevated expression of LINC00963 was correlated with poor prognosis in patients. Furthermore, LINC00963 overexpression was found to promote proliferation, migration, and invasion . The luciferase reporter assay showed that LINC00963 can suppress miR-204-3p by directly binding miR-204-3p. Rescue experiment results indicated that function of LINC00963 in osteosarcoma was miR-204-3p dependant. Besides, we initially explored Fibronectin-1 (FN1) as the target of LINC00963/miR-204-3p axis in osteosarcoma. Our findings implied that LINC00963/miR-204-3p/FN1 can play an important role in proliferation and progression in osteosarcoma. LINC00963 has the potential to be a therapeutic target for osteosarcoma treatment.
长链非编码 RNA 在癌症发生和进展中的作用尚不清楚,包括骨肉瘤。研究发现长链非编码 RNA LINC00963 参与骨肉瘤的发生和进展。然而,LINC00963 参与骨肉瘤(OS)的分子机制仍有待探索。通过 PCR 定量检测 OS 组织和细胞中的 LINC00963 和 miR-204-3p RNA 表达水平。选择 CCK8 测定法、划痕愈合测定法和 Transwell 迁移和侵袭测定法评估细胞生长、活力、迁移和侵袭。荧光素酶报告测定法用于验证 LINC00963 和 miR-204-3p 以及 miR-204-3p 和 Fibronectin-1 之间的直接相互作用。Western blot 用于评估 OS 细胞中 Fibronectin-1 的表达。LINC00963 在 OS 样本和细胞中被证实高表达。具体来说,LINC00963 的高表达与患者的不良预后相关。此外,发现 LINC00963 过表达可促进增殖、迁移和侵袭。荧光素酶报告测定法表明,LINC00963 可通过直接结合 miR-204-3p 来抑制 miR-204-3p。 rescue 实验结果表明,LINC00963 在骨肉瘤中的作用依赖于 miR-204-3p。此外,我们初步探索了 Fibronectin-1 (FN1) 作为 LINC00963/miR-204-3p 轴在骨肉瘤中的靶点。我们的研究结果表明,LINC00963/miR-204-3p/FN1 可能在骨肉瘤的增殖和进展中发挥重要作用。LINC00963 有可能成为骨肉瘤治疗的潜在靶点。
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