Background: TSTA3 gene encodes an enzyme responsible for synthesis of GDP-L-fucose as the only donor in fucosylation. This study was designed to explore clinical value, function and underlying mechanism of TSTA3 in the development of esophageal squamous cell carcinoma (ESCC)./nMethods: Whole genomic sequencing data from 663 ESCC patients and RNA sequencing data from 155 ESCC patients were used to analyze the copy number variation and mRNA expression of TSTA3 respectively. Immunohistochemistry based or not based on the tissue microarrays was used to detect its protein expression. Transwell assay and in vivo metastasis assay were used to study the effect of TSTA3 on invasion and metastasis of ESCC. Immunofluorescence was used to analyze fucosylation level. N-glycoproteomics and proteomics analysis, Lens Culinaris Agglutinin (LCA) and Ulex Europaeus Agglutinin I (UEA-I) affinity chromatography, immunoprecipitation, glycosyltransferase activity kit and rescue assay were used to explore the mechanism of TSTA3./nResults: TSTA3 was frequently amplified and overexpressed in ESCC. TSTA3 amplification and protein overexpression were significantly associated with malignant progression and poor prognosis of ESCC patients. TSTA3 knockdown significantly suppressed ESCC cells invasion and tumor dissemination by decreasing fucosylation level. Conversely, exogenous overexpression of TSTA3 led to increased invasion and tumor metastasis in vitro and in vivo by increasing fucosylation level. Moreover, core fucosylated LAMP2 and terminal fucosylated ERBB2 might be mediators of TSTA3-induced pro-invasion in ESCC and had a synergistic effect on the process. Peracetylated 2-F-Fuc, a fucosyltransferase activity inhibitor, reduced TSTA3 expression and fucosylation modification of LAMP2 and ERBB2, thereby inhibiting ESCC cell invasion./nConclusion: Our results indicate that TSTA3 may be a driver of ESCC metastasis through regulating fucosylation of LAMP2 and ERBB2. Fucosylation inhibitor may have prospect to suppress ESCC metastasis by blocking aberrant fucosylation.

中文翻译：

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TSTA3通过调节LAMP2和ERBB2的岩藻糖基化促进食管鳞状细胞癌的发展

背景： TSTA3基因编码一种酶，负责合成岩藻糖基化反应中的唯一供体GDP-L-岩藻糖。本研究旨在探讨TSTA3在食管鳞状细胞癌（ESCC）发生中的临床价值，功能和潜在机制。/n方法：将663位ESCC患者的全基因组测序数据和155位ESCC患者的RNA测序数据用于分别分析TSTA3的拷贝数变异和mRNA表达。基于或不基于组织微阵列的免疫组织化学用于检测其蛋白质表达。Transwell分析和体内转移试验用于研究TSTA3对ESCC侵袭和转移的影响。免疫荧光用于分析岩藻糖基化水平。的N- glycoproteomics和蛋白质组学分析，扁豆凝集素（LCA）和荆豆凝集素I（UEA-I）亲和层析，免疫沉淀，糖基转移酶活试剂盒和救援测定中用于探索TSTA3./n的机制结果：TSTA3在ESCC中经常被扩增和过表达。TSTA3扩增和蛋白过表达与ESCC患者的恶性进展和不良预后显着相关。通过降低岩藻糖基化水平，TSTA3敲低显着抑制了ESCC细胞的侵袭和肿瘤扩散。相反地，TSTA3的外源性过表达导致增加的侵袭和肿瘤转移在体外和体内通过增加岩藻糖基化水平。此外，核心岩藻糖基化的LAMP2和末端岩藻糖基化的ERBB2可能是TSTA3诱导的ESCC侵袭前体的介质，并对该过程具有协同作用。过乙酰化的2-F-Fuc，一种岩藻糖基转移酶活性抑制剂，可降低TSTA3的表达并减少LAMP2和ERBB2的岩藻糖基化修饰，从而抑制ESCC细胞侵袭。/n结论：我们的结果表明，TSTA3可能通过调节LAMP2的岩藻糖基化而成为ESCC转移的驱动器。和ERBB2。岩藻糖基化抑制剂有望通过阻断异常的岩藻糖基化来抑制ESCC转移。