Table1_Heparanase 1 Upregulation Promotes Tumor Progression and Is a Predictor of Low Survival for Oral Cancer.docx

Background: Oral cavity cancer is still an important public health problem throughout the world. Oral squamous cell carcinomas (OSCCs) can be quite aggressive and metastatic, with a low survival rate and poor prognosis. However, this is usually related to the clinical stage and histological grade, and molecular prognostic markers for clinical practice are yet to be defined. Heparanase (HPSE1) is an endoglycosidase associated with extracellular matrix remodeling, and although involved in several malignancies, the clinical implications of HPSE1 expression in OSCCs are still unknown.
Methods: We sought to investigate HPSE1 expression in a series of primary OSCCs and further explore whether its overexpression plays a relevant role in OSCC tumorigenesis. mRNA and protein expression analyses were performed in OSCC tissue samples and cell lines. A loss-of-function strategy using shRNA and a gain-of-function strategy using an ORF vector targeting HPSE1 were employed to investigate the endogenous modulation of HPSE1 and its effects on proliferation, apoptosis, adhesion, epithelial–mesenchymal transition (EMT), angiogenesis, migration, and invasion of oral cancer in vitro.
Results: We demonstrated that HPSE1 is frequently upregulated in OSCC samples and cell lines and is an unfavorable prognostic indicator of disease-specific survival when combined with advanced pT stages. Moreover, abrogation of HPSE1 in OSCC cells significantly promoted apoptosis and inhibited proliferation, migration, invasion, and epithelial–mesenchymal transition by significantly decreasing the expression of N-cadherin and vimentin. Furthermore, a conditioned medium of HPSE1-downregulated cells resulted in reduced vascular endothelial growth.
Conclusion: Our results confirm the overexpression of HPSE1 in OSCCs, suggest that HPSE1 expression correlates with disease progression as it is associated with several important biological processes for oral tumorigenesis, and can be managed as a prognostic marker for patients with OSCC.

Background: 口腔癌仍是全球范围内亟待解决的重要公共卫生问题。口腔鳞状细胞癌（Oral Squamous Cell Carcinomas, OSCCs）侵袭性强且易发生转移，患者生存率低下且预后不佳。不过这类不良预后通常与临床分期及组织学分级相关，目前临床仍缺乏明确可用的分子预后标志物。乙酰肝素酶（Heparanase, HPSE1）是一种参与细胞外基质重塑的内切糖苷酶，尽管其已被证实与多种恶性肿瘤相关，但HPSE1在OSCC中的表达及其临床意义仍未明确。
Methods: 本研究旨在探究一系列原发性OSCC样本中HPSE1的表达情况，并进一步明确其过表达是否在OSCC肿瘤发生过程中发挥关键作用。研究人员对OSCC组织样本及细胞系开展了mRNA与蛋白表达分析。实验采用靶向HPSE1的短发夹RNA（short hairpin RNA, shRNA）构建功能缺失模型，同时利用开放阅读框（Open Reading Frame, ORF）载体构建功能获得模型，以此探究HPSE1的内源性调控机制，并分析其对口腔癌细胞增殖、凋亡、黏附、上皮间质转化（Epithelial-Mesenchymal Transition, EMT）、血管生成、迁移及侵袭的体外影响。
Results: 本研究证实，HPSE1在OSCC组织样本及细胞系中普遍呈高表达；当其与进展性pT分期联合评估时，HPSE1高表达是患者疾病特异性生存率降低的不良预后因素。此外，在OSCC细胞中敲低HPSE1的表达可显著促进细胞凋亡，并抑制增殖、迁移、侵袭及上皮间质转化过程，其机制与显著下调N-钙黏蛋白（N-cadherin）与波形蛋白（Vimentin）的表达密切相关。进一步研究发现，HPSE1低表达细胞的条件培养基可减弱血管内皮生长效应。
Conclusion: 本研究结果证实了HPSE1在OSCC中存在过表达；HPSE1的表达与疾病进展显著相关，因其参与口腔肿瘤发生过程中的多项关键生物学进程，有望作为OSCC患者的临床预后标志物。