Semaphorin7A Promotion of Tumoral Growth and Metastasis in Human Oral Cancer by Regulation of G1 Cell Cycle and Matrix Metalloproteases: Possible Contribution to Tumoral Angiogenesis.
Semaphorin7A Promotion of Tumoral Growth and Metastasis in Human Oral Cancer by Regulation of G1 Cell Cycle and Matrix Metalloproteases: Possible Contribution to Tumoral Angiogenesis.
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DOI：
10.1371/journal.pone.0137923
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Uzawa K
中科院分区：
文献类型：
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作者：
Saito T;Kasamatsu A;Ogawara K;Miyamoto I;Saito K;Iyoda M;Suzuki T;Endo-Sakamoto Y;Shiiba M;Tanzawa H;Uzawa K
信号蛋白（SEMAs）由一个大的分泌和膜锚定蛋白家族组成，在发育中的神经系统的选定区域的神经元寻路和轴突引导中起重要作用。其中，SEMA7A已被报道在神经发生中具有趋化活性和免疫调节剂；然而，SEMA7A与口腔鳞状细胞癌（OSCC）行为的相关性知之甚少。我们使用定量逆转录聚合酶链反应、免疫印迹和半定量免疫组织化学（sq-IHC）评估了SEMA7A在OSCC衍生细胞系和原代OSCC样本中的表达。此外，SEMA7A敲低细胞（shSEMA7A细胞）被用于功能实验，包括细胞增殖、侵袭性和迁移测定。我们还分析了OSCC患者SEMA7A状态与临床行为的临床相关性。与人正常口腔角质形成细胞相比，oscc来源细胞系中SEMA7A mRNA和蛋白表达显著上调（P<0.05）。shSEMA7A细胞在G1期表现出细胞周期阻滞，细胞周期依赖性激酶抑制剂（p21Cip1和p27Kip1）上调，细胞周期蛋白（cyclin D1, cyclin E）和细胞周期蛋白依赖性激酶（CDK2， CDK4和CDK6）下调；基质金属蛋白酶（MMPs） （MMP-2、proMMP-2、pro-MMP-9）的分泌减少，膜型1-MMP （MT1-MMP）的表达减少，从而降低侵袭性和迁移活性。我们还发现细胞外调节的激酶1/2和AKT通路失活，这是细胞周期阻滞在G1期的上游分子，并且减少了shSEMA7A细胞中MMPs的分泌。sq-IHC结果显示，SEMA7A在原发OSCCs中的表达显著高于正常OSCCs (P = 0.001)，且与原发肿瘤大小（P = 0.0254）和区域淋巴结转移（P = 0.0002）相关。我们的数据为SEMA7A在OSCC的肿瘤生长和转移中发挥重要作用提供了证据，并表明SEMA7A可能在OSCC患者中发挥潜在的诊断/治疗靶点。
Semaphorins (SEMAs) consist of a large family of secreted and membrane-anchored proteins that are important in neuronal pathfinding and axon guidance in selected areas of the developing nervous system. Of them, SEMA7A has been reported to have a chemotactic activity in neurogenesis and to be an immunomodulator; however, little is known about the relevance of SEMA7A in the behaviors of oral squamous cell carcinoma (OSCC). We evaluated SEMA7A expression in OSCC-derived cell lines and primary OSCC samples using quantitative reverse transcriptase-polymerase chain reaction, immunoblotting, and semiquantitative immunohistochemistry (sq-IHC). In addition, SEMA7A knockdown cells (shSEMA7A cells) were used for functional experiments, including cellular proliferation, invasiveness, and migration assays. We also analyzed the clinical correlation between SEMA7A status and clinical behaviors in patients with OSCC. SEMA7A mRNA and protein were up-regulated significantly (P<0.05) in OSCC-derived cell lines compared with human normal oral keratinocytes. The shSEMA7A cells showed decreased cellular growth by cell-cycle arrest at the G1 phase, resulting from up-regulation of cyclin-dependent kinase inhibitors (p21Cip1 and p27Kip1) and down-regulation of cyclins (cyclin D1, cyclin E) and cyclin-dependent kinases (CDK2, CDK4, and CDK6); and decreased invasiveness and migration activities by reduced secretion of matrix metalloproteases (MMPs) (MMP-2, proMMP-2, pro-MMP-9), and expression of membrane type 1- MMP (MT1-MMP). We also found inactivation of the extracellular regulated kinase 1/2 and AKT pathways, an upstream molecule of cell-cycle arrest at the G1 phase, and reduced secretion of MMPs in shSEMA7A cells. sq-IHC showed that SEMA7A expression in the primary OSCCs was significantly (P = 0.001) greater than that in normal counterparts and was correlated with primary tumoral size (P = 0.0254) and regional lymph node metastasis (P = 0.0002). Our data provide evidence for an essential role of SEMA7A in tumoral growth and metastasis in OSCC and indicated that SEMA7A may play a potential diagnostic/therapeutic target for use in patients with OSCC.