■食管鳞状细胞癌(ESCC)是消化道恶性肿瘤,发病机制复杂。迫切需要寻找ESCC靶向治疗位点并探索其发病机制。胸腺素α原(PTMA)在多种肿瘤中异常表达,对肿瘤恶性进展具有显著的调节作用。然而,PTMA在ESCC中的调控作用和机制尚未见报道。
■我们首先检测了ESCC患者的PTMA表达,ESCC的皮下肿瘤异种移植模型,和ESCC细胞。随后,细胞转染抑制了ESCC细胞中PTMA的表达,细胞计数试剂盒-8(CCK-8)检测细胞增殖和凋亡,5-乙炔基-2'-脱氧尿苷(EdU)染色,流式细胞术,和Westernblot。二氯-二氢-荧光素二乙酸酯(DCFH-DA)测定法用于检测细胞中的活性氧(ROS)水平,和MitoSOX荧光探针,5,5\',6,6'-四氯-1,1',3,3'-四乙基-苯并咪唑基碳花青碘化物(JC-1)染色,线粒体复合物试剂盒,Westernblot检测线粒体氧化磷酸化的表达。接下来,使用免疫共沉淀(co-IP)和免疫荧光(IF)技术检测PTMA和高迁移率族蛋白1(HMGB1)之间的组合.最后,通过细胞转染抑制细胞中PTMA的表达和HMGB1的过表达,并通过相关实验确定PTMA和HMGB1结合对ESCC线粒体氧化磷酸化的调节作用。
■PTMA在ESCC中的表达异常升高。抑制ESCC细胞中PTMA的表达显著降低ESCC细胞的活性并增加其凋亡。此外,干扰PTMA可通过抑制线粒体氧化磷酸化诱导ESCC细胞ROS聚集,这可以通过与HMGB1结合来实现。
■PTMA与HMGB1结合以调节线粒体氧化磷酸化,从而影响ESCC的恶性进展。
UNASSIGNED: Esophageal squamous cell carcinoma (ESCC) is a malignant tumor of the digestive tract with complex pathogenesis. There is a pressing need to search for ESCC targeted therapy sites and explore its pathogenesis. Prothymosin alpha (PTMA) is abnormally expressed in numerous tumors and has a significant regulatory effect on tumor malignant progression. However, the regulatory role and mechanism of PTMA in ESCC have not yet been reported.
UNASSIGNED: We first detected the PTMA expression in ESCC patients, subcutaneous tumor xenograft models of ESCC, and ESCC cells. Subsequently, PTMA expression in ESCC cells was inhibited by cell transfection, and cell proliferation and apoptosis were detected by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2\'-deoxyuridine (EdU) staining, flow cytometry, and Western blot. A dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay was used to detect reactive oxygen species (ROS) level in cells, and MitoSOX fluorescent probe, 5,5\',6,6\'-tetrachloro-1,1\',3,3\'-tetraethyl-benzimidazolyl carbocyanine iodide (JC-1) staining, mitochondrial complex kit, and Western blot were used to detect the expression of mitochondrial oxidative phosphorylation. Next, the combination between PTMA and high mobility group box 1 (HMGB1) was detected using Co-immunoprecipitation (co-IP) and immunofluorescence (IF) techniques. Finally, the expression of PTMA was inhibited and the expression of HMGB1 was overexpressed in cells via cell transfection, and the regulatory effect of PTMA and HMGB1 binding on mitochondrial oxidative phosphorylation in ESCC was determined through related experiments.
UNASSIGNED: The expression of PTMA in ESCC was abnormally elevated. The inhibition of PTMA expression in ESCC cells significantly decreased the activity of ESCC cells and increased their apoptosis. Moreover, interference with PTMA can induce ROS aggregation in ESCC cells by inhibiting mitochondrial oxidative phosphorylation, which may be achieved by binding to HMGB1.
UNASSIGNED: PTMA binds to HMGB1 to regulate mitochondrial oxidative phosphorylation, thereby affecting the malignant progression of ESCC.