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Title: | NKX2.5 基因在鼻咽癌之功能分析 Functional analysis of NKX2.5 gene in Nasopharyngeal Carcinoma |
Authors: | Bing-Sheng-Lu 呂秉昇 |
Advisor: | 林欽塘(Chin-Tarng Lin) |
Keyword: | 鼻咽癌,腫瘤發生,NKX2.5 之體內外功能, Nasopharyngeal carcinoma (NPC),NPC progression,NKX2.5, |
Publication Year : | 2018 |
Degree: | 碩士 |
Abstract: | 鼻咽癌是一種好發於東南亞,尤其是中國南部、香港、台灣的頭頸部癌並且與許多致病因素有關。環境因素:如廣東高鹽醃製食物以及EBV病毒的感染都是被證實的。鼻咽癌的致病機轉如同其他實質固態瘤一樣目前還是難以解釋。本論文研究目的,就是想找出與鼻咽癌癌化過程中有明顯改變表達的基因。在之前先使用了cDNA微陣列分析(cDNA microarray analysis)的方法,比較鼻咽癌細胞與正常鼻咽黏膜上皮細胞兩者之間的差異,我們發現轉錄因子NKX2.5 基因在鼻咽癌細胞中有較高的表現量。接下來為了探討NKX2.5 是否能夠影響鼻咽癌的癌化現象,首先分析NKX2.5在不同的鼻咽癌細胞株的表現,發現在NPC-TW-01細胞株中比正常鼻黏膜上皮細胞有較高的NKX2.5表達。此外我們透過免疫組織染色以及西方墨點法發現NKX2.5蛋白在NPC-TW-01細胞株與病人檢體中表現量均增加。為了進一步分析NKX2.5的功能在鼻咽癌細胞中所扮演的角色,利用NKX2.5的short hairpin RNA (shRNA) 來抑制NKX2.5的基因表現。在抑制NKX2.5基因表現後可以發現鼻咽癌細胞的增生、轉移以及侵襲能力皆有顯著的降低。接下來在異種移植鼻咽癌腫瘤(NPC xenografts)於免疫缺陷小鼠的實驗中顯示:以抑制NKX2.5 基因表現的shRNA所轉染的鼻咽癌細胞,腫瘤生長速度會有減緩。我們的發現也說明了NKX2.5可能在鼻咽癌癌化過程中扮演一個促進的角色,同時也說明NKX2.5 是鼻咽癌致病機轉中也有所影響包括癌細胞侵襲、增生、轉移。這個新的發現NKX2.5 在鼻咽癌所扮演的重要功能對於此癌症之分子標靶治療及有相當的潛力。 Nasopharyngeal carcinoma (NPC) is one of common head and neck cancers in Southeast Asia, particularly Southern China, Hong Kong and Taiwan, believed to have a multifactorial etiology. Environmental factors, consumption of Cantonese salted food and infections of Epstein-Barr virus (EBV) are most documented. The pathogenesis of NPC, like that of most solid tumors, remains elusive. The aim of this study was to identify the significant genes that may be altered during NPC progression. Previously, using cDNA microarray analysis, and we found that the transcription factor NKX2.5. NKX2.5 showed a significant different expression and highly expressed in NPC cell lines than normal nasal mucosal epithelial cells. In order to investigate that whether NKX2.5 gene could affect NPC progression, we observed at first the gene expression in different NPC cells lines using qRT-PCR analysis and found that expression of NKX2.5 in NPC cell lines, especially in NPC-TW01 cell line was highly upregulated. We also demonstrate that the expression level of NKX2.5 protein was increased in NPC-TW01 NPC cell line and NPC biopsy specimens by immunohistochemical staining and western bloting. In order to analysis of the functional role of NKX2.5 in NPC-TW01 cell line, the expression of NKX2.5 was significantly reduced that compared with the control in NPC-TW01 after knockdown of NKX2.5 by NKX2.5 shRNA-lentiviral infection. This in turn resulted in a significant reduction of cell proliferation, invasion, and migration activity of NPC cells. However, in NOD/SCID mice bearing NKX2.5 transfected NPC xenograft, the tumor growth was significantly decreased. Our findings strongly suggest that NKX2.5 may play a role to promote the formation of nasopharyngeal carcinoma of nasopharynx and as a gene in NPC pathogenesis to affect NPC migration, proliferation and invasion. These novel findings of functional role of NKX2.5 may have a potential implication in molecular targeted therapy for NPC. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71236 |
DOI: | 10.6342/NTU201801832 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 病理學科所 |
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