Rodrigues Priscila Campioni, Sawazaki-Calone Iris, Ervolino de Oliveira Carine, Soares Macedo Carolina Carneiro, Dourado Mauricio Rocha, Cervigne Nilva K, Miguel Marcia Costa, Ferreira do Carmo Andreia, Lambert Daniel W, Graner Edgard, Daniela da Silva Sabrina, Alaoui-Jamali Moulay A, Paes Leme Adriana Franco, Salo Tuula A, Coletta Ricardo D
Department of Oral Diagnosis, School of Dentistry, University of Campinas, Piracicaba, SP, Brazil.
Unit of Cancer Research and Translational Medicine, Faculty of Medicine and Medical Research Center Oulu, Oulu University Hospital, University of Oulu, Oulu, Finland.
Oncotarget. 2017 Aug 19;8(43):74736-74754. doi: 10.18632/oncotarget.20360. eCollection 2017 Sep 26.
Oral squamous cell carcinoma (OSCC) prognosis is related to clinical stage and histological grade. However, this stratification needs to be refined. We conducted a comparative proteome study in microdissected samples from normal oral mucosa and OSCC to identify biomarkers for malignancy. Fascin and plectin were identified as differently expressed and both are implicated in several malignancies, but the clinical impacts of aberrant fascin and plectin expression in OSCCs remains largely unknown. Immunohistochemistry and real-time quantitative PCR were carried out in OSCC samples and cell lines. A loss-of-function strategy using shRNA targeting fascin was employed to investigate and the fascin role on oral tumorigenesis. Transfections of microRNA mimics were performed to determine whether the fascin overexpression is regulated by miR-138 and miR-145. We found that fascin and plectin are frequently upregulated in OSCC samples and cell lines, but only fascin overexpression is an independent unfavorable prognostic indicator of disease-specific survival. In combination with advanced T stage, high fascin level is also an independent factor of disease-free survival. Knockdown of fascin in OSCC cells promoted cell adhesion and inhibited migration, invasion and EMT, and forced expression of miR-138 in OSCC cells significantly decreased the expression of fascin. In addition, fascin downregulation leads to reduced filopodia formation and decrease on paxillin expression. The subcutaneous xenograft model showed that tumors formed in the presence of low levels of fascin were significantly smaller compared to those formed with high fascin levels. Collectively, our findings suggest that fascin expression correlates with disease progression and may serve as a prognostic marker and therapeutic target for patients with OSCC.
口腔鳞状细胞癌（OSCC）的预后与临床分期和组织学分级相关。然而，这种分层需要进一步细化。我们对正常口腔黏膜和OSCC的显微切割样本进行了比较蛋白质组学研究，以确定恶性肿瘤的生物标志物。Fascin和盘状球蛋白被鉴定为表达差异，且二者均与多种恶性肿瘤有关，但Fascin和盘状球蛋白异常表达在OSCC中的临床影响仍 largely未知。对OSCC样本和细胞系进行了免疫组织化学和实时定量PCR检测。采用靶向Fascin的短发夹RNA（shRNA）的功能缺失策略来研究Fascin在口腔肿瘤发生中的作用。进行了微小RNA模拟物转染，以确定Fascin的过表达是否受miR - 138和miR - 145调控。我们发现，Fascin和盘状球蛋白在OSCC样本和细胞系中经常上调，但只有Fascin过表达是疾病特异性生存的独立不良预后指标。与晚期T分期相结合，高Fascin水平也是无病生存的独立因素。敲低OSCC细胞中的Fascin可促进细胞黏附，并抑制迁移、侵袭和上皮 - 间质转化（EMT），且在OSCC细胞中强制表达miR - 138可显著降低Fascin的表达。此外，Fascin下调导致丝状伪足形成减少和桩蛋白表达降低。皮下异种移植模型显示，与高Fascin水平形成的肿瘤相比，低Fascin水平下形成的肿瘤明显更小。总体而言，我们的研究结果表明，Fascin表达与疾病进展相关，可能作为OSCC患者的预后标志物和治疗靶点。
