【摘要】：目的:Nemo樣激酶(nemo-like kinase,NLK)的異常表達(dá)在人類腫瘤的發(fā)生、侵襲和遷移中的作用已經(jīng)得以證實(shí)。然而,NLK在口腔鱗狀細(xì)胞癌(oral squamous cell carcinoma,OSCC,以下簡(jiǎn)稱口腔鱗癌)中的表達(dá)狀況以及可能發(fā)揮的作用仍未可知。本研究旨在探索NLK在口腔鱗癌中的表達(dá)水平,探討NLK與口腔鱗癌侵襲轉(zhuǎn)移的關(guān)系及其潛在機(jī)制。材料和方法:1、收集天津醫(yī)科大學(xué)附屬腫瘤醫(yī)院頜面耳鼻喉腫瘤科2006年1月至2010年10月口腔鱗癌手術(shù)標(biāo)本共計(jì)124例。另取43例口腔鱗癌癌旁組織作為對(duì)照。相關(guān)臨床病歷資料均完整,124例口腔鱗癌患者的生存數(shù)據(jù)由電話隨訪獲得。免疫組化實(shí)驗(yàn),檢測(cè)NLK、β-catenin、上皮間質(zhì)轉(zhuǎn)化(epithelial mesenchymal transition,EMT)相關(guān)蛋白及基質(zhì)金屬蛋白酶-2/9(matrix metalloproteinase-2/9,MMP-2/9)在口腔鱗癌中的表達(dá)情況,分析NLK與β-catenin、腫瘤臨床病理特征、患者預(yù)后和EMT的相關(guān)性。2、采用lipofectamine2000介導(dǎo)NLK si NLK轉(zhuǎn)染口腔鱗癌細(xì)胞SCC25和UM1,敲低NLK表達(dá),通過Western blot實(shí)驗(yàn)檢測(cè)β-catenin、EMT相關(guān)蛋白和MMP-2/9的表達(dá)情況。劃痕和transwell實(shí)驗(yàn)檢測(cè)口腔鱗癌細(xì)胞的遷移和侵襲能力變化。結(jié)果:1、124例口腔鱗癌中,82例(66.1%)NLK過表達(dá),顯著高于癌旁組織的16例(37.2%)。相較于非淋巴結(jié)轉(zhuǎn)移組,淋巴結(jié)轉(zhuǎn)移組的口腔鱗癌組織NLK陽性率更高,且中低分化組NLK表達(dá)明顯高于高分化組。另外,口腔鱗癌組織中,β-catenin和EMT相關(guān)表征蛋白(N-cadherin、Vimentin)以及侵襲指標(biāo)(MMP-2、MMP-9)過表達(dá),而E-cadherin低表達(dá)。統(tǒng)計(jì)分析發(fā)現(xiàn),NLK、β-catenin、EMT相關(guān)蛋白和侵襲相關(guān)指標(biāo)的表達(dá)與淋巴結(jié)轉(zhuǎn)移顯著相關(guān);NLK表達(dá)與β-catenin、N-cadherin、Vimentin、MMP-2和MMP-9表達(dá)呈正相關(guān),而與E-cadherin表達(dá)呈負(fù)相關(guān)。Cox多因素分析表明,NLK過表達(dá)、組織中低分化和淋巴結(jié)轉(zhuǎn)移是口腔鱗癌患者的相對(duì)獨(dú)立預(yù)后因素。Kaplan-Meier生存曲線顯示高表達(dá)NLK患者預(yù)后較差。2、敲低人口腔鱗癌SCC25和UM1細(xì)胞NLK表達(dá)發(fā)現(xiàn),β-catenin和p-β-catenin表達(dá)降低,且β-catenin細(xì)胞核中降低更為顯著。3、EMT相關(guān)蛋白(N-cadherin、Vimentin)和侵襲指標(biāo)(MMP-2、MMP-9)表達(dá)降低,而E-cadherin表達(dá)升高;此外,敲低NLK表達(dá),口腔鱗癌細(xì)胞的遷移和侵襲能力受到抑制。結(jié)論:1、NLK在口腔鱗癌中過表達(dá),與腫瘤病理分級(jí)、淋巴結(jié)轉(zhuǎn)移、患者預(yù)后和EMT顯著相關(guān)。2、NLK參與調(diào)控口腔鱗癌EMT,促進(jìn)口腔鱗癌侵襲轉(zhuǎn)移,這一調(diào)控作用可能是通過β-catenin通路實(shí)現(xiàn)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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R739.8

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本文編號(hào)：2396884