| 黄崇新,吕波,王跃,等.PLK1抑制剂对骨肉瘤细胞增殖的影响及其机制研究.骨科,2016,7(5): 358-363. |
| PLK1抑制剂对骨肉瘤细胞增殖的影响及其机制研究 |
| Effect of PLK1 inhibitor on proliferation of osteosarcoma cells and its mechanism |
| 投稿时间：2016-02-19 |
| DOI：10.3969/j.issn.1674-8573.2016.05.014 |
| 中文关键词: 骨肉瘤 Polo样激酶1 蛋白激酶抑制剂 p53 生存分析 |
| 英文关键词: Osteosarcoma Polo-like kinase 1 Protein kinase inhibitors p53 Survival analysis |
| 基金项目:国家自然科学青年基金（81300964） |
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| 中文摘要: |
| 目的 探讨Polo样激酶1（polo-like kinase 1, PLK1）的小分子抑制剂NMS-P937对骨肉瘤细胞增殖的影响及其作用机制，并分析PLK1蛋白水平与临床骨肉瘤患者预后的关系。方法 本研究采用HOS、Saos-2骨肉瘤细胞系以及人成骨细胞系HOB-C作为实验细胞，通过Western Blot和荧光定量PCR检测PLK1在不同细胞系中蛋白和mRNA的表达水平；分别用不同浓度（0.01、0.05、0.1、0.2、0.5 μmol/L）的NMS-P937干预骨肉瘤细胞，采用MTT比色法检测细胞增殖情况，流式细胞术检测不同浓度NMS-P937对骨肉瘤细胞系细胞周期的调控作用；免疫共沉淀技术检测PLK1蛋白与p53蛋白的相互作用情况；通过对骨肉瘤患者的组织芯片分析及随访，分析PLK1蛋白表达水平与临床预后的关系。结果 研究结果表明与人成骨细胞系HOB-C细胞相比，HOS、Saos-2骨肉瘤细胞系中PLK1蛋白和mRNA表达水平更高；NMS-P937可有效抑制骨肉瘤细胞的增殖，且经过NMS-P937处理后，S、G2/M期的骨肉瘤细胞Saos-2显著升高，G0/G1期细胞显著降低；细胞免疫共沉淀结果表明PLK1与p53蛋白存在相互作用；生存分析显示PLK1的表达水平与患者生存率呈负相关。结论 PLK1抑制剂可抑制骨肉瘤细胞的增殖，这可能与PLK1通过调节p53蛋白参与细胞周期转换过程有关，此外，PLK1蛋白水平的高度表达与患者不良预后密切相关，提示PLK1蛋白可作为临床潜在治疗骨肉瘤患者的有效靶点。 |
| 英文摘要: |
| Objective To investigate the effect and mechanism of NMS-P937, a small molecule inhibitor of PLK1 on the proliferation of human osteosarcoma cells, and the relationship between the level of PLK1 and the prognosis of patients with osteosarcoma. Methods In this study, HOS, Saos-2 osteosarcoma cell lines and human osteoblastic cell line HOB-C were used as experimental samples. The protein and mRNA levels of PLK1 in various cell lines were detected by Western blotting and q-PCR respectively. Different concentrations (0.01, 0.05, 0.1, 0.2, 0.5 mol/L) of NMS-P937 were used to interfere osteosarcoma cells respectively. The inhibitory effects of NMS-P937 on proliferation were estimated by virtue of MTT assay, and regulation of cell cycle by NMS-P937 was verified applying flow cytometry. Association between PLK1 and p53 protein was tested by co-immunoprecipitation technology. Relationship between clinical prognosis and PLK1 protein expression level was analyzed through tissue array information by Kaplan-Meier. Results Our results indicated that PLK1 protein and mRNA expression levels were higher in HOS and Saos-2 cells than in HOB-c cells. The NMS-P937 inhibitor could effectively suppress the PLK1 expression level. Through treatment with NMS-P937 inhibitor, the number of Saos-2 cells in S and G2/M phases was elevated significantly and that in G0/G1 phase decreased evidently. Co-immunoprecipitation indicated association between PLK1 and p53 protein. Survival analysis showed a negative correlation between PLK1 protein expression level and patients’ survival rate. Conclusion The application of PLK1 inhibitor can inhibit the proliferation of osteosarcoma cells, which may be associated with the negative role of PLK1 in regulating p53 involved in cell cycle transformation. Besides, high PLK1 protein expression level affects patients’ prognosis, indicating PLK1 protein may exert potential target role in treating clinical osteosarcoma patients. |
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