上皮细胞黏附因子调控下咽癌生长转移作用机制的研究

发布时间：2018-05-05 01:19

  本文选题：下咽癌 + EpCAM　； 参考：《山东大学》2014年硕士论文

【摘要】：下咽癌是头颈部常见的恶性肿瘤之一,近年来,尽管手术结合放化疗的综合治疗不断发展,但是下咽癌的5年生存率仍保持在15%-30%,未有明显提高,治疗失败的主要原因是肿瘤的转移及复发。因此,寻找肿瘤转移关键因子并探讨肿瘤增殖转移的潜在机制,对下咽癌的治疗具有重要意义。上皮细胞黏附因子(epithelial cell adhesion molecule, EpCAM)是一种跨膜糖蛋白,在多种肿瘤中高表达,且其表达与肿瘤的增殖转移相关。有关研究证实,EpCAM与头颈部肿瘤的侵袭转移密切相关,但其具体作用存在争议。而关于EpCAM表达对下咽癌转移的影响及机制仍不清楚。本研究检测了EpCAM在下咽癌组织中的表达情况,通过干扰下咽癌FaDu细胞系中EpCAM的表达,探讨了EpCAM因子对下咽癌转移及增殖的影响,并进一步探讨其作用机制。 目的 1.探讨EpCAM在下咽癌组织的表达及其表达强度与T分期、N分期的关系。 2.探讨EpCAM对下咽癌FaDu细胞系侵袭转移和增殖克隆的影响。 3.初步探讨EpCAM影响FaDu细胞侵袭转移增殖克隆的作用机制。 材料与方法 以40例人下咽癌组织及10例正常上皮组织制作石蜡标本,免疫组化法检测EpCAM的表达,以下咽癌FaDu细胞系为实验对象,运用EpCAM siRNA干扰FaDu细胞EpCAM的表达。Transwell检测EpCAM对FaDu细胞系迁移能力和侵袭能力的影响；平板集落形成实验检测EpCAM对FaDu细胞克隆形成能力的影响。细胞增殖实验(MTT法)观察EpCAM对FaDu细胞增殖能力的影响；Western Blot检测干扰EpCAM表达后致瘤和转移相关因子E-cadherin, α-catenin,和P-catenin的表达变化。 结果 1. EpCAM在下咽癌组织中高表达,免疫组化法发现EpCAM主要在下咽癌组织的细胞膜表达,且其表达强度随着下咽癌组织T分期以及N分期的增高而增强。 2.下咽癌FaDu细胞经EpCAM siRNA干扰后,在RNA水平和蛋白质水平EpCAM表达显著降低。 3.降低EpCAM表达后可以抑制FaDu细胞侵袭能力和迁移能力。 4.降低EpCAM表达后可以抑制FaDu细胞增殖能力及克隆能力。 5.降低EpCAM表达后,细胞骨架中的E-cadherin, a-catenin,和β-catenin表达显著升高,而在细胞质中的E-cadherin, a-catenin,和β-catenin表达降低,总蛋白水平无明显变化；细胞核中的β-catenin表达降低。 结论 1. EpCAM在下咽癌组织中高表达,并且其表达水平与肿瘤的T分期,N分期相关。 2. EpCAM通过调节致瘤和转移相关因子E-cadherin, a-catenin,和β-catenin在细胞不同部位的表达变化促进下咽癌FaDu细胞的侵袭、迁移、增殖以及克隆形成能力。 3. EpCAM在下咽癌的恶性行为中发挥重要作用,有可能成为下咽癌治疗的新靶点。
[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, 伪 -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.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.63

【参考文献】

相关期刊论文 前2条

1 郭星,史艳春,费声重,潘子民;下咽癌颈淋巴转移相关因素的研究[J];中华耳鼻咽喉头颈外科杂志;2005年10期

2 侯睿哲;彭云香;毛静涛;侯治富;;胃癌表达EpCAM和CK19临床意义的研究[J];中国实验诊断学;2010年05期

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本文编号：1845542