宫颈鳞癌转移淋巴结乙酰肝素酶的表达及临床意义

发布时间：2018-02-08 14:42

本文关键词： 宫颈癌乙酰肝素酶 CK19 淋巴结预后　出处：《郑州大学》2014年硕士论文　论文类型：学位论文

【摘要】：背景与目的流行病学调查显示，宫颈癌发病率有上升趋势，严重威胁着女性的生命，位居女性恶性肿瘤死因之首。宫颈癌最主要的转移途径是淋巴转移，而肿瘤的侵润与远处转移是恶性肿瘤治疗成功的主要障碍，淋巴转移是目前明确的导致临床治疗失败和死亡的重要原因。肿瘤的浸润转移能力由肿瘤细胞的多种生物学行为决定，其中肿瘤细胞对其周围基质蛋白的降解能力是一个重要方面，乙酰肝素酶（heparanase，HPA）是一种葡萄糖醛酸内切酶，它能在数个位点降解硫酸乙酰肝素，破坏细胞外基质(extracellular matrix，ECM)和基膜(basement membrane，BM)的完整性，促进肿瘤的局部侵袭和远处转移。我们的前期研究及一些国外的研究已证实HPA在宫颈癌原发灶中的表达显著增高，是宫颈癌的重要生物标志物。细胞角蛋白19（Cytokeratin19，CK19）为细胞角蛋白家族中分子量最小的成员，是构成鳞状上皮细胞骨架的重要蛋白质之一，表达范围严格，特异表达于女性生殖道上皮，已成为上皮性肿瘤细胞较为敏感和特异的标志。国内外都在寻找宫颈癌转移淋巴结特异性和敏感性的生物分子指标，如能在术前准确的预测淋巴结的转移情况，即可准确的了解肿瘤的范围，选择合适的治疗方案，但目前对于淋巴结转移的诊断，尚缺乏有效的手段。基于HPA与宫颈癌以及与淋巴结转移的关系，本研究通过免疫组织化学法检测宫颈鳞癌原发灶及转移淋巴结中HPA的表达情况，首次分析了宫颈鳞癌淋巴结HPA的表达与临床预后因素间的关系，以期为宫颈鳞癌淋巴结的转移诊断提供一新的分子指标。材料与方法收集郑州大学第二附属医院病理科2007年1月-2012年12月确诊的、临床资料完整的宫颈鳞癌石蜡标本102例，其中术后常规病理诊断为淋巴结转移者53例（A组）、淋巴结未转移者49例（B组）。A组选取原发灶、转移淋巴结及未转移淋巴结3个部位，B组选取原发灶和淋巴结2个部位，用于免疫组织化学染色。免疫组织化学S-P法分别检测A、B两组所选部位组织中的HPA表达情况，同时行常规HE染色，HPA阳性的淋巴结行CK19染色，以证实其为转移的宫颈癌病灶。图像分析技术记录各组中HPA表达的平均灰度值，计算出阳性单位。采用SPSS17.0统计软件进行分析，图像分析数据采用x±s表示，各组间比较采用方差分析；HPA表达率采用卡方检验；生存曲线采用Kaplan-Meier方法；生存率比较采用Log-rank检验；多因素分析采用Cox回归分析法。检验水平α=0.05。结果 1.术后常规病理诊断淋巴结转移组（A组）中HPA的表达情况：A组原发灶和转移淋巴结中HPA呈阳性表达均为48例，阳性率为90.6%，术后常规病理诊断未转移淋巴结HPA呈阳性表达7例（13.2%）；原发灶、转移淋巴结与未转移淋巴结中HPA平均灰度值逐渐增高，而阳性单位逐渐降低，组内比较，差异均有统计学意义（F灰度值=30.521，P灰度值=0.000；F阳性单位=6.351，P阳性单位=0.021）； 2.术后常规病理诊断无淋巴结转移组（B组）中HPA的表达情况：B组原发灶HPA呈阳性表达40例（81.6%），术后常规病理诊断未转移淋巴结HPA呈阳性表达5例（10.2%）；B组原发灶HPA平均灰度值明显低于淋巴结（F=21.397，P=0.000），原发灶HPA阳性单位明显高于淋巴结（F=4.827，P=0.037）。 3.宫颈鳞癌中HPA的表达与宫颈鳞癌临床病理特征的关系：淋巴结HPA的表达率与宫颈鳞癌分期（P=0.013）、分化程度（P=0.000）、淋巴结转移（P=0.000）均密切相关。 4.宫颈鳞癌中HPA的表达与生存时间的关系：Kaplan-Meier生存分析显示，所有患者的中位生存时间（MOS，median overall survival）为（49.0±5.9）个月，淋巴结转移者MOS为36.0个月，明显低于淋巴结未转移者的58.5个月（P=0.023）；淋巴结HPA阳性表达者MOS为38.5个月，，淋巴结HPA阴性MOS为57.0个月，二者比较差异有统计学意义（P=0.040）。Cox模型多因素分析显示淋巴结转移、HPA表达是影响宫颈癌患者预后的独立危险因素（P 0.05）。结论 1. HPA在宫颈鳞癌原发灶和转移淋巴结中的表达均显著增高，淋巴结转移是宫颈癌的独立预后因素，可作为宫颈鳞癌淋巴结转移的生物学检测指标； 2.宫颈鳞癌淋巴结转移者MOS明显低于淋巴结未转移者，淋巴结HPA阳性表达者MOS明显低于淋巴结HPA阴性表达者，提示淋巴结HPA阳性患者预后差。
[Abstract]:Background and purpose
Epidemiological investigation showed that the incidence of cervical cancer has a rising trend, a serious threat to women's lives, the first cause of death among the malignant tumors in female. The main route of metastasis of cervical cancer is lymph node metastasis, and tumor invasion and metastasis is a major obstacle to successful treatment of malignant tumors, lymph node metastasis is the important cause of clear death and treatment failure.
The invasion and metastasis ability of tumor is determined by a variety of biological behavior of tumor cells, the degradation ability of tumor cells to the surrounding matrix protein is an important aspect of heparanase (heparanase, HPA) is an endoglycosidase, it can degrade heparan sulfate in a number of sites, disruption of the extracellular matrix (extracellular matrix. ECM (basement) and membrane, BM) membrane integrity, local invasion and distant metastasis. Study on promoting our previous studies and some foreign HPA has been confirmed in primary cervical cancer foci were significantly increased in expression, is an important biomarker of cervical cancer. The expression of cytokeratin 19 (Cytokeratin19, CK19) members of molecular cell keratin family in minimum amount, constitutes one of the important proteins in squamous cell skeleton, the expression scope strictly, specifically expressed in the epithelium of the female genital tract, has become Epithelial tumor cells are more sensitive and specific.
Both at home and abroad are looking for biomarkers of cervical cancer metastasis lymph node specificity and sensitivity, such as accurate in the preoperative prediction of lymph node metastasis, can accurately understand the extent of tumor, choose the appropriate treatment plan, but the diagnosis of lymph node metastasis, the lack of effective means and HPA. Cervical cancer and the relationship with lymph node metastasis based on the expression of this study through the detection of cervical squamous cell carcinoma immunohistochemistry of primary tumors and metastatic lymph nodes in HPA, first analyzes the relationship between the factors and the prognosis of cervical squamous cell carcinoma with lymph node expression between the HPA, in order to provide a new molecular marker for diagnosis of lymph node metastasis node of cervical squamous cell carcinoma.
Materials and methods
From the Second Affiliated Hospital of Zhengzhou University, Department of Pathology, January 2007 -2012 year in December were the complete clinical data of 102 cases of cervical squamous cell carcinoma specimens, including postoperative routine pathological diagnosis for lymph node metastasis in 53 cases (A group), 49 cases without lymph node metastasis (B group).A group selected from the primary tumor, lymph node metastasis and without lymph node metastasis in 3 parts, B group selected primary lesions and lymph nodes in 2 sites, for immunohistochemical staining.
A were detected by immunohistochemical S-P method, B two group of selected parts of the expression of HPA, while conventional HE staining, CK19 staining positive lymph node HPA, to confirm the transfer of cervical cancer lesions. Image analysis techniques were recorded in the expression of HPA in the average gray value, calculate the positive unit.
SPSS17.0 statistical software was used for analysis, image analysis and data representation by X + s, analysis of variance was used to compare between groups; the expression rate of HPA using chi square test method using Kaplan-Meier; survival curve; survival rate compared with Log-rank test; multivariate analysis using Cox regression analysis method. The test level was =0.05.
Result
Pathological diagnosis of lymph node metastasis after operation in group 1. (A group) the expression of HPA in group A: primary tumor and lymph node metastasis in HPA positive expression was found in 48 cases, the positive rate was 90.6%, postoperative routine pathological diagnosis without lymph node metastasis of HPA positive expression in 7 cases (13.2%); original primary tumor, lymph node metastasis and non metastasis lymph nodes in the average gray value of HPA gradually increased, while the positive unit decreased gradually, the comparison group, the differences were statistically significant (F =30.521 gray value of gray value, P =0.000; F =6.351 positive units, P positive =0.021);
2. routine pathological diagnosis after surgery without lymph node metastasis group (B group) the expression of HPA in group B: primary HPA positive expression in 40 cases (81.6%), postoperative routine pathological diagnosis without lymph node metastasis of HPA positive expression in 5 cases (10.2%); B group of primary foci of average gray HPA value was significantly lower than that of lymph node (F=21.397, P=0.000), the primary unit was significantly higher than that of HPA positive lymph nodes (F=4.827, P=0.037).
3., the expression of HPA in cervical squamous cell carcinoma is related to the clinicopathological characteristics of cervical squamous cell carcinoma. The expression rate of HPA in lymph nodes is closely related to the stage of cervical squamous cell carcinoma (P=0.013), the degree of differentiation (P=0.000), and lymph node metastasis (P=0.000).
The relationship between the expression of HPA in 4. cervical squamous cell carcinoma and survival time: Kaplan-Meier survival analysis showed that the median survival time of all patients (MOS median, overall survival) for (49 + 5.9) months, MOS lymph node metastasis was 36 months, 58.5 months was significantly lower than that without lymph node metastasis (P=0.023 lymph node); the positive expression of HPA MOS was 38.5 months, lymph node negative HPA MOS for 57 months, there was a significant difference between the two groups (P=0.040).Cox model multivariate analysis showed that lymph node metastasis, the expression of HPA affect the prognosis of patients with cervical cancer independent risk factor (P 0.05).
conclusion
1. HPA expression was significantly increased in primary squamous cell carcinoma and metastatic lymph nodes. Lymph node metastasis is an independent prognostic factor of cervical cancer. It can be used as a biological indicator for lymph node metastasis of cervical squamous cell carcinoma.
2. cervical squamous cell carcinoma lymph node metastasis MOS was significantly lower than that of lymph node metastasis. Lymph node HPA positive expression of MOS was significantly lower than that of lymph node HPA negative expression, suggesting that lymph node HPA positive patients have poor prognosis.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

【参考文献】