【摘要】： 目的:喉鱗狀細胞癌(laryngeal squamous cell carcinoma,LSCC)是耳鼻咽喉-頭頸外科常見的惡性腫瘤之一,近年來發(fā)病率有日益增多的趨勢。LSCC的發(fā)生和其他惡性腫瘤一樣,可能和多因素引起相關(guān)基因表達水平的改變有關(guān)。扭曲基因(Twist)蛋白是屬于堿性的螺旋-環(huán)-螺旋蛋白家族中的高度保守的轉(zhuǎn)錄因子,在胚胎的發(fā)生發(fā)展中發(fā)揮重要作用。Twist作為上皮-間質(zhì)轉(zhuǎn)變(Epithelial-mesenchymal transition,EMT)過程中的關(guān)鍵調(diào)控因子,對腫瘤的侵襲和轉(zhuǎn)移有重要影響。本研究探討了喉鱗癌組織中扭曲基因(Twist)、E-鈣粘附素(E-cadherin)和N-鈣粘附素(N-cadherin)的表達,進一步了解三者在喉鱗癌發(fā)生、發(fā)展、浸潤、轉(zhuǎn)移中的作用及三者之間的相互關(guān)系。 方法: 1應用免疫組織化學(immunohistochemistry, IHC)法檢測49例喉鱗癌及20例癌旁組織中Twist、E-cadherin和N-cadherin基因的蛋白表達情況。 2應用逆轉(zhuǎn)錄-多聚酶鏈反應(RT-PCR)檢測49例喉鱗癌及20例癌旁組織中Twist、E-cadherin和N-cadherin的mRNA表達情況。 3統(tǒng)計學處理:應用SPSS13.0統(tǒng)計軟件對數(shù)據(jù)進行統(tǒng)計分析。以P≤0.05為差異在統(tǒng)計學上有顯著意義,以P≤0.01為差異在統(tǒng)計學上有非常顯著意義 結(jié)果: 1 Twist的表達及其與臨床參數(shù)之間的關(guān)系 1.1:應用免疫組化法檢測49例喉鱗癌組織和20例癌旁組織中Twist蛋白表達情況,陽性表達率分別為61.22%(30/49)和25.00%(5/20),二者差異統(tǒng)計學上具有非常顯著性意義(P0.01),Twist在喉鱗癌組織中的蛋白表達明顯高于癌旁組織。在喉鱗癌中Twist的蛋白表達與淋巴結(jié)轉(zhuǎn)移(N+組高于N0組,P0.05)、臨床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)相關(guān),而與腫瘤病理學分級、臨床分型及年齡分組無關(guān)(P0.05)。 1.2應用RT-PCR法檢測49例喉鱗癌組織和20例癌旁組織中Twist mRNA的表達情況,其相對表達量分別為(0.93±0.39)和(0.57±0.24),兩者差異在統(tǒng)計學上有非常顯著性意義(P0.01),Twist在喉鱗癌組織中的mRNA表達明顯高于癌旁組織;在喉鱗癌中Twist mRNA的表達與淋巴結(jié)轉(zhuǎn)移(N+組高于N0組,P0.05)、臨床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)相關(guān),而與腫瘤病理學分級、臨床分型及年齡分組無關(guān)(P0.05)。 2 E-cadherin的表達及其與臨床參數(shù)之間的關(guān)系 2.1應用免疫組化法檢測49例喉鱗癌組織和20例癌旁組織中的E-cadherin蛋白表達情況,陽性表達率分別為51.02%(25/49)和100.00%(20/20),二者差異在統(tǒng)計學上有非常顯著性意義(P0.01),E-cadherin在喉鱗癌組織中的蛋白表達明顯低于癌旁組織;在喉癌中E-cadherin的表達與腫瘤病理學分級(G1、G2組高于G3組,P0.05)、淋巴結(jié)轉(zhuǎn)移(N0組高于N+組,P0.01)和臨床分期(Ⅰ、Ⅱ期高于Ⅲ、Ⅳ期,P0.01)相關(guān),而與臨床分型及年齡分組無關(guān)(P0.05)。 2.2應用RT-PCR法檢測49例喉鱗癌組織和20例癌旁組織中的E-cadherin mRNA的表達情況,其相對表達量分別為(0.64±0.30)和(0.93±0.33),前者明顯低于后者,二者差異在統(tǒng)計學上有非常顯著性意義(P0.01)。E-cadherin在喉鱗癌組織中的mRNA表達明顯低于癌旁組織;在喉鱗癌中E-cadherin的mRNA表達與腫瘤病理學分級(G1、G2組高于G3組,P0.01)、淋巴結(jié)轉(zhuǎn)移(N0組高于N+組,P0.01)和臨床分期(Ⅰ、Ⅱ期高于Ⅲ、Ⅳ期,P0.05)相關(guān),而與臨床分型及年齡分組無關(guān)(P0.05)。 3 N-cadherin的表達及其與臨床參數(shù)之間的關(guān)系 3.1應用免疫組化法檢測49例喉鱗癌組織和20例癌旁組織中的N-cadherin蛋白表達情況,陽性表達率分別為73.47%(36/49)和40.00%(8/20),二者差異具有顯著性意義(P0.05),N-cadherin在喉鱗癌組織中的蛋白表達明顯高于癌旁組織;在喉鱗癌中N-cadherin蛋白表達與淋巴結(jié)轉(zhuǎn)移(N+組高于N0組,P0.05)、臨床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)及年齡分組(≤60歲組高于60歲組,P0.05)相關(guān),而與腫瘤病理學分級及臨床分型分組無關(guān)(P0.05)。 3.2應用RT-PCR法檢測49例喉鱗癌組織和20例癌旁組織中的N-cadherin mRNA的表達情況,其相對表達量分別為(0.70±0.21)和(0.36±0.11),二者差異在統(tǒng)計學上有非常顯著性意義(P0.01)。N-cadherin在喉鱗癌組織中的mRNA表達明顯高于癌旁組織;在喉鱗癌中N-cadherin的mRNA表達與淋巴結(jié)轉(zhuǎn)移(N+組高于N0組,P0.05)、臨床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)相關(guān),而與腫瘤病理學分級、臨床分型、年齡分組無關(guān)(P0.05)。 4三種基因mRNA及蛋白表達相關(guān)性:在mRNA水平及蛋白水平,Twist與E-cadherin (r=-0.760,P0.01;r=-0.662,P0.01),N-cadherin與E-cadherin (r=-0.615,P0.01;r=-0.558,P0.01)的表達均呈負相關(guān);Twist與N-cadherin均呈正相關(guān)(r=0.850,P0.01;r=0.649,P0.01)。 結(jié)論: 1喉鱗癌中,Twist和N-cadherin基因mRNA及蛋白水平均呈高表達,E-cadherin均呈低表達,提示三者可能與喉癌的發(fā)生相關(guān)。 2喉鱗癌中Twist和N-cadherin的高表達和E-cadherin的低表達與淋巴結(jié)轉(zhuǎn)移、臨床分期等喉癌惡性生物學行為相關(guān),提示三者可能參與了喉癌的浸潤、轉(zhuǎn)移。 3喉鱗癌中Twist和N-cadherin的表達呈正相關(guān),和E-cadherin的表達呈負相關(guān),提示Twist可能通過抑制E-cadherin,促進N-cadherin的表達,從而參與喉癌的浸潤和轉(zhuǎn)移。
[Abstract]:Objective: Laryngeal square cell carcinoma (LSCC) is one of the most common malignant tumors of the otorhinolaryngology-head and neck surgery. The occurrence of LSCC, like other malignancies, may be related to a change in the level of gene expression associated with multiple factors. The Twist protein is a highly conserved transcription factor in the basic helical-loop-helical protein family and plays an important role in the development of the embryo. Twist is a key factor in the process of epithelial-mesenchymal transition (EMT), and has an important effect on the invasion and metastasis of the tumor. In this study, the expression of Twist, E-cadherin and N-cadherin in laryngeal squamous cell carcinoma was studied. Methods:1. The Twist, E-cadherin and N-cadherin genes in 49 cases of laryngeal squamous cell carcinoma and 20 other adjacent tissues were detected by immunohistochemistry (IHC). The expression of protein was detected by reverse transcription-polymerase chain reaction (RT-PCR). Twist, E-cadherin and N-cadherin in 49 cases of laryngeal squamous cell carcinoma and 20 cases of adjacent tissues were detected by reverse transcription-polymerase chain reaction (RT-PCR). mRNA expression of in. Statistical treatment: application of SPSS13 .0 Statistical software was used for statistical analysis of the data. The difference of P-0.05 was statistically significant, P <0.0 1 is poor The difference was statistically significant: 1 The expression of Twist and its relation to the clinical parameters were 1.1: The expression of Twist in 49 cases of laryngeal squamous cell carcinoma and 20 cases of adjacent tissues was detected by immunohistochemical method. The positive rate of expression was 61.22% (30/49) and 25.00% (5/20), respectively. The protein expression of Twist in laryngeal squamous cell carcinoma was significantly higher than that in the adjacent tissues. The expression of Twinist in laryngeal squamous cell carcinoma and lymph node metastasis (N + group was higher than that of N0 group, P0.05), and the clinical stage (stage 鈪，

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