喉鳞癌中Twist、E-cadherin和N-cadherin基因表达及相关研究

发布时间：2019-05-22 17:49

【摘要】： 目的:喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)是耳鼻咽喉-头颈外科常见的恶性肿瘤之一,近年来发病率有日益增多的趋势。LSCC的发生和其他恶性肿瘤一样,可能和多因素引起相关基因表达水平的改变有关。扭曲基因(Twist)蛋白是属于碱性的螺旋-环-螺旋蛋白家族中的高度保守的转录因子,在胚胎的发生发展中发挥重要作用。Twist作为上皮-间质转变(Epithelial-mesenchymal transition,EMT)过程中的关键调控因子,对肿瘤的侵袭和转移有重要影响。本研究探讨了喉鳞癌组织中扭曲基因(Twist)、E-钙粘附素(E-cadherin)和N-钙粘附素(N-cadherin)的表达,进一步了解三者在喉鳞癌发生、发展、浸润、转移中的作用及三者之间的相互关系。 方法: 1应用免疫组织化学(immunohistochemistry, IHC)法检测49例喉鳞癌及20例癌旁组织中Twist、E-cadherin和N-cadherin基因的蛋白表达情况。 2应用逆转录-多聚酶链反应(RT-PCR)检测49例喉鳞癌及20例癌旁组织中Twist、E-cadherin和N-cadherin的mRNA表达情况。 3统计学处理:应用SPSS13.0统计软件对数据进行统计分析。以P≤0.05为差异在统计学上有显著意义,以P≤0.01为差异在统计学上有非常显著意义 结果: 1 Twist的表达及其与临床参数之间的关系 1.1:应用免疫组化法检测49例喉鳞癌组织和20例癌旁组织中Twist蛋白表达情况,阳性表达率分别为61.22%(30/49)和25.00%(5/20),二者差异统计学上具有非常显著性意义(P0.01),Twist在喉鳞癌组织中的蛋白表达明显高于癌旁组织。在喉鳞癌中Twist的蛋白表达与淋巴结转移(N+组高于N0组,P0.05)、临床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)相关,而与肿瘤病理学分级、临床分型及年龄分组无关(P0.05)。 1.2应用RT-PCR法检测49例喉鳞癌组织和20例癌旁组织中Twist mRNA的表达情况,其相对表达量分别为(0.93±0.39)和(0.57±0.24),两者差异在统计学上有非常显著性意义(P0.01),Twist在喉鳞癌组织中的mRNA表达明显高于癌旁组织;在喉鳞癌中Twist mRNA的表达与淋巴结转移(N+组高于N0组,P0.05)、临床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)相关,而与肿瘤病理学分级、临床分型及年龄分组无关(P0.05)。 2 E-cadherin的表达及其与临床参数之间的关系 2.1应用免疫组化法检测49例喉鳞癌组织和20例癌旁组织中的E-cadherin蛋白表达情况,阳性表达率分别为51.02%(25/49)和100.00%(20/20),二者差异在统计学上有非常显著性意义(P0.01),E-cadherin在喉鳞癌组织中的蛋白表达明显低于癌旁组织;在喉癌中E-cadherin的表达与肿瘤病理学分级(G1、G2组高于G3组,P0.05)、淋巴结转移(N0组高于N+组,P0.01)和临床分期(Ⅰ、Ⅱ期高于Ⅲ、Ⅳ期,P0.01)相关,而与临床分型及年龄分组无关(P0.05)。 2.2应用RT-PCR法检测49例喉鳞癌组织和20例癌旁组织中的E-cadherin mRNA的表达情况,其相对表达量分别为(0.64±0.30)和(0.93±0.33),前者明显低于后者,二者差异在统计学上有非常显著性意义(P0.01)。E-cadherin在喉鳞癌组织中的mRNA表达明显低于癌旁组织;在喉鳞癌中E-cadherin的mRNA表达与肿瘤病理学分级(G1、G2组高于G3组,P0.01)、淋巴结转移(N0组高于N+组,P0.01)和临床分期(Ⅰ、Ⅱ期高于Ⅲ、Ⅳ期,P0.05)相关,而与临床分型及年龄分组无关(P0.05)。 3 N-cadherin的表达及其与临床参数之间的关系 3.1应用免疫组化法检测49例喉鳞癌组织和20例癌旁组织中的N-cadherin蛋白表达情况,阳性表达率分别为73.47%(36/49)和40.00%(8/20),二者差异具有显著性意义(P0.05),N-cadherin在喉鳞癌组织中的蛋白表达明显高于癌旁组织;在喉鳞癌中N-cadherin蛋白表达与淋巴结转移(N+组高于N0组,P0.05)、临床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)及年龄分组(≤60岁组高于60岁组,P0.05)相关,而与肿瘤病理学分级及临床分型分组无关(P0.05)。 3.2应用RT-PCR法检测49例喉鳞癌组织和20例癌旁组织中的N-cadherin mRNA的表达情况,其相对表达量分别为(0.70±0.21)和(0.36±0.11),二者差异在统计学上有非常显著性意义(P0.01)。N-cadherin在喉鳞癌组织中的mRNA表达明显高于癌旁组织;在喉鳞癌中N-cadherin的mRNA表达与淋巴结转移(N+组高于N0组,P0.05)、临床分期(Ⅲ、Ⅳ期高于Ⅰ、Ⅱ期,P0.01)相关,而与肿瘤病理学分级、临床分型、年龄分组无关(P0.05)。 4三种基因mRNA及蛋白表达相关性:在mRNA水平及蛋白水平,Twist与E-cadherin (r=-0.760,P0.01;r=-0.662,P0.01),N-cadherin与E-cadherin (r=-0.615,P0.01;r=-0.558,P0.01)的表达均呈负相关;Twist与N-cadherin均呈正相关(r=0.850,P0.01;r=0.649,P0.01)。 结论: 1喉鳞癌中,Twist和N-cadherin基因mRNA及蛋白水平均呈高表达,E-cadherin均呈低表达,提示三者可能与喉癌的发生相关。 2喉鳞癌中Twist和N-cadherin的高表达和E-cadherin的低表达与淋巴结转移、临床分期等喉癌恶性生物学行为相关,提示三者可能参与了喉癌的浸润、转移。 3喉鳞癌中Twist和N-cadherin的表达呈正相关,和E-cadherin的表达呈负相关,提示Twist可能通过抑制E-cadherin,促进N-cadherin的表达,从而参与喉癌的浸润和转移。
[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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