OPN、CD44v6、MMP-9基因及其蛋白与喉鳞癌浸润转移关系的研究

发布时间：2018-01-16 10:26

本文关键词：OPN、CD44v6、MMP-9基因及其蛋白与喉鳞癌浸润转移关系的研究　出处：《河北北方学院》2012年硕士论文　论文类型：学位论文

【摘要】：本课题应用免疫组织化学技术(immunohistochemistry, IHC)和原位杂交技术(In situ hybridization,ISH)检测喉鳞状细胞癌(laryngeal squamous cell carcinoma, LSCC)组织和正常喉黏膜组织中骨桥蛋白(osteopontin, OPN)、细胞黏附分子CD44拼接变异体6(adhesion molecules CD44splice variant6, CD44v6)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的表达,探讨OPN、CD44v6、MMP-9mRNA及其蛋白在正常喉黏膜组织和喉鳞状细胞癌组织中的表达及与患者临床病理特征的关系；探讨OP、CD44v6、MMP-9mRNA及其蛋白在喉鳞状细胞癌组织中表达的相互关系。采用免疫组织化学技术和原位杂交技术方法检测47例喉鳞癌组织,10例正常的喉黏膜组织中OPN、CD44v6、MMP-9mRNA及其蛋白的表达情况；分析三个指标与患者的年龄、性别、分型、分化程度、TNM分期及淋巴结转移的关系以及三者之间表达的关系；分析两种方法检测OPN、CD44v6、MMP-9表达的一致性。 OPN、CD44v6、MMP-9蛋白在47例喉鳞癌组织中的阳性表达率分别为63.8%(30/47)、76.6%(36/47)、68.1%(32/47)；10例正常喉黏膜组织中的阳性率分别为：10%(1/10)、30%(3/10)、0%(0/10),喉鳞癌组织中OPN、CD44v6、MMP-9蛋白的阳性表达率明显高于正常喉黏膜组织中的表达率,差异显著(P0.05)。OPN、CD44v6蛋白在喉鳞癌组织中高表达与肿瘤分化程度、TNM分期、淋巴结转移相关,OPN、CD44v6蛋白与年龄、性别及肿瘤位置等临床病理学特征无明显关系；MMP-9蛋白在喉鳞癌组织中高表达与TNM分期、淋巴结转移相关,MMP-9蛋白与患者年龄、性别、肿瘤位置、分化程度无明显关系。在47例喉鳞癌组织标本中OPN、CD44v6、MMP-9蛋白阳性表达分别为30例、36例、32例,在OPN蛋白阳性表达30例中CD44v6蛋白阳性表达27例,MMP-9蛋白阳性表达24例,经Spearman相关分析,结果显示：OPN蛋白与CD44v6蛋白两者表达呈显著正相关；OPN蛋白与MMP-9蛋白两者表达呈显著正相关；CD44v6蛋白与MMP-9蛋白两者表达呈显著正相关。 OPN、CD44v6、MMP-9mRNA在47例喉鳞癌组织中的阳性表达率分别为61.7%(29/47)、72.3%(34/47)、66%(31/47)；10例正常喉黏膜组织中的阳性率分别为10%(1/10)、30%(3/10)、0%(0/10),喉鳞癌组织中OPN、CD44v6、MMP-9mRNA的阳性表达率明显高于正常喉黏膜组织中的表达率,差异显著(P0.05)。 OPN、CD44v6mRNA在喉鳞癌组织中高表达与肿瘤分化程度、TNM分期、淋巴结转移相关；OP、CD44v6mRNA与年龄、性别及肿瘤位置等临床病理学特征无明显关系；MMP-9mRNA在喉鳞癌组织中高表达与TNM分期、淋巴结转移相关,MMP-9mRNA与患者年龄、性别、肿瘤位置、分化程度无明显关系。在47例喉鳞癌组织标本中OPN. CD44v6、MMP-9mRNA阳性表达分别为29例、34例、31例,在OPN mRNA阳性表达29例中CD44v6mRNA阳性表达26例,MMP-9mRNA阳性表达24例,经Spearman相关分析,结果显示：OPN mRNA与CD44v6mRNA两者表达呈显著正相关；OPN mRNA与MMP-9mRNA两者表达呈显著正相关；CD44v6mRNA与MMP-9mRNA两者表达呈显著正相关。在47例喉鳞癌组织标本中,OPN蛋白阳性表达为30例,在30例OPN蛋白阳性标本中OPN mRNA阳性29例；CD44v6蛋白阳性表达为36例,在36例CD44v6蛋白阳性标本中CD44v6mRNA阳性34例；MMP-9蛋白阳性表达为32例,在32例MMP-9蛋白阳性标本中MMP-9mRNA阳性31例,经Kappa检验分析,p均小于0.01,说明两种方法具有高度的一致性。 OPN、CD44v6、MMP-9在喉鳞癌组织中的阳性表达率明显高于在正常喉黏膜组织中的阳性表达率,提示OPN、CD44v6、MMP-9可能与喉鳞癌的发生发展有关；OPN、CD44v6、MMP-9在免疫组化和原位杂交两种方法检测中具有高度的一致性；免疫组化及原位杂交均证明OPN、CD44v6、MMP-9在有淋巴结转移的原发性喉鳞癌组织中的表达明显高于无淋巴结转移组,提示OPN、CD44v6、MMP-9与喉鳞癌的转移有关；OPN、CD44v6、MMP-9表达的水平均与肿瘤浸润的范围成正相关,提示OPN与喉鳞癌浸润有关；等级相关分析显示,CD44v6、MMP-9的表达均与OPN有显著相关性,提示三者可能在喉鳞癌的侵袭转移中起协同作用,OPN表达增加可能使得CD44v6和MMP-9表达上调,导致癌细胞的侵袭转移。联合检测OPN、CD44v6和MMP-9对预测肿瘤早期转移和预后有重要意义,OPN将来可能成为喉鳞癌靶向治疗的一个新靶点。
[Abstract]:This paper using immunohistochemical techniques (immunohistochemistry, IHC) and in situ hybridization (In situ hybridization, ISH) detection of laryngeal squamous cell carcinoma (laryngeal squamous cell carcinoma, LSCC) osteopontin tissue and normal laryngeal mucosa (osteopontin, OPN), cell adhesion molecule CD44 (adhesion molecules CD44splice 6 splice variants of variant6, CD44v6), matrix metalloproteinase -9 (matrix metalloproteinase-9 MMP-9) expression of OPN, CD44v6, MMP-9mRNA and protein expression in normal laryngeal mucosa tissues and laryngeal squamous cell carcinoma and relationship with clinical pathological features of patients; to investigate OP, CD44v6, and the relationship between the expression of MMP-9mRNA protein in laryngeal squamous cell carcinoma.
By using immunohistochemistry and in situ hybridization method for the detection of 47 cases of laryngeal squamous cell carcinoma, 10 cases of normal laryngeal mucosa in OPN, CD44v6, MMP-9mRNA and protein expression; three indexes related with age, gender analysis, type, degree of differentiation, the relationship between the expression of the relationship between TNM staging and lymph node metastasis and the three; analysis of two methods for detection of OPN, CD44v6, the consistency of the expression of MMP-9.
OPN, CD44v6, MMP-9 protein expression in 47 cases of laryngeal squamous cell carcinoma were 63.8% (30/47), 76.6% (36/47), 68.1% (32/47); the positive rate of 10 cases of normal laryngeal mucosa tissues were 10% (1/10), 30% (3/10), 0% (0/10), laryngeal squamous cell carcinoma in OPN, CD44v6, the positive expression rate of MMP-9 protein expression was significantly higher than that in normal laryngeal mucosa tissues rate, significant difference (P0.05) of.OPN, CD44v6 protein in laryngeal squamous cell carcinoma with high expression and tumor differentiation, TNM staging, lymph node metastasis, OPN, CD44v6 protein and age, sex and tumor clinical pathology the location features no obvious relationship; MMP-9 protein in laryngeal squamous cell carcinoma with high expression of TNM staging, lymph node metastasis, MMP-9 protein with age, gender, tumor location, no obvious relationship between the degree of differentiation.
In 47 cases of laryngeal squamous cell carcinoma samples OPN, CD44v6, MMP-9 protein expression were 30 cases, 36 cases, 32 cases, the positive expressions of OPN protein and CD44v6 protein in 30 cases of 27 cases, the positive expression of MMP-9 protein in 24 cases, by Spearman analysis, the results showed that OPN protein and CD44v6 protein expression a significant positive correlation; OPN protein and MMP-9 protein expression were positively correlated; the expression of CD44v6 protein was positively correlated with MMP-9 protein of the two.
OPN, CD44v6, MMP-9mRNA positive expression in 47 cases of laryngeal squamous cell carcinoma were 61.7% (29/47), 72.3% (34/47), 66% (31/47); the positive rate of 10 cases of normal laryngeal mucosa tissues were 10% (1/10), 30% (3/10), 0% (0/10), laryngeal squamous cell carcinoma OPN, CD44v6, MMP-9mRNA positive expression rate was significantly higher than that of normal laryngeal mucosa in the rate of significant difference (P0.05). OPN, CD44v6mRNA in laryngeal squamous cell carcinoma with high expression and tumor differentiation, TNM staging, lymph node metastasis; OP, CD44v6mRNA and age, gender and tumor location and clinical pathological characteristics no obvious relationship; MMP-9mRNA in laryngeal squamous cell carcinoma with high expression of TNM staging, lymph node metastasis, MMP-9mRNA and patient age, gender, tumor location, no obvious relationship between the degree of differentiation.
OPN. in 47 cases of laryngeal squamous cell carcinoma samples CD44v6, MMP-9mRNA positive expression were 29 cases, 34 cases, 31 cases, the positive expression of OPN positive expression of mRNA CD44v6mRNA in 29 cases of 26 cases, the positive expression of MMP-9mRNA in 24 cases, by Spearman analysis, the results showed that OPN mRNA and CD44v6mRNA two expression was positively correlated the expression of MMP-9mRNA and OPN; mRNA had a significant positive correlation; the expression of CD44v6mRNA was positively correlated with the MMP-9mRNA of the two.
In 47 cases of laryngeal squamous cell carcinoma tissues, the positive expression of OPN protein in 30 cases, in 30 cases of OPN protein in OPN positive samples of 29 cases were mRNA positive; the positive expression of CD44v6 protein in 36 cases, in 36 cases of CD44v6 positive specimens of 34 cases were CD44v6mRNA positive; the positive expression of MMP-9 protein in 32 cases, 31 cases were MMP-9mRNA positive in 32 cases of MMP-9 positive samples, analyzed by Kappa test, P was less than 0.01, the two methods are highly consistent.
OPN, CD44v6, MMP-9 expression in laryngeal squamous cell carcinoma was significantly higher than the positive expression in normal laryngeal mucosa rate, suggesting that OPN, CD44v6, and MMP-9 may be related to the occurrence and development of laryngeal squamous cell carcinoma; OPN, CD44v6, MMP-9 with a high degree of consistency in immunohistochemistry and in situ hybridization in two methods; immunohistochemistry and in situ hybridization showed that OPN, CD44v6, expression of MMP-9 in lymph node metastasis of primary laryngeal squamous cell carcinoma was significantly higher than that in the group without lymph node metastasis, suggesting that OPN, CD44v6, MMP-9 and metastasis of laryngeal squamous cell carcinoma; OPN, CD44v6, MMP-9 expression levels were associated with tumor invasion a positive correlation, suggesting that OPN and laryngeal squamous cell carcinoma invasion; correlation analysis show that CD44v6, MMP-9 expression was significantly correlated with OPN, suggesting that the three may play a synergistic role in the invasion and metastasis of laryngeal squamous cell carcinoma, the overexpression of OPN may make CD44v6 The increased expression of MMP-9 and the invasion and metastasis of cancer cells will be important. Combined detection of OPN, CD44v6 and MMP-9 is of great significance for predicting the early metastasis and prognosis of cancer. OPN may become a new target for targeted therapy of laryngeal squamous cell carcinoma.

【学位授予单位】：河北北方学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.65

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