Department of Emergency Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Department of Orthopedic Department, The Second Affiliated Hospital of Harbin Medical University, 246 Xuefu Road, Nangang District, Harbin, 150001, Heilongjiang, China.
J Orthop Surg Res. 2023 Feb 23;18(1):132. doi: 10.1186/s13018-023-03615-w.
Osteosarcoma is a common primary bone malignancy prevalent among adolescents and young adults. PTEN-induced kinase 1 (PINK1) regulates Parkinson's disease, but its role in cancers is unknown.
This study was designed to analyze the mechanism by which PINK1 affects osteosarcoma using bioinformatics and cell experiments.
The gene expression profiles were downloaded from the TARGET database. Several online databases were used to analyze the expression and protein‒protein interaction networks. CCK-8 cell viability assays and cisplatin treatment were used to assess cell activity with or without cisplatin treatment. Acridine orange/ethidium bromide (AO/EB) fluorescence staining was used to calculate the percentage of apoptotic cells.
Through bioinformatics analysis, we found that high expression of PINK1 was associated with poor prognosis in patients with osteosarcoma, and PINK1 inhibited apoptosis and promoted proliferation pathways. Next, we found that both PINK1 mRNA and protein levels were upregulated in osteosarcoma tissues. Additionally, we found that PTEN was reduced, while FOXO3a was markedly increased in osteosarcoma, suggesting that FOXO3a and not PTEN induced the overexpression of PINK1. CCK-8 and clonogenic assays showed that the knockdown of PINK1 decreased the growth of U2OS osteosarcoma cells. Ki67 immunofluorescence staining revealed that reduced cell proliferation in U2OS cells resulted in the depletion of PINK1. In addition, our AO/EB staining results indicated that the knockdown of PINK1 resulted in an increase in apoptotic cells and increased the levels of cleaved caspase-3. Furthermore, our experiments revealed that cisplatin promotes OS cell apoptosis by downregulating PINK1.
Collectively, our findings demonstrate that PINK1 is crucially involved in osteosarcoma and suggests that it can promote the apoptosis of OS cells as the downstream target gene of cisplatin.
骨肉瘤是一种常见的原发性骨恶性肿瘤，主要发生于青少年和年轻成年人。PTEN 诱导的激酶 1（PINK1）调节帕金森病，但它在癌症中的作用尚不清楚。
本研究旨在通过生物信息学和细胞实验分析 PINK1 影响骨肉瘤的机制。
从 TARGET 数据库中下载基因表达谱。使用多个在线数据库分析表达和蛋白质相互作用网络。用 CCK-8 细胞活力检测和顺铂处理来评估细胞活性，包括有无顺铂处理。用吖啶橙/溴化乙锭（AO/EB）荧光染色来计算凋亡细胞的百分比。
通过生物信息学分析，我们发现 PINK1 高表达与骨肉瘤患者的预后不良相关，并且 PINK1 抑制凋亡并促进增殖途径。接下来，我们发现骨肉瘤组织中 PINK1 的 mRNA 和蛋白水平均上调。此外，我们发现骨肉瘤中 PTEN 减少，而 FOXO3a 明显增加，表明 FOXO3a 而不是 PTEN 诱导 PINK1 的过表达。CCK-8 和集落形成实验表明，PINK1 的敲低降低了 U2OS 骨肉瘤细胞的生长。Ki67 免疫荧光染色显示，U2OS 细胞中细胞增殖减少导致 PINK1 耗竭。此外，我们的 AO/EB 染色结果表明，PINK1 的敲低导致凋亡细胞增加，并增加了 cleaved caspase-3 的水平。此外，我们的实验表明，顺铂通过下调 PINK1 促进 OS 细胞凋亡。
综上所述，我们的研究结果表明 PINK1 与骨肉瘤密切相关，并表明它可以作为顺铂的下游靶基因促进 OS 细胞的凋亡。
