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標題: | 第二十型黏蛋白在胃癌進展的角色
The role of mucin 20 in the gastric cancer progression |
作者: | Chi-Ju Wu 吳季儒 |
指導教授: | 賴逸儒(I-Rue Lai) |
關鍵字: | 胃癌,黏蛋白,受體鉻胺酸激酶,上皮生長因子接受器,訊息傳遞與轉錄激活酶3, gastric cancer,mucin,receptor tyrosine kinases (RTKs),epidermal growth factor receptor (EGFR),signal transducer and activator of transcription 3 (STAT3), |
出版年 : | 2020 |
學位: | 碩士 |
摘要: | 背景: 胃癌是台灣十大常見癌症之一,即使透過外科手術、化學藥物等方式來治療,胃癌病患在治療之後的五年存活率卻只有20-25%,而如此低的存活率主要與腫瘤的生長速度以及癌細胞的轉移有關。過去研究發現黏蛋白(mucin, MUC)可以當作訊息接受器或者透過細胞膜上的受體鉻胺酸激酶 (receptor tyrosine kinases, RTK)來調控細胞內的訊息傳遞並且藉此影響癌細胞的惡性程度。例如,腫瘤組織若表現大量MUC1的大腸直腸癌病患,其預後較差。研究也發現,MUC20的表現和子宮內膜癌、大腸直腸癌及胰臟癌等惡性程度皆有關係。然而,MUC20的表現和功能是否與胃癌惡性程度相關?目前還尚未得知。 目的: 探討MUC20是否會影響胃癌的惡性程度以及調控胃癌轉移的機制。 實驗材料與方法: 以免疫組織化學染色(immunohistochemical staining)及西方墨點法(western blot)探討胃癌病患的胃腫瘤組織檢體中MUC20的表現和臨床病理參數的相關性。以腸型胃腺癌細胞株AGS探討MUC20 過度表達對於細胞增生能力(crystal violet staining)、爬行能力(transwell migration assay)、侵襲能力(transwell invasion assay)的影響。並且使用人類受體鉻胺酸激酶晶片與西方墨點法分析MUC20過度表達是否會影響胃癌細胞株AGS中受體鉻胺酸激酶 (receptor tyrosine kinases, RTKs)及其下游蛋白的活化。 結果: 根據21個腸型胃腺癌檢體的西方墨點法分析結果,有較多(15/21個)病患其MUC20在胃腫瘤組織中的表現量相較於非腫瘤部分的黏膜組織高。根據71個病患腸型胃腺癌檢體的免疫組織化學染色結果,當胃腫瘤組織中MUC20表現較高(score>7)的時候,其癌細胞分化較差的比率(11/26 v.s 6/37)以及且發生遠端轉移(7/26 v.s 1/37)的機會均較MUC20表現低的那組病患高。在細胞實驗, MUC20過度表達的AGS細胞株會有較強的爬行能力及侵襲能力。相較於控制組(mock)細胞株,過度表達MUC20的胃癌細胞株其上皮生長因子接受器(Epidermal growth factor receptor, EGFR)的磷酸化程度較高,且EGFR的下游訊息傳遞與轉錄激活酶3(Signal transducer and activator of transcription3, STAT3)也會被活化。 結論: 腸型胃腺癌組織比非腫瘤部分的黏膜組織表現出較高MUC20。表現出高MUC20的胃癌病患,有較高機會出現遠處器官轉移且其癌細胞分化較差。從細胞實驗發現, MUC20過度表達的胃癌細胞株具有較強的爬行能力以及侵襲能力,這個表現型可能和MUC20活化EGFR路徑的機制有關。 Background: Gastric cancer (GC) is one of the ten most common cancers in Taiwan. Even though some of the GC patients were cured by surgical resection and/or chemotherapy, the overall 5-year survival rate of GC was only twenty to twenty-five percent, mostly due to the uncontrolled cancer growth and metastasis. Previous studies reported that mucin glycoproteins could function as receptors or bind to receptor tyrosine kinases (RTKs) to regulate signal transduction and cancer progression. For instance, the colorectal cancer patients with mucin 1 (MUC1) overexpression had poorer prognosis than those without. Several studies also demonstrated that mucin 20 (MUC20) was associated with endometrial, colorectal and pancreatic adenocarcinoma. However, the expression level and function of MUC20 in GC is still unclear. Aims: To investigate whether MUC20 could affect gastric cancer progression and the regulatory mechanism of MUC20 in gastric cancer. Materials and methods: Western blot analysis was performed to study the expression of MUC20 in the tumorous and non-tumorous tissue of the intestinal type gastric adenocarcinomas. Immunohistochemical staining was used to analyzed the correlation of the MUC20 expression in the cancerous part with clinico-pathological parameters of patients with the intestinal type gastric adenocarcinomas. AGS cells were transfected with MUC20/pcDNA3.1A vector to overexpress MUC20. Cell viability, migration, and invasion were analyzed by crystal violet staining, transwell migration and matrigel invasion assays, respectively. Human phospho-receptor tyrosine kinase array and western blot analysis were used to investigate the effects of MUC20 overexpression on RTK signaling in AGS cells. Results: Fifteen of the twenty-one samples assessed by western blot analysis showed MUC20 overexpression in the cancerous part of intestinal type gastric adenocarcinoma, when compared to the the non-tumorous mucosa tissue. The immunohistochemical staining of 71 gastric cancer tissue samples showed that higher percentage of poor cell differentiation (11/26 v.s 6/37) and distant metastasis (7/26 v.s 1/37) in MUC20 high expression (scores>7) group than MUC20 low expression (scores<6) group. In vitro studies showed that overexpression of MUC20 in AGS cells enhanced their abilities to migrate and invade. Moreover, MUC20 overexpression increased phosphorylation of epidermal growth factor receptor (EGFR) and signal transducer and activator of transcription 3 (STAT3) in AGS cells. Conclusions: Our findings suggested that MUC20 was overexpressed in the intestinal type gastric adenocarcinoma tissue compared with the non-tumorous mucosa tissue. The patients with MUC20 high expression had higher chances of distant metastasis, and their cancer cells often were classified as poorly-differentiated. Our in vitro studies suggested that MUC20 may increase gastric cancer cell migration and invasion through EGFR pathway. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76745 |
DOI: | 10.6342/NTU202004002 |
全文授權: | 未授權 |
顯示於系所單位: | 解剖學暨細胞生物學科所 |
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