本文關(guān)鍵詞： 宮頸癌 乙酰肝素酶 CK19 淋巴結(jié) 預(yù)后　出處：《鄭州大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：背景與目的 流行病學(xué)調(diào)查顯示，宮頸癌發(fā)病率有上升趨勢，嚴(yán)重威脅著女性的生命，位居女性惡性腫瘤死因之首。宮頸癌最主要的轉(zhuǎn)移途徑是淋巴轉(zhuǎn)移，而腫瘤的侵潤與遠(yuǎn)處轉(zhuǎn)移是惡性腫瘤治療成功的主要障礙，淋巴轉(zhuǎn)移是目前明確的導(dǎo)致臨床治療失敗和死亡的重要原因。 腫瘤的浸潤轉(zhuǎn)移能力由腫瘤細(xì)胞的多種生物學(xué)行為決定，其中腫瘤細(xì)胞對其周圍基質(zhì)蛋白的降解能力是一個重要方面，乙酰肝素酶（heparanase，HPA）是一種葡萄糖醛酸內(nèi)切酶，它能在數(shù)個位點降解硫酸乙酰肝素，破壞細(xì)胞外基質(zhì)(extracellular matrix，ECM)和基膜(basement membrane，BM)的完整性，促進腫瘤的局部侵襲和遠(yuǎn)處轉(zhuǎn)移。我們的前期研究及一些國外的研究已證實HPA在宮頸癌原發(fā)灶中的表達(dá)顯著增高，是宮頸癌的重要生物標(biāo)志物。細(xì)胞角蛋白19（Cytokeratin19，CK19）為細(xì)胞角蛋白家族中分子量最小的成員，是構(gòu)成鱗狀上皮細(xì)胞骨架的重要蛋白質(zhì)之一，表達(dá)范圍嚴(yán)格，特異表達(dá)于女性生殖道上皮，已成為上皮性腫瘤細(xì)胞較為敏感和特異的標(biāo)志。 國內(nèi)外都在尋找宮頸癌轉(zhuǎn)移淋巴結(jié)特異性和敏感性的生物分子指標(biāo)，如能在術(shù)前準(zhǔn)確的預(yù)測淋巴結(jié)的轉(zhuǎn)移情況，即可準(zhǔn)確的了解腫瘤的范圍，選擇合適的治療方案，但目前對于淋巴結(jié)轉(zhuǎn)移的診斷，尚缺乏有效的手段�；贖PA與宮頸癌以及與淋巴結(jié)轉(zhuǎn)移的關(guān)系，本研究通過免疫組織化學(xué)法檢測宮頸鱗癌原發(fā)灶及轉(zhuǎn)移淋巴結(jié)中HPA的表達(dá)情況，首次分析了宮頸鱗癌淋巴結(jié)HPA的表達(dá)與臨床預(yù)后因素間的關(guān)系，以期為宮頸鱗癌淋巴結(jié)的轉(zhuǎn)移診斷提供一新的分子指標(biāo)。 材料與方法 收集鄭州大學(xué)第二附屬醫(yī)院病理科2007年1月-2012年12月確診的、臨床資料完整的宮頸鱗癌石蠟標(biāo)本102例，其中術(shù)后常規(guī)病理診斷為淋巴結(jié)轉(zhuǎn)移者53例（A組）、淋巴結(jié)未轉(zhuǎn)移者49例（B組）。A組選取原發(fā)灶、轉(zhuǎn)移淋巴結(jié)及未轉(zhuǎn)移淋巴結(jié)3個部位，B組選取原發(fā)灶和淋巴結(jié)2個部位，用于免疫組織化學(xué)染色。 免疫組織化學(xué)S-P法分別檢測A、B兩組所選部位組織中的HPA表達(dá)情況，同時行常規(guī)HE染色，HPA陽性的淋巴結(jié)行CK19染色，以證實其為轉(zhuǎn)移的宮頸癌病灶。圖像分析技術(shù)記錄各組中HPA表達(dá)的平均灰度值，計算出陽性單位。 采用SPSS17.0統(tǒng)計軟件進行分析，圖像分析數(shù)據(jù)采用x±s表示，各組間比較采用方差分析；HPA表達(dá)率采用卡方檢驗；生存曲線采用Kaplan-Meier方法；生存率比較采用Log-rank檢驗；多因素分析采用Cox回歸分析法。檢驗水平α=0.05。 結(jié)果 1.術(shù)后常規(guī)病理診斷淋巴結(jié)轉(zhuǎn)移組（A組）中HPA的表達(dá)情況：A組原發(fā)灶和轉(zhuǎn)移淋巴結(jié)中HPA呈陽性表達(dá)均為48例，陽性率為90.6%，術(shù)后常規(guī)病理診斷未轉(zhuǎn)移淋巴結(jié)HPA呈陽性表達(dá)7例（13.2%）；原發(fā)灶、轉(zhuǎn)移淋巴結(jié)與未轉(zhuǎn)移淋巴結(jié)中HPA平均灰度值逐漸增高，而陽性單位逐漸降低，組內(nèi)比較，差異均有統(tǒng)計學(xué)意義（F灰度值=30.521，P灰度值=0.000；F陽性單位=6.351，P陽性單位=0.021）； 2.術(shù)后常規(guī)病理診斷無淋巴結(jié)轉(zhuǎn)移組（B組）中HPA的表達(dá)情況：B組原發(fā)灶HPA呈陽性表達(dá)40例（81.6%），術(shù)后常規(guī)病理診斷未轉(zhuǎn)移淋巴結(jié)HPA呈陽性表達(dá)5例（10.2%）；B組原發(fā)灶HPA平均灰度值明顯低于淋巴結(jié)（F=21.397，P=0.000），原發(fā)灶HPA陽性單位明顯高于淋巴結(jié)（F=4.827，P=0.037）。 3.宮頸鱗癌中HPA的表達(dá)與宮頸鱗癌臨床病理特征的關(guān)系：淋巴結(jié)HPA的表達(dá)率與宮頸鱗癌分期（P=0.013）、分化程度（P=0.000）、淋巴結(jié)轉(zhuǎn)移（P=0.000）均密切相關(guān)。 4.宮頸鱗癌中HPA的表達(dá)與生存時間的關(guān)系：Kaplan-Meier生存分析顯示，所有患者的中位生存時間（MOS，median overall survival）為（49.0±5.9）個月，淋巴結(jié)轉(zhuǎn)移者MOS為36.0個月，明顯低于淋巴結(jié)未轉(zhuǎn)移者的58.5個月（P=0.023）；淋巴結(jié)HPA陽性表達(dá)者MOS為38.5個月，，淋巴結(jié)HPA陰性MOS為57.0個月，二者比較差異有統(tǒng)計學(xué)意義（P=0.040）。Cox模型多因素分析顯示淋巴結(jié)轉(zhuǎn)移、HPA表達(dá)是影響宮頸癌患者預(yù)后的獨立危險因素（P 0.05）。 結(jié)論 1. HPA在宮頸鱗癌原發(fā)灶和轉(zhuǎn)移淋巴結(jié)中的表達(dá)均顯著增高，淋巴結(jié)轉(zhuǎn)移是宮頸癌的獨立預(yù)后因素，可作為宮頸鱗癌淋巴結(jié)轉(zhuǎn)移的生物學(xué)檢測指標(biāo)； 2.宮頸鱗癌淋巴結(jié)轉(zhuǎn)移者MOS明顯低于淋巴結(jié)未轉(zhuǎn)移者，淋巴結(jié)HPA陽性表達(dá)者MOS明顯低于淋巴結(jié)HPA陰性表達(dá)者，提示淋巴結(jié)HPA陽性患者預(yù)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.

【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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