Department of Spine Surgery, People's Hospital of Longhua, Affiliated Hospital of Southern Medical University, Guangdong 518000, Shenzhen, China.
Department of Spine Surgery, People's Hospital of Longhua, Affiliated Hospital of Southern Medical University, Guangdong 518000, Shenzhen, China.
Life Sci. 2021 Jul 15;277:119501. doi: 10.1016/j.lfs.2021.119501. Epub 2021 Apr 15.
The present study evaluated the functions of Piperlongumine (PL) in osteosarcoma (OS) cell growth and metastasis both in vitro and in vivo.
MTT assay was conducted to test the cytotoxic effects of PL on the human osteoblasts line HFOB1.19 and the human normal chondrocyte line C28/I2T. FITC-Annexin V and propidium iodide (PI) were used to examine cell apoptosis. The migration, invasion and relative epithelial-mesenchymal transition were examined by Transwell assay and Western blotting. Reverse transcription-quantitative PCR (RT-qPCR) was performed to analyze the cytokine signaling 3 (SOCS3) mRNA expression. TargetScan database was used to predict the target of SOCS3. The binding association between miR-30d-5p and SOCS3 in U2OS and MG63 cells was evaluated by the dual-luciferase reporter assay. A xenograft model was constructed to evaluate the effect of PL on OS cell growth in vivo.
The results revealed that PL inhibited the growth, migration, invasion, epithelial-mesenchymal transition, and promoted the apoptosis of OS cells dose-dependently. In addition, PL upregulated the protein levels of suppressor of SOCS3, while it inactivated the JAK2/STAT3 pathway, which was accompanied by a decreased level of microRNA (miR)-30d-5p. Furthermore, SOCS3was confirmed as a novel target of miR-30d-5p. Overexpression of miR-30d-5p not only led to decreased expression of SOCS3, but also dampened the antitumor effect of PL on OS.
The present data demonstrated that PL inhibited the progression of OS via downregulation of the SOCS3-mediated JAK2/STAT3 pathway by inhibiting miR-30d-5p.
本研究评估了荜茇酰胺（PL）在体外和体内对骨肉瘤（OS）细胞生长和转移的作用。
MTT 法检测 PL 对人成骨细胞系 HFOB1.19 和人正常软骨细胞系 C28/I2T 的细胞毒性作用。FITC-Annexin V 和碘化丙啶（PI）检测细胞凋亡。Transwell 检测和 Western blot 检测细胞迁移、侵袭和相对上皮-间充质转化。逆转录定量 PCR（RT-qPCR）分析细胞因子信号转导 3（SOCS3）mRNA 表达。TargetScan 数据库预测 SOCS3 的靶点。双荧光素酶报告基因检测 U2OS 和 MG63 细胞中 miR-30d-5p 与 SOCS3 的结合关系。构建异种移植模型评估 PL 对 OS 细胞体内生长的影响。
结果表明，PL 呈剂量依赖性抑制 OS 细胞生长、迁移、侵袭、上皮-间充质转化，促进细胞凋亡。此外，PL 上调 SOCS3 蛋白水平，同时激活 JAK2/STAT3 通路，伴有 miR-30d-5p 水平降低。进一步证实 SOCS3 是 miR-30d-5p 的新靶点。miR-30d-5p 的过表达不仅导致 SOCS3 表达降低，还减弱了 PL 对 OS 的抗肿瘤作用。
本研究数据表明，PL 通过抑制 miR-30d-5p 抑制 SOCS3 介导的 JAK2/STAT3 通路，抑制骨肉瘤的进展。
