Overexpression of KH-type splicing regulatory protein regulates proliferation, migration, and implantation ability of osteosarcoma.
Overexpression of KH-type splicing regulatory protein regulates proliferation, migration, and implantation ability of osteosarcoma.
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DOI：
10.3892/ijo.2016.3601
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发表时间：
2016-09
影响因子：
5.2
通讯作者：
Srisomsap C
中科院分区：
文献类型：
--
作者：
Pruksakorn D;Teeyakasem P;Klangjorhor J;Chaiyawat P;Settakorn J;Diskul-Na-Ayudthaya P;Chokchaichamnankit D;Pothacharoen P;Srisomsap C
骨肉瘤是儿童和青少年常见的恶性骨肿瘤。目前的5年生存率约为60%，似乎正在达到一个平台。为了提高骨肉瘤的治疗效果，需要更好地了解肿瘤的发生和潜在的分子机制，以寻找可能的新的治疗靶点。本研究旨在利用蛋白质组学方法鉴定与骨肉瘤发病相关的蛋白质。对骨肉瘤（n=7）和松质骨成骨细胞（n=7）的原代细胞培养物中提取的蛋白质进行了研究。利用双向电泳技术和LC-MS/MS分析技术，成功地确定了7个差异表达蛋白点。在本研究中观察到4个上调蛋白和3个下调蛋白，其中选择KH型剪接调节蛋白（KSRP）进行进一步的探索。蛋白质印迹法检测KSRP在骨肉瘤细胞中的表达显著高于成骨细胞。另外，免疫组化结果显示，KSRP在实验组的独立病例骨肉瘤组织中也有高表达。更重要的是，在鸡胚绒毛尿囊膜实验中，KSRP沉默骨肉瘤细胞系显著降低细胞增殖、迁移能力以及植入和生长能力。总之，这些发现表明，KSRP在骨肉瘤发病机制的调控中起着重要作用，并作为骨肉瘤的潜在治疗靶点。
Osteosarcoma is a common malignant bone tumor in children and adolescents. The current 5-year survival rate is ~60% and that seems to be reaching a plateau. In order to improve treatment outcomes of osteosarcoma, a better understanding of tumorigenesis and underlying molecular mechanisms is required for searching out possible new treatment targets. This study aimed to identify the potential proteins involving the pathogenesis of osteosarcoma using a proteomics approach. Proteins extracted from primary cell culture of osteosarcoma (n=7) and osteoblasts of cancellous bone (n=7) were studied. Using 2-DE based proteomics and LC-MS/MS analysis, we successfully determined seven differentially expressed protein spots. Four upregulated proteins and three downregulated proteins were observed in this study in which KH-type splicing regulatory protein (KSRP) was selected for further exploration. KSRP was significantly upregulated in osteosarcoma cells compared to osteoblasts using western blot assay. In addition, immunohistochemistry demonstrated that KSRP was also highly expressed in osteosarcoma tissue of independent cases from the experimental group. More importantly, KSRP silencing of osteosarcoma cell lines significantly decreased cell proliferation, migration ability, as well as implantation and growth ability in chick chorioallantoic membrane assay. Taken together, these findings demonstrate, that KSRP plays important roles in regulatory controls of osteosarcoma pathogenesis and serves as a potentially therapeutic target of osteosarcoma.