Department of Pharmacy, Gansu Provincial Hospital, Lanzhou, Gansu, China.
Department III of Orthopedic, Gansu Provincial Hospital, Lanzhou, Gansu, China.
Yonsei Med J. 2019 Sep;60(9):832-841. doi: 10.3349/ymj.2019.60.9.832.
Epirubicin is one of the most effective drugs against osteosarcoma. miR-1301 is involved in the occurrence and development of osteosarcoma. Whether miR-1301 is responsible for the chemosensitivity of osteosarcoma cells to epirubicin remains largely unknown.
U2OS and SAOS-2 cells were treated with various concentrations of epirubicin. Flow cytometry was employed to evaluate cell apoptotic rate. Cell proliferation was measured by Cell Counting Kit-8 assay. Western blot and quantitative real-time polymerase chain reaction were utilized to detect the expressions of B-cell lymphoma-2 (Bcl-2), Bcl-2 assaciated X protein (Bax), cleaved-caspase-3, cleaved-poly (ADP-ribose) polymerases (PARP1), TP53-regulated inhibitor of apoptosis 1 (TRIAP1), and microRNA-1301 (miR-1301). The relationship between miR-1301 and TRIAP1 was determined by luciferase reporter assay.
Epirubicin inhibited proliferation in a dose-dependent manner, induced apoptosis, decreased the expression of Bcl-2, and increased the expressions of Bax, cleaved-caspase-3, and cleaved-PARP1 in osteosarcoma cells. miR-1301 was downregulated in U2OS and SAOS-2 cells. Importantly, epirubicin significantly increased the levels of miR-1301. Overexpression of miR-1301 suppressed proliferation and promoted apoptosis. Interestingly, those effects were enhanced by epirubicin. In contrast, miR-1301 depletion attenuated the epirubicin-mediated anti-osteosarcoma effect. miR-1301 negatively regulated the expression of TRIAP1 in U2OS and SAOS-2 cells. Furthermore, epirubicin inhibited the mRNA and protein levels of TRIAP1 by upregulating miR-1301 levels. Epirubicin suppressed cell proliferation by downregulating TRIAP1.
miR-1301 was implicated in the chemosensitivity of osteosarcoma to epirubicin by modulating TRIAP1.
表阿霉素是治疗骨肉瘤最有效的药物之一。miR-1301 参与骨肉瘤的发生和发展。miR-1301 是否负责骨肉瘤细胞对表阿霉素的化疗敏感性在很大程度上尚不清楚。
用不同浓度的表阿霉素处理 U2OS 和 SAOS-2 细胞。采用流式细胞术评估细胞凋亡率。通过细胞计数试剂盒-8 法测量细胞增殖。Western blot 和实时定量聚合酶链反应检测 B 细胞淋巴瘤-2（Bcl-2）、Bcl-2 相关 X 蛋白（Bax）、cleaved-caspase-3、cleaved-poly（ADP-ribose）聚合酶（PARP1）、TP53 调节的凋亡抑制剂 1（TRIAP1）和 microRNA-1301（miR-1301）的表达。通过荧光素酶报告基因测定确定 miR-1301 与 TRIAP1 之间的关系。
表阿霉素呈剂量依赖性抑制增殖，诱导凋亡，降低骨肉瘤细胞中 Bcl-2 的表达，增加 Bax、cleaved-caspase-3 和 cleaved-PARP1 的表达。U2OS 和 SAOS-2 细胞中 miR-1301 下调。重要的是，表阿霉素显著增加了 miR-1301 的水平。过表达 miR-1301 抑制增殖并促进凋亡。有趣的是，这些作用被表阿霉素增强。相反，miR-1301 耗竭减弱了表阿霉素介导的抗骨肉瘤作用。miR-1301 负调控 U2OS 和 SAOS-2 细胞中 TRIAP1 的表达。此外，通过上调 miR-1301 水平，表阿霉素抑制 TRIAP1 的 mRNA 和蛋白水平。表阿霉素通过下调 TRIAP1 抑制细胞增殖。
miR-1301 通过调节 TRIAP1 参与骨肉瘤对表阿霉素的化疗敏感性。
