PRL-3、MMP-14、CD105在人喉鳞状细胞癌中的表达及意义

发布时间：2018-06-17 11:51

本文选题：肝再生磷酸酶-3 + 基质金属蛋白酶-14　；参考：《郑州大学》2011年硕士论文

【摘要】：喉鳞状细胞癌是头颈部最常见的恶性肿瘤之一,95%-98%的喉部恶性肿瘤为鳞状细胞癌,目前其主要的治疗手段是以根治性手术辅以放疗或精确的放疗为主,在早期患者可以获得比较好的局部控制率,但不能预防或减少远处转移,对于中晚期患者常规治疗的疗效不尽人意,大概局部复发率27-61%、远处转移率18-21%、五年生存率27-34%,随着肿瘤生物学的发展许多生物学治疗的新技术运用到临床,实际运用中许多现象和结果至今都无法解释。头颈部肿瘤的浸润和转移是影响患者生存率的主要原因,但肿瘤由原发灶向周边组织浸润以致发生远处转移的机制目前尚未清楚,癌细胞是如何获得浸润能力的尚不清楚!探索新的、特异性高的肿瘤标志物,对提高喉癌患者的预后和进一步明确喉癌由原发灶向周边组织浸润以致发生远处转移的机制有重要意义。肝再生磷酸酶-3(PhosPhatase of regenerating liver-3, PRL-3)是新近发现的蛋白酪氨酸磷酸酶,其通过蛋白去磷酸化实现调节蛋白功能,参与各种细胞的生命活动,研究表明,PRL-3通过促进细胞增殖、迁移、侵袭、粘附及血管生成等途径参与了多种肿瘤的发生,浸润及转移。基质金属蛋白酶-14(Matrix metailo Proteinase-14, MMP-14)属膜型基质金属蛋白酶类,存在于细胞膜上,是一种跨膜蛋白,具有降解细胞外基质的能力,这是恶性肿瘤形成的必须条件之一。 CD105又名endoglin,是一种糖蛋白,CD105阳性染色主要定位于新生血管内皮细胞的细胞膜和细胞质,在大血管内皮细胞中不表达,是理想的血管内皮细胞标记物,是计数肿瘤微血管密度(MVD)的金标准即CD105-MVD。目前,国内外关于PRL-3的研究主要集中在腺癌方面,其在鳞癌方面的研究尚少,在喉鳞癌方面的研究国内外还未见报道。PRL-3参与肿瘤浸润和转移的机制尚不清楚,有文献报道PRL-3可能通过基质金属蛋白酶和血管生成途径参与肿瘤的浸润和转移,但PRL-3、MMP-14及CD105在喉鳞癌中的表达及相关性尚未见报道。本课题应用HE染色和免疫组化法对人喉鳞癌中的PRL-3、MMP-14蛋白的表达进行检测,并用CD105标记肿瘤组织的微血管密度(CD105-MVD),旨在探索其与人喉癌的发生、浸润和转移的关系,寻找可能预测喉癌浸润和转移的肿瘤标志物,为喉鳞癌患者的预后提供有益的观测指标,并为进一步的机制研究提供理论依据。目的检测PRL-3、MMP-14在人喉鳞癌、相应癌旁组织和正常喉黏膜组织中的表达,并用CD105标记微血管密度(CD105-MVD),探讨PRL-3、MMP-14和CD105-MVD在喉鳞状细胞癌各临床病理因素中的作用及相关性,旨在探索其与人喉癌的发生、浸润及转移的关系,寻找可能预测喉癌浸润和转移的肿瘤标志物,为喉鳞癌患者的预后提供有益的观测指标,并为进一步的机制研究提供理论依据。材料与方法 1、研究对象选择郑州大学第一附属医院2006年1月～2008年12月收治的69例喉鳞状细胞癌患者的癌组织蜡块标本,其中高分化鳞状细胞癌28例,中-低分化鳞状细胞癌41例(低分化癌病例较少,与中分化病例合为中-低分化组)。伴淋巴结转移阳性者25例,淋巴结转移阴性者44例。按国际抗癌协会(UICC) TNM分类标准(2002)分期：Ⅰ期11例,Ⅱ期15例,Ⅲ期27例,Ⅳ期16例。病人年龄在40岁至84岁之间(平均62岁),男67例,女2例,所有患者临床资料完整,术前均未作放疗、化疗及任何免疫治疗。另取35例同批患者的癌旁组织,32例正常喉黏膜组织作对照,癌旁组织取自肉眼观察距肿瘤边缘1.0 cm以外的喉黏膜组织,均经病理证实为鳞状上皮不典型增生,未发现癌细胞。 2、实验方法采用免疫组织化学方法检测69例人喉鳞状细胞癌组织、35例相应癌旁组织和32例正常喉黏膜组织中PRL-3和MMP-14的表达情况,并用CD105作为血管内皮标记物对喉癌组织的MVD进行标记,结合临床病理因素分析。 3、统计学处理采用SPSS18.0软件包进行统计分析,样本率的比较采用x检验,计量资料用x±s表示,均数比较采用t验,PRL-3、MMP-14与CD105-MVD相关性采用Spearman等级相关分析,以a=0.05为检验水准。结果 1、PRL-3在喉鳞癌组织中的阳性表达率(65.22%)高于癌旁组织(17.14%)和正常喉黏膜组织(15.63%),差异有统计学意义(P0.01)。淋巴结转移组的PRL-3阳性表达率明显高于无淋巴结转移组,差异具有统计学意义(P0.05)。PRL-3的表达与病理分期、组织分化程度及淋巴结转移相关,与肿瘤原发部位、性别及年龄无关。 2、MMP-14在喉鳞癌组织中的阳性表达率(76.81%)高于癌旁组织(31.43%)和正常喉黏膜组织(25.00%),差异显著,有统计学意义(P0.01)。淋巴结转移阳性组的MMP-14蛋白阳性表达率明显高于淋巴结转移阴性组,差异显著,具有统计学意义(P0.05)。MMP-14的表达与病理分期、组织分化程度和淋巴结转移相关,与肿瘤原发部位、性别及年龄无关。 3、临床病理分期较晚的Ⅲ～Ⅳ的CD105-MVD (12.7907±4.16080)明显高于Ⅰ～Ⅱ期(9.8846±4.13112),淋巴结转移阳性组的CD105-MVD(13.4000±4.36845)高于无淋巴结转移组(10.7273±4.08835),差异均有统计学意义(P0.05或P0.01)。CD105-MVD值与分化程度、原发部位、年龄及性别无关,比较差异无统计学意义(P0.05)。 4、PRL-3表达与CD105-MVD呈正相关(γs=0.811,P0.01)；MMP-14表达与CD105-MVD呈正相关(r=0.867,P0.01)； 5、PRL-3蛋白与MMP-14蛋白的阳性表达率差异无统计学意义(P0.05),但PRL-3蛋白的表达与MMP-14的表达呈正相关(r=0.367,P0.01)。 1、PRL-3、MMP-14参与了喉鳞状细胞癌的发生、浸润及转移,对喉鳞癌的诊断有重要参考价值； 2、PRL-3与MMP-14的表达呈正相关,二者可能促进喉癌的浸润和转移,可联合作为喉鳞癌标志物,对临床治疗方案的选择及评估预后有一定的参考价值； 3、CD105是理想的肿瘤新生血管标记物,CD105-MVD可作为预测喉癌转移及评估预后的重要指标； 4、PRL-3、MMP-14与肿瘤血管的形成密切相关,在喉鳞状细胞癌形成中起重要作用。
[Abstract]:Laryngeal squamous cell carcinoma (laryngeal squamous cell carcinoma) is one of the most common malignant tumors in the head and neck. The malignant tumor of the larynx of 95%-98% is squamous cell carcinoma. At present, the main means of treatment are radical surgery supplemented by radiotherapy or precise radiotherapy, and a better local control rate can be obtained in early patients, but it can not prevent or reduce distant metastasis. The curative effect of conventional treatment in advanced patients is not satisfactory, the local recurrence rate is 27-61%, the distant metastasis rate is 18-21%, the five year survival rate is 27-34%. With the development of tumor biology, many new techniques of biological treatment are applied to the clinic. Many phenomena and results are not explained so far in practical application.
The invasion and metastasis of the head and neck tumors are the main factors that affect the survival rate of the patients. However, the mechanism of tumor invasion from the primary tissue to the peripheral tissue and the distant metastasis is not yet clear. It is not clear how the cancer cells can obtain the infiltration capacity. Explore new, high specific tumor markers to improve the prognosis and advance of the patients with larynx cancer. One step is to clarify the mechanism of laryngeal cancer infiltrating from the primary to the surrounding tissues, leading to distant metastasis.
The liver regenerative phosphatase -3 (PhosPhatase of regenerating liver-3, PRL-3) is a newly discovered protein tyrosine phosphatase, which regulates protein function through protein dephosphorylation and participates in the life activities of various cells. The study shows that PRL-3 has been involved in many swells by promoting cell proliferation, migration, invasion, adhesion and angiogenesis. The occurrence, infiltration and metastasis of the tumor.
Matrix metalloproteinase -14 (Matrix metailo Proteinase-14 (MMP-14)) is a membrane type matrix metalloproteinase, which exists on the cell membrane. It is a transmembrane protein and has the ability to degrade the extracellular matrix, which is one of the necessary conditions for the formation of malignant tumors.
CD105, also known as endoglin, is a kind of glycoprotein. CD105 positive staining is mainly located in the cell membrane and cytoplasm of the neovascular endothelial cells. It is not expressed in the large vascular endothelial cells. It is an ideal vascular endothelial cell marker. It is the golden standard for counting the tumor microvascular density (MVD), that is, CD105-MVD.
At present, domestic and foreign research on PRL-3 mainly focuses on adenocarcinoma, and its research on squamous cell carcinoma is few. There is no report on the mechanism of.PRL-3 involvement in tumor invasion and metastasis at home and abroad. It is reported that PRL-3 may be involved in tumor infiltration through matrix metalloproteinase and angiogenesis pathway. The expression and correlation of PRL-3, MMP-14 and CD105 in laryngeal squamous cell carcinoma have not yet been reported. The purpose of this study was to detect the expression of PRL-3, MMP-14 protein in human laryngeal squamous cell carcinoma by HE staining and immunohistochemical method, and to mark the microvascular density (CD105-MVD) of the tumor tissue with CD105 in order to explore the occurrence, infiltration and metastasis of human larynx cancer. The relationship is to find the tumor markers that may predict the invasion and metastasis of laryngeal carcinoma, providing useful observations for the prognosis of the patients with laryngeal squamous cell carcinoma and providing a theoretical basis for further mechanism research.
objective
The expression of PRL-3 and MMP-14 in human laryngeal squamous cell carcinoma, corresponding paracancerous tissue and normal laryngeal mucosa, and CD105 labeled microvascular density (CD105-MVD) were used to explore the role and correlation of PRL-3, MMP-14 and CD105-MVD in the various clinicopathological factors of laryngeal squamous cell carcinoma, in order to explore the relationship with the occurrence, infiltration and metastasis of human larynx cancer. The tumor markers that may predict the invasion and metastasis of laryngeal carcinoma provide useful observations for the prognosis of the patients with laryngeal squamous cell carcinoma and provide a theoretical basis for further mechanism research.
Materials and methods
1, the object of research
69 cases of squamous cell carcinoma of the larynx were selected from the First Affiliated Hospital of Zhengzhou University from January 2006 to December 2008, including 28 cases of highly differentiated squamous cell carcinoma, 41 cases of medium low differentiated squamous cell carcinoma (low differentiated carcinoma cases, middle and low differentiation with middle differentiation cases). 25 cases with lymph node metastasis were found. According to the international anticancer association (UICC) TNM classification standard (UICC) classification standard (2002) stage: 11 cases, 15 cases in stage II, 27 cases in stage III, 16 cases in stage IV. The patients were aged from 40 to 84 years (average 62 years old), 67 men and 2 cases, all patients were complete with clinical data, no preoperative radiotherapy, chemotherapy and any immunotherapy. And 35 cases of the same batch In the para cancerous tissue, 32 normal laryngeal mucosa tissues were compared. The para cancerous tissue was taken from the naked eye to observe the laryngeal mucosa outside the tumor margin 1 cm from the tumor. All of them were proved to be atypical hyperplasia of squamous epithelium by pathology, and no cancer cells were found.
2, experimental method
The expression of PRL-3 and MMP-14 in 69 cases of human laryngeal squamous cell carcinoma (laryngeal squamous cell carcinoma), 35 cases of corresponding para cancerous tissue and 32 normal laryngeal mucosa tissues were detected by immunohistochemistry. The MVD of laryngeal carcinoma tissue was labeled with CD105 as vascular endothelial marker, combined with clinicopathological analysis.
3, statistical treatment
The SPSS18.0 software package was used for statistical analysis. The comparison of sample rate was compared with X test. The measurement data was expressed in X + s. The average number was compared to t, PRL-3, MMP-14 and CD105-MVD, and a=0.05 was used as the test level.
Result
1, the positive expression rate of PRL-3 in laryngeal squamous cell carcinoma (65.22%) was higher than that of paracancerous tissue (17.14%) and normal laryngeal mucosa (15.63%). The difference was statistically significant (P0.01). The positive rate of PRL-3 in the lymph node metastasis group was significantly higher than that in the non lymph node metastasis group, and the difference was statistically significant (P0.05).PRL-3 expression and pathological stage, tissue differentiation. The degree and lymph node metastasis were not related to the primary location, sex and age of the tumor.
2, the positive expression rate of MMP-14 in the laryngeal squamous cell carcinoma (76.81%) was higher than that of the paracancerous tissue (31.43%) and normal laryngeal mucosa (25%). The difference was significant (P0.01). The positive rate of MMP-14 protein in the lymph node metastasis positive group was significantly higher than that of the lymph node metastasis negative group, and the difference was significant (P0.05).MMP-14 table It was related to pathological stage, histological differentiation and lymph node metastasis, and was not related to the primary location, sex and age of the tumor.
3, CD105-MVD (12.7907 + 4.16080) of late stage of clinicopathological stage (12.7907 + 4.16080) was significantly higher than that of stage I to II (9.8846 + 4.13112), CD105-MVD (13.4000 + 4.36845) of lymph node metastasis positive group was higher than that of no lymph node metastasis group (10.7273 + 4.08835). The difference was statistically significant (P0.05 or P0.01).CD105-MVD value and differentiation degree, primary site, year Age and sex were not related, the difference was not statistically significant (P0.05).
4, PRL-3 expression was positively correlated with CD105-MVD (gamma s=0.811, P0.01); MMP-14 expression was positively correlated with CD105-MVD (r=0.867, P0.01).
5, there was no significant difference in the positive expression rate of PRL-3 protein and MMP-14 protein (P0.05), but the expression of PRL-3 protein was positively correlated with the expression of MMP-14 (r=0.367, P0.01).
1, PRL-3 and MMP-14 participate in the occurrence, invasion and metastasis of laryngeal squamous cell carcinoma, and have important reference value for the diagnosis of laryngeal squamous cell carcinoma.
2, the expression of PRL-3 is positively correlated with the expression of MMP-14. The two may promote the invasion and metastasis of larynx cancer, and can be combined as a marker of laryngeal squamous cell carcinoma, which has a certain reference value for the selection of clinical treatment scheme and evaluation of prognosis.
3, CD105 is an ideal marker for tumor angiogenesis. CD105-MVD can be used as an important indicator for predicting metastasis and evaluating prognosis of laryngeal carcinoma.
4, PRL-3 and MMP-14 are closely related to tumor angiogenesis and play an important role in the formation of laryngeal squamous cell carcinoma.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.65

【参考文献】