Abstract：
Wiskott-Aldrich syndrome protein family verprolin-homologous domain-containing protein 3 (WAVE3) is reported as an oncogene regulating cell proliferation and motility in multiple malignancies, while its role in tongue squamous cell carcinoma (TSCC) remains unknown. This study aimed to explore the expression and mechanism of WAVE3 in TSCC. We enrolled 64 TSCC patients admitted between June 2013 and February 2014 and collected their cancerous and adjacent normal tissues to determine WAVE3 expression by immunohistochemistry. The correlation of WAVE3 expression with TSCC patients\' pathological characteristics was analyzed. Then, a 7-year follow-up was conducted to observe the value of WAVE3 in evaluating patient outcomes. In addition, human TSCC SCC9, SCC25, and CAL27 cells were purchased and detected by Cell Counting Kit-8 (CCK-8), Transwell, and scratch-wound assays for their proliferation, invasion, and migration capacities, while real-time quantitative PCR (qRT-PCR) and Western blotting were utilized to quantify WAVE3 and epithelial-mesenchymal transition (EMT)-related protein expression, respectively. The most active cell lines were selected to be infected with lentiviral vectors that silenced WAVE3 (named WAVE3-sh group) and overexpressed WAVE3 cDNA (named WAVE3-OE group) to observe the impacts of interfering WAVE3 expression on TSCC cell biological behavior. The positive expression of WAVE3 in TSCC tissue was found to be obviously enhanced and predominantly located in the cytoplasm. In addition, close correlations were identified between WAVE3 and T staging, clinical staging, lymphatic metastasis, distant metastasis, and differentiation degree (P < 0.05). Increased WAVE3 expression predicted an elevated risk of death, as indicated by the follow-up analysis (P < 0.05). SCC9 was selected for subsequent experiments among various TSCC cell lines studied because it showed the most potent ability to proliferate, invade, and migrate (P < 0.05). Silencing WAVE3 expression in SCC9 cells decreased cell proliferation, invasion, migration, and EMT-related protein expression (P < 0.05), while increasing WAVE3 expression promoted SCC9 viability. WAVE3, which was highly expressed in TSCC, promoted EMT in tumor cells and accelerated their proliferation, invasion, and migration, which might provide a new theoretical basis for molecular targeted therapy of TSCC in the future.
摘要：
据报道，Wiskott-Aldrich综合征蛋白家族含有verprolin同源域的蛋白3（WAVE3）是一种调节多种恶性肿瘤细胞增殖和运动的癌基因，而其在舌鳞状细胞癌（TSCC）中的作用尚不清楚。本研究旨在探讨WAVE3在TSCC中的表达及其作用机制。我们招募了2013年6月至2014年2月期间收治的64例TSCC患者，并收集其癌组织和邻近正常组织，以通过免疫组织化学确定WAVE3表达。分析WAVE3表达与TSCC患者病理特征的相关性。然后，我们进行了7年随访,观察WAVE3在评估患者结局方面的价值.此外,购买人TSCCSCC9，SCC25和CAL27细胞，并通过细胞计数试剂盒-8（CCK-8）检测，Transwell,以及它们增殖的划痕试验，入侵,和迁移能力，而实时定量PCR（qRT-PCR）和蛋白质印迹用于定量WAVE3和上皮间质转化（EMT）相关蛋白的表达，分别。选择最活跃的细胞系用沉默WAVE3（WAVE3-sh组）和过表达WAVE3cDNA（WAVE3-OE组）的慢病毒载体感染，观察干扰WAVE3表达对TSCC细胞生物学行为的影响。发现TSCC组织中WAVE3的阳性表达明显增强，并主要位于细胞质中。此外,确定WAVE3和T分期之间密切相关，临床分期,淋巴转移，远处转移,分化程度(P<0.05)。WAVE3表达增加预测死亡风险升高，如随访分析所示（P<0.05）。在研究的各种TSCC细胞系中选择SCC9进行后续实验，因为它显示出最有效的增殖能力，入侵,并迁移(P<0.05)。沉默WAVE3在SCC9细胞中的表达降低了细胞增殖,入侵,迁移,和EMT相关蛋白表达（P<0.05），而增加WAVE3表达促进SCC9活力。在TSCC中高表达的WAVE3，促进肿瘤细胞中的EMT并加速其增殖，入侵,和移民,为今后TSCC的分子靶向治疗提供新的理论依据。
