Hang Jing, Ouyang Hanqiang, Wei Feng, Zhong Qihang, Yuan Wanqiong, Jiang Liang, Liu Zhongjun
Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, China.
Key Laboratory of Assisted Reproduction, Ministry of Education, Beijing, China.
Front Oncol. 2022 Sep 30;12:941046. doi: 10.3389/fonc.2022.941046. eCollection 2022.
Chordoma is a slow-growing but malignant subtype of bone sarcoma with relatively high recurrence rates and high resistance to chemotherapy. It is urgent to understand the underlying regulatory networks to determine more effective potential targets. Phosphorylative regulation is currently regarded as playing a significant role in tumorigenesis, and the use of tyrosine kinase inhibitors in clinical practice has yielded new promise for the treatment of a variety of sarcoma types.
We performed comprehensive proteomic and phosphoproteomic analyses of chordoma using four-dimensional label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS) and bioinformatics analysis. The potential aberrantly expressed kinases and their functions were validated using western blotting and CCK-8 assays.
Compared with paired normal muscle tissues, 1,139 differentially expressed proteins (DEPs) and 776 differentially phosphorylated proteins (DPPs) were identified in chordoma tumor tissues. The developmentally significant Wnt-signaling pathway and oxidative phosphorylation were aberrant in chordoma. Moreover, we predicted three kinases (AURA, CDK9, and MOK) with elevated activity by kinase-pathway network analysis (KiPNA) and verified their increased expression levels. The knockdown of these kinases markedly suppressed chordoma cell growth, and this was also the case for cells treated with the CDK9 inhibitor AZD4573. We additionally examined 208 proteins whose expression and phosphorylation levels were synergetically altered.
We herein depicted the collective protein profiles of chordomas, providing insight into chordomagenesis and the potential development of new therapeutic targets.
脊索瘤是一种生长缓慢但恶性的骨肉瘤亚型，复发率相对较高且对化疗耐药。了解其潜在的调控网络以确定更有效的潜在靶点迫在眉睫。磷酸化调节目前被认为在肿瘤发生中起重要作用，临床实践中使用酪氨酸激酶抑制剂为多种肉瘤类型的治疗带来了新希望。
我们使用四维无标记液相色谱 - 串联质谱（LC-MS/MS）和生物信息学分析对脊索瘤进行了全面的蛋白质组学和磷酸化蛋白质组学分析。使用蛋白质印迹法和CCK-8测定法验证了潜在异常表达的激酶及其功能。
与配对的正常肌肉组织相比，在脊索瘤肿瘤组织中鉴定出1139种差异表达蛋白（DEP）和776种差异磷酸化蛋白（DPP）。在脊索瘤中，具有重要发育意义的Wnt信号通路和氧化磷酸化异常。此外，通过激酶 - 通路网络分析（KiPNA）预测了三种活性升高的激酶（AURA、CDK9和MOK），并验证了它们表达水平的增加。敲低这些激酶显著抑制了脊索瘤细胞的生长，用CDK9抑制剂AZD4573处理的细胞也是如此。我们还检测了208种表达和磷酸化水平协同改变的蛋白质。
我们在此描绘了脊索瘤的整体蛋白质谱，为脊索瘤的发生机制及新治疗靶点的潜在开发提供了见解。
