本文選題：下咽癌 + EpCAM　； 參考：《山東大學》2014年碩士論文

【摘要】：下咽癌是頭頸部常見的惡性腫瘤之一,近年來,盡管手術結合放化療的綜合治療不斷發(fā)展,但是下咽癌的5年生存率仍保持在15%-30%,未有明顯提高,治療失敗的主要原因是腫瘤的轉移及復發(fā)。因此,尋找腫瘤轉移關鍵因子并探討腫瘤增殖轉移的潛在機制,對下咽癌的治療具有重要意義。上皮細胞黏附因子(epithelial cell adhesion molecule, EpCAM)是一種跨膜糖蛋白,在多種腫瘤中高表達,且其表達與腫瘤的增殖轉移相關。有關研究證實,EpCAM與頭頸部腫瘤的侵襲轉移密切相關,但其具體作用存在爭議。而關于EpCAM表達對下咽癌轉移的影響及機制仍不清楚。本研究檢測了EpCAM在下咽癌組織中的表達情況,通過干擾下咽癌FaDu細胞系中EpCAM的表達,探討了EpCAM因子對下咽癌轉移及增殖的影響,并進一步探討其作用機制。 目的 1.探討EpCAM在下咽癌組織的表達及其表達強度與T分期、N分期的關系。 2.探討EpCAM對下咽癌FaDu細胞系侵襲轉移和增殖克隆的影響。 3.初步探討EpCAM影響FaDu細胞侵襲轉移增殖克隆的作用機制。 材料與方法 以40例人下咽癌組織及10例正常上皮組織制作石蠟標本,免疫組化法檢測EpCAM的表達,以下咽癌FaDu細胞系為實驗對象,運用EpCAM siRNA干擾FaDu細胞EpCAM的表達。Transwell檢測EpCAM對FaDu細胞系遷移能力和侵襲能力的影響；平板集落形成實驗檢測EpCAM對FaDu細胞克隆形成能力的影響。細胞增殖實驗(MTT法)觀察EpCAM對FaDu細胞增殖能力的影響；Western Blot檢測干擾EpCAM表達后致瘤和轉移相關因子E-cadherin, α-catenin,和P-catenin的表達變化。 結果 1. EpCAM在下咽癌組織中高表達,免疫組化法發(fā)現(xiàn)EpCAM主要在下咽癌組織的細胞膜表達,且其表達強度隨著下咽癌組織T分期以及N分期的增高而增強。 2.下咽癌FaDu細胞經EpCAM siRNA干擾后,在RNA水平和蛋白質水平EpCAM表達顯著降低。 3.降低EpCAM表達后可以抑制FaDu細胞侵襲能力和遷移能力。 4.降低EpCAM表達后可以抑制FaDu細胞增殖能力及克隆能力。 5.降低EpCAM表達后,細胞骨架中的E-cadherin, a-catenin,和β-catenin表達顯著升高,而在細胞質中的E-cadherin, a-catenin,和β-catenin表達降低,總蛋白水平無明顯變化；細胞核中的β-catenin表達降低。 結論 1. EpCAM在下咽癌組織中高表達,并且其表達水平與腫瘤的T分期,N分期相關。 2. EpCAM通過調節(jié)致瘤和轉移相關因子E-cadherin, a-catenin,和β-catenin在細胞不同部位的表達變化促進下咽癌FaDu細胞的侵襲、遷移、增殖以及克隆形成能力。 3. EpCAM在下咽癌的惡性行為中發(fā)揮重要作用,有可能成為下咽癌治療的新靶點。
[Abstract]:Hypopharyngeal carcinoma is one of the most common malignant tumors in the head and neck. In recent years, despite the continuous development of surgery combined with radiotherapy and chemotherapy, the 5-year survival rate of hypopharyngeal carcinoma has remained at 15-30% and has not improved significantly. The main cause of failure is metastasis and recurrence of tumor. Therefore, it is important for the treatment of hypopharyngeal carcinoma to search for the key factors of tumor metastasis and to explore the potential mechanism of tumor proliferation and metastasis. Epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein, which is highly expressed in many kinds of tumors, and its expression is related to the proliferation and metastasis of tumor. It has been confirmed that EpCAM is closely related to the invasion and metastasis of head and neck tumors, but its role is controversial. However, the effect of EpCAM expression on hypopharyngeal carcinoma metastasis and its mechanism are still unclear. In this study, we examined the expression of EpCAM in hypopharyngeal carcinoma. By interfering with the expression of EpCAM in hypopharyngeal carcinoma FaDu cell line, we investigated the effect of EpCAM factor on metastasis and proliferation of hypopharyngeal carcinoma, and further explored its mechanism. Purpose 1. To investigate the expression of EpCAM in hypopharyngeal carcinoma and its relationship with T staging and N stage. 2. To investigate the effect of EpCAM on invasion, metastasis and proliferation of hypopharyngeal carcinoma FaDu cell line. 3. To explore the mechanism of EpCAM affecting the invasion and metastasis of FaDu cells. Materials and methods Paraffin wax specimens were prepared from 40 cases of human hypopharyngeal carcinoma and 10 cases of normal epithelium. The expression of EpCAM was detected by immunohistochemical method. The FaDu cell line of hypopharyngeal carcinoma was used as experimental object. The expression of EpCAM in FaDu cells was interfered by EpCAM siRNA. Transwell was used to detect the effect of EpCAM on the migration and invasion of FaDu cell lines, and the effect of EpCAM on the clone formation of FaDu cells was detected by plate colony forming assay. The effect of EpCAM on the proliferation of FaDu cells was observed. The expression of E-cadherin, 偽 -catenin, and P-catenin were detected by Western Blot after interfering with the expression of EpCAM. Result 1. High expression of EpCAM was found in hypopharyngeal carcinoma tissue. Immunohistochemical method showed that EpCAM was mainly expressed on the cell membrane of hypopharyngeal carcinoma, and its expression intensity was increased with the increase of T stage and N stage of hypopharyngeal carcinoma. 2. The expression of EpCAM in hypopharyngeal carcinoma FaDu cells was significantly decreased at the RNA level and protein level after interference by EpCAM siRNA. 3. Decreasing the expression of EpCAM could inhibit the invasion and migration of FaDu cells. 4. Decreased EpCAM expression could inhibit the proliferation and clone ability of FaDu cells. 5. The expression of E-cadherin, a-catenin, and 尾 -catenin in cytoskeleton increased significantly after EpCAM was decreased, but the expression of E-cadherin, a-catenin, and 尾 -catenin in cytoplasm decreased, but the total protein level did not change, but the expression of 尾 -catenin in nucleus decreased. Conclusion 1. EpCAM was highly expressed in hypopharyngeal carcinoma and its expression level was correlated with T staging and N stage of the tumor. 2. EpCAM promotes the invasion, migration, proliferation and clone formation of hypopharyngeal carcinoma FaDu cells by regulating the expression of E-cadherin, a-catenin, and 尾 -catenin in different parts of the cells. 3. EpCAM plays an important role in the malignant behavior of hypopharyngeal carcinoma and may become a new target for the treatment of hypopharyngeal carcinoma.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R739.63

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本文編號：1845542