Yang Rui, Ning Jing, Wang Hainan, Ma Hui, Cui Lijuan
Department of Hematology, General Hospital of Ningxia Medical University, Yinchuan, China.
Cell Biochem Biophys. 2025 Feb 17. doi: 10.1007/s12013-025-01692-8.
Acute myeloid leukemia (AML) is a challenging hematologic malignancy with a poor prognosis. STYX, a catalytically inactive phosphatase, is overexpressed in various cancers and has been shown to promote cellular proliferation. However, its clinical relevance and impact on AML cell behavior remain poorly understood. This study investigates the role of STYX in AML and elucidates its underlying molecular mechanisms. Peripheral blood samples were collected from 50 patients with AML and 25 healthy controls, and the expression of STYX and FBXW7 was analyzed using RT-qPCR and Western blot. THP-1 cells (AML cell line) were transfected with lentivirus vectors to overexpress STYX, FBXW7, or CCNE1. The effects of these proteins on THP-1 cell proliferation and apoptosis were assessed by RT-qPCR, Western blot, CCK-8, EdU, and TUNEL assays. Interactions between STYX and FBXW7, as well as FBXW7 and CCNE1, were confirmed via STRING analysis and endogenous co-immunoprecipitation (CO-IP). Furthermore, the ubiquitination level of CCNE1 was examined through immunoprecipitation (IP) and Western blot. Upregulated STYX mRNA and protein levels, along with downregulated FBXW7 mRNA and protein levels, were observed in peripheral blood samples from MLL-AF9 fusion gene-positive AML cases, with a negative correlation between STYX and FBXW7. Overexpression of STYX in AML cells increased cell viability, promoted proliferation, and inhibited apoptosis, thus accelerating AML progression. STYX overexpression also facilitated the interaction with FBXW7, downregulated FBXW7 expression, and impaired the ubiquitin-mediated degradation of CCNE1. FBXW7 overexpression reversed STYX-induced proliferation and apoptosis effects in AML cells, while CCNE1 overexpression counteracted the suppressive effects of FBXW7 on AML progression. STYX promotes AML proliferation by disrupting the ubiquitin degradation pathway of CCNE1 through its interaction with FBXW7, thereby accelerating disease progression. These findings indicate that targeting STYX may offer a promising therapeutic approach for AML.
急性髓系白血病(AML)是一种具有挑战性的血液系统恶性肿瘤,预后较差。STYX是一种催化失活的磷酸酶,在多种癌症中过表达,并已被证明可促进细胞增殖。然而,其临床相关性以及对AML细胞行为的影响仍知之甚少。本研究调查了STYX在AML中的作用,并阐明其潜在的分子机制。收集了50例AML患者和25例健康对照者的外周血样本,使用RT-qPCR和蛋白质印迹法分析STYX和FBXW7的表达。用慢病毒载体转染THP-1细胞(AML细胞系)以过表达STYX、FBXW7或CCNE1。通过RT-qPCR、蛋白质印迹法、CCK-8、EdU和TUNEL检测评估这些蛋白对THP-1细胞增殖和凋亡的影响。通过STRING分析和内源性免疫共沉淀(CO-IP)证实了STYX与FBXW7以及FBXW7与CCNE1之间的相互作用。此外,通过免疫沉淀(IP)和蛋白质印迹法检测CCNE1的泛素化水平。在MLL-AF9融合基因阳性AML病例的外周血样本中观察到STYX mRNA和蛋白水平上调,同时FBXW7 mRNA和蛋白水平下调,且STYX与FBXW7呈负相关。AML细胞中STYX的过表达增加了细胞活力,促进了增殖,并抑制了凋亡,从而加速了AML进展。STYX的过表达还促进了与FBXW7的相互作用,下调了FBXW7的表达,并损害了泛素介导的CCNE1降解。FBXW7的过表达逆转了STYX诱导的AML细胞增殖和凋亡作用,而CCNE1的过表达抵消了FBXW7对AML进展的抑制作用。STYX通过与FBXW7相互作用破坏CCNE1的泛素降解途径,从而促进AML增殖,进而加速疾病进展。这些发现表明,靶向STYX可能为AML提供一种有前景的治疗方法。