Chen Shaogeng, He Rongqi, Lin Xianzuan, Zhang Wanfei, Chen Heshan, Xu Rongyu, Kang Mingqiang
Department of Thoracic Surgery, Fujian Medical University Union Hospital, Fuzhou, China.
Department of Thoracic Surgery, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, China.
J Thorac Dis. 2023 Mar 31;15(3):1302-1318. doi: 10.21037/jtd-23-143. Epub 2023 Mar 29.
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 () is abnormally expressed in numerous tumors and has a significant regulatory effect on tumor malignant progression. However, the regulatory role and mechanism of in ESCC have not yet been reported.
We first detected the expression in ESCC patients, subcutaneous tumor xenograft models of ESCC, and ESCC cells. Subsequently, 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 and high mobility group box 1 () was detected using Co-immunoprecipitation (co-IP) and immunofluorescence (IF) techniques. Finally, the expression of was inhibited and the expression of was overexpressed in cells via cell transfection, and the regulatory effect of and binding on mitochondrial oxidative phosphorylation in ESCC was determined through related experiments.
The expression of in ESCC was abnormally elevated. The inhibition of expression in ESCC cells significantly decreased the activity of ESCC cells and increased their apoptosis. Moreover, interference with can induce ROS aggregation in ESCC cells by inhibiting mitochondrial oxidative phosphorylation, which may be achieved by binding to .
binds to to regulate mitochondrial oxidative phosphorylation, thereby affecting the malignant progression of ESCC.
食管鳞状细胞癌（ESCC）是一种发病机制复杂的消化道恶性肿瘤。迫切需要寻找ESCC的靶向治疗位点并探索其发病机制。前胸腺素α（Prothymosin alpha，ProTα）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，ProTα在ESCC中的调节作用及机制尚未见报道。
我们首先检测了ProTα在ESCC患者、ESCC皮下肿瘤异种移植模型及ESCC细胞中的表达。随后，通过细胞转染抑制ESCC细胞中ProTα的表达，并采用细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）染色、流式细胞术和蛋白质免疫印迹法检测细胞增殖和凋亡情况。采用二氯二氢荧光素二乙酸酯（DCFH-DA）检测细胞内活性氧（ROS）水平，并用MitoSOX荧光探针、5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑羰花青碘化物（JC-1）染色、线粒体复合物试剂盒和蛋白质免疫印迹法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ProTα与高迁移率族蛋白B1（HMGB1）之间的结合情况。最后，通过细胞转染抑制细胞中ProTα的表达并过表达HMGB1的表达，通过相关实验确定ProTα与HMGB1结合对ESCC线粒体氧化磷酸化的调节作用。
ProTα在ESCC中的表达异常升高。抑制ESCC细胞中ProTα的表达可显著降低ESCC细胞活性并增加其凋亡。此外，干扰ProTα可通过抑制线粒体氧化磷酸化诱导ESCC细胞内ROS聚集，这可能是通过与HMGB1结合实现的。
ProTα与HMGB1结合调节线粒体氧化磷酸化，从而影响ESCC的恶性进展。
