NLK通过β-catenin通路促进口腔鳞癌侵袭转移的研究

发布时间：2018-12-31 17:25

【摘要】：目的:Nemo样激酶(nemo-like kinase,NLK)的异常表达在人类肿瘤的发生、侵袭和迁移中的作用已经得以证实。然而,NLK在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC,以下简称口腔鳞癌)中的表达状况以及可能发挥的作用仍未可知。本研究旨在探索NLK在口腔鳞癌中的表达水平,探讨NLK与口腔鳞癌侵袭转移的关系及其潜在机制。材料和方法:1、收集天津医科大学附属肿瘤医院颌面耳鼻喉肿瘤科2006年1月至2010年10月口腔鳞癌手术标本共计124例。另取43例口腔鳞癌癌旁组织作为对照。相关临床病历资料均完整,124例口腔鳞癌患者的生存数据由电话随访获得。免疫组化实验,检测NLK、β-catenin、上皮间质转化(epithelial mesenchymal transition,EMT)相关蛋白及基质金属蛋白酶-2/9(matrix metalloproteinase-2/9,MMP-2/9)在口腔鳞癌中的表达情况,分析NLK与β-catenin、肿瘤临床病理特征、患者预后和EMT的相关性。2、采用lipofectamine2000介导NLK si NLK转染口腔鳞癌细胞SCC25和UM1,敲低NLK表达,通过Western blot实验检测β-catenin、EMT相关蛋白和MMP-2/9的表达情况。划痕和transwell实验检测口腔鳞癌细胞的迁移和侵袭能力变化。结果:1、124例口腔鳞癌中,82例(66.1%)NLK过表达,显著高于癌旁组织的16例(37.2%)。相较于非淋巴结转移组,淋巴结转移组的口腔鳞癌组织NLK阳性率更高,且中低分化组NLK表达明显高于高分化组。另外,口腔鳞癌组织中,β-catenin和EMT相关表征蛋白(N-cadherin、Vimentin)以及侵袭指标(MMP-2、MMP-9)过表达,而E-cadherin低表达。统计分析发现,NLK、β-catenin、EMT相关蛋白和侵袭相关指标的表达与淋巴结转移显著相关;NLK表达与β-catenin、N-cadherin、Vimentin、MMP-2和MMP-9表达呈正相关,而与E-cadherin表达呈负相关。Cox多因素分析表明,NLK过表达、组织中低分化和淋巴结转移是口腔鳞癌患者的相对独立预后因素。Kaplan-Meier生存曲线显示高表达NLK患者预后较差。2、敲低人口腔鳞癌SCC25和UM1细胞NLK表达发现,β-catenin和p-β-catenin表达降低,且β-catenin细胞核中降低更为显著。3、EMT相关蛋白(N-cadherin、Vimentin)和侵袭指标(MMP-2、MMP-9)表达降低,而E-cadherin表达升高;此外,敲低NLK表达,口腔鳞癌细胞的迁移和侵袭能力受到抑制。结论:1、NLK在口腔鳞癌中过表达,与肿瘤病理分级、淋巴结转移、患者预后和EMT显著相关。2、NLK参与调控口腔鳞癌EMT,促进口腔鳞癌侵袭转移,这一调控作用可能是通过β-catenin通路实现的。
[Abstract]:Objective: the abnormal expression of Nemo-like kinase (nemo-like kinase,NLK) has been demonstrated in the pathogenesis, invasion and migration of human tumors. However, the expression and possible role of NLK in oral squamous cell carcinoma (oral squamous cell carcinoma,OSCC,) remains unknown. The purpose of this study was to investigate the expression of NLK in oral squamous cell carcinoma (OSCC) and to explore the relationship between NLK and invasion and metastasis of OSCC and its underlying mechanism. Materials and methods: 1. 124 cases of oral squamous cell carcinoma were collected from Department of Otolaryngology, affiliated Cancer Hospital of Tianjin Medical University from January 2006 to October 2010. Another 43 cases of oral squamous cell carcinoma adjacent tissues were used as control. The survival data of 124 patients with oral squamous cell carcinoma were obtained by telephone follow-up. Immunohistochemical method was used to detect the expression of NLK, 尾-catenin, (epithelial mesenchymal transition,EMT) related protein and matrix metalloproteinase-2 / 9 (matrix metalloproteinase-2/9,MMP-2/9) in oral squamous cell carcinoma (OSCC). The clinicopathological features of NLK and 尾-catenin, tumors, the relationship between prognosis and EMT were analyzed. 2. Lipofectamine2000 mediated NLK si NLK was used to transfect SCC25 and UM1, knockout NLK expression in oral squamous carcinoma cells, and 尾-catenin, was detected by Western blot assay. Expression of EMT related proteins and MMP-2/9. The migration and invasiveness of oral squamous carcinoma cells were detected by scratch and transwell assay. Results: in 1124 cases of oral squamous cell carcinoma, 82 cases (66.1%) were overexpression of NLK, which was significantly higher than that in 16 cases (37.2%) of adjacent tissues. Compared with the non-lymph node metastasis group, the positive rate of NLK was higher in the lymph node metastasis group than in the non-lymph node metastasis group, and the expression of NLK in the middle and low differentiation group was significantly higher than that in the well-differentiated group. In addition, 尾-catenin and EMT related protein (N-cadherin vimentin) and invasion index (MMP-2,MMP-9) were overexpressed in oral squamous cell carcinoma (OSCC), but E-cadherin was low expression. Statistical analysis showed that the expression of NLK, 尾-catenin,EMT related protein and invasion-related markers were significantly correlated with lymph node metastasis. The expression of NLK was positively correlated with the expression of 尾-catenin,N-cadherin,Vimentin,MMP-2 and MMP-9, but negatively correlated with the expression of E-cadherin. Cox multivariate analysis showed that NLK was overexpressed. Low differentiation and lymph node metastasis were relative independent prognostic factors in patients with oral squamous cell carcinoma (OSCC). Kaplan-Meier survival curve showed poor prognosis in patients with high expression of NLK. The expression of 尾-catenin and p- 尾-catenin was decreased, especially in the nucleus of 尾-catenin. 3The expression of N-cadherin associated protein (N-cadherin Vimentin) and invasion index (MMP-2,MMP-9) decreased, while the expression of E-cadherin increased. In addition, knockout NLK expression inhibited the migration and invasion of oral squamous carcinoma cells. Conclusion: 1the overexpression of NLK in oral squamous cell carcinoma is significantly correlated with tumor pathological grade, lymph node metastasis, prognosis and EMT. 2NLK is involved in the regulation of EMT, in oral squamous cell carcinoma to promote invasion and metastasis of oral squamous cell carcinoma. This regulation may be achieved through the 尾-catenin pathway.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8

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